Molecular docking and in vivo studies of Aloe vera gel for their potential anti-apoptotic activity against aluminium chloride-induced neurotoxicity in mice

Molecular docking and in vivo studies of Aloe vera gel for their potential anti-apoptotic activity against aluminium chloride-induced neurotoxicity in mice | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Molecular docking and in vivo studies of Aloe vera gel for their potential anti-apoptotic activity against aluminium chloride-induced neurotoxicity in mice Azubuike Raphael Nwaji, Ruth C. Nwosu, Chibuzor S. Amadi, Esther O. Ngwu, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7332209/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Neurotoxicity resulting from aluminium chloride (AlCl₃) exposure has been linked to increased apoptosis in neuronal cells, contributing to various neurodegenerative diseases. On the other hand, the use of alternative medicines has been practiced for years and formed part of our tradition around the world. This study investigated the impact of Aloe vera gel on brain apoptotic and pro-apoptotic markers in mice subjected to aluminium chloride-induced neurotoxicity via experimental and computational models. Twenty mice were divided into four groups as control, AlCl₃, Aloe vera and AlCl₃ + Aloe vera respectively. Following a two-week treatment period, brain tissues were harvested for quantitative determination of apoptotic markers, employing standard methods. Molecular docking was done on all identified ligands and proteins using a computational approach. The results demonstrated a significant increase in Caspase-3 and Bax levels in the AlCl₃ group compared to both the control and Aloe vera groups. Conversely, Bcl-2 levels were significantly reduced in the AlCl₃ group. Notably, treatment with Aloe vera gel led to decreased Casp-3 and Bax levels in the AlCl₃ + Aloe vera group compared to the AlCl₃ group, while Bcl-2 levels increased. The molecular docking analysis of the present study indicated that Aloe resin C had the best binding score for all the proteins sampled with scores of -8.180, -6.908 and − 9.811 for Bax, Bcl-2 and Casp-3 respectively. These findings thus, highlight the potentials of Aloe vera as a therapeutic agent in stabilizing neuronal membrane and mitigating apoptotic processes associated with aluminium chloride exposure by promoting anti-apoptotic signaling. Biological sciences/Biochemistry Biological sciences/Chemical biology Biological sciences/Drug discovery Biological sciences/Neuroscience Docking Aluminium chloride Neurotoxicity Apoptosis Aloe vera Figures Figure 1 Introduction Neurotoxicity, a term that refers to the condition in which the nervous system is directly or indirectly exposed to chemicals either naturally occurring or man-made, thereby impairing the nervous system’s normal functioning in both man and animals [ 1 ], [ 2 ]. As reported by [ 3 ], [ 4 ], the brain is an important organ that is more susceptible to exogenous stimuli than other organs of the body because of numerous polyunsaturated fatty acid contents which makes the neural tissues very sensitive to peroxidation. Several chemicals can elicit neurotoxic disruptions in man, and also many are employed in experimental models for neurotoxicity in animals, such as aluminium chloride. Aluminium chloride is an abundant metal on earth both in the environment and in food, with easy access to the human body via different means [ 5 ], [ 6 ]. It is a well-established neurotoxicant involved in the etiology of neurodegenerative diseases [ 7 ]. For instance, it has been reported by several researchers that it is a major neurotoxin, disrupter of neurological functions and accelerator of oxidative stress, plaque formation and amyloid beta (Aβ) deposition in animal models, humans and cell lines [ 8 ]–[ 11 ]. According to [ 11 ], apoptosis is the final step in the pathway of toxicity induced by aluminium leading to neurodegeneration. Apoptosis is a controlled form of physiological cell death involving cascades of degenerative events [ 12 ], [ 13 ]. Accordingly, experimental and clinical evidence have established that neurological insults, including chemical exposures, can trigger aberrant apoptosis in the brain in vitro or in vivo [ 7 ], [ 13 ], [ 14 ]. For instance, various apoptotic markers (Bcl-2 and caspase families) have been reported to be expressed following aluminium exposure indicating programmed cell death [ 15 ]. The specific mechanisms of apoptosis induction vary between exposure paradigms. However, apoptosis is triggered generally by increasing pro-apoptotic and/or decreasing anti-apoptotic markers. Alternative medicine usage has been in practiced for years as still being used today, even in a more advanced way, including Aloe vera. According to [ 16 ], alternative medicine remains a constituent part of many cultural and technological development around the world. Aloe vera also referred to as Aloe Barbadensis Miller , belonging to the Liliaceae family is a plant with long-standing reputation majorly for its health, skincare and other benefits [ 17 ]. Research has shown that Aloe vera contains several phytochemicals and about 200 bioactive components, with the inner gel reported to possess most of the components [ 17 ]. The plant has been reported to possess various pharmacological properties, including antioxidant, antidiabetic, wound healing, anticancer, immune system regulations and memory-enhancing properties [ 18 ]–[ 21 ]. These potentials were attributed to its bioactive components such as phenolic, anthraquinone, flavonoids, terpenoids and saponins [ 16 ]. Thus, emphasizing its potential as a valuable natural therapeutic agent. Despite all the numerous biological potentials attributed to Aloe vera, no docking-in vivo work has reported on the possible role of Aloe vera in ameliorating apoptosis induced by aluminium chloride in the brain. Following the recent enhancements in search algorithms and energy function, computational designing of a therapeutic agent and molecular docking strategies has become valuable tools in exploring the interaction between proteins and ligands, coupled with inexpensive and streamlined techniques aiding potential therapeutic targets discoveries in specific pathophysiological situations [ 22 ], [ 23 ]. This study therefore, aims to investigate the impact of Aloe vera gel on brain apoptotic and proapoptotic markers of aluminium induced neurotoxicity-mice and explore the biological mechanisms through molecular docking studies. Materials and methods In vivo study Chemical Material Aluminium Chloride (AlCl₃) was obtained from Sigma-Aldrich (Sigma-Aldrich Cooperation, USA). Every other reagent utilized was of high quality and analytical grade. The reporting of this study as well conforms to ARRIVE 2.0 guidelines [24]. Ethical statement Authors declare that the study protocol was approved by the Ethics and Animal Handling Committee of the Faculty of Basic Medical Sciences, Alex Ekwueme Federal University, Ndufu-Alike, Nigeria (approval number: AE-FUNAI/FBMS/EAHC/25/012). Plant Material and extraction Leaves of Aloe vera plant were obtained from Ikwo in Ebonyi State. The leaves were identified and authenticated by Mr. C.B. Udechukwu, a botanist in the Department of Biological Sciences, assigned a voucher specimen number, AE-FUNAI/H/1220 and was deposited in the AE-FUNAI Herbarium. Extraction was done by selecting the matured Aloe vera leaves from the base of the plant. The leaves were cleaned and washed of dusts. Then, the thick green skin on both sides was removed thereby exposing the clear gel. The fresh gel was collected by scraping and squeezing into a container. The extraction was freshly done each time of the administration. Experimental Animals Twenty male Swiss mice (weighing 20-30 grams) were procured from University of Uyo, Akwa Ibom State. The mice were acclimatized for three weeks prior to the study and housed in plastic cages. The animals were exposed to 12-hour light and 12-hour dark cycles. The study was conducted with the animals cared for and humanely treated in accordance with the Guide for the Care and Use of Laboratory Animals. Experimental Design The mice were randomly allocated to four groups of five rats each (n=5) as follows: Control Group: Mice received no treatment (0.5 mL normal saline) Aluminium Chloride-Induced Toxicity Group: Mice received aluminium chloride (100 mg/kg body weight, orally) throughout the experimental period Aloe vera Group: Mice were treated with Aloe vera gel (200 mg/kg body weight) daily throughout the experimental period via oral gavage Aluminium Chloride + Aloe vera Group: Mice were exposed to aluminium chloride (100mg/kg) and treated with Aloe vera (200 mg/kg). The treatment protocols lasted for two weeks. Sample collection On the last day of experimental period, the mice were humanely sacrificed and euthanized by employing ketamine/xylazine (0.1 mL/100 g.bw, intraperitoneally). Following euthanasia, their brains were removed and put in 0.25M cold sucrose buffer solution and were later homogenized for biochemical assays. B iochemical assessment Estimation of caspase-3, Bcl-2 and Bax were done quantitatively by standard methods following the manufacturer’s guidelines using Caspase Assay Kit; G-Bioscience), (BCL Assay Kit; G-Bioscience) and Bax assay Kit respectively. Results were expressed as μmol/mg protein. In silico study Preparation of Target Protein and Ligands In the present study, the forcefield employed for ligand preparation, protein preparation, and molecular docking was OPLS4. The protein databank (PDB) (http://www.pdb.org) was used to retrieve the crystal structures of proteins, viz., Bax (PDB ID: 5W62), Bcl-2 (PDB ID: P10417) and Casp-3 (PDB ID: P70677). Preparation of the proteins was carried out at a physiological pH range of 7.0 ± 0.4. The structure of the ligands of Aloe vera which included the Arachidonic acid (PubChem CID: 444899), Beta-carotene (PubChem CID: 5280489), 3-Epi-beta-sitosterol (PubChem CID: 12303645), Aloe-emodin PubChem CID: 10207), Campesterol (PubChem CID: 173183), Aloe resin C (PubChem CID: 11972360), Cholesterol (PubChem CID: 5997), Quercetin PubChem CID: 5280343), and Aloin (PubChem CID: 14989), were retrieved from the PubChem database from NCBI (www.pubchem.ncbi.nlm.nih.gov/). Preparation and protonation of ligands were done at a physiological pH of 7.2. Also, the retrieved structures of the ligands were minimized for their potential interactions with the target protein. Molecular docking Molecular docking analysis employed extra precision docking (XP) which was used for the retrieved ligands with their corresponding target proteins (BAX, BCL2, CASP3) retrieved from the protein data bank (PDB). The 3D crystal structure of the mouse ligands was complexed with an inhibitor molecule which was eliminated and docking with the ligands was performed. The proteins were energetically minimized and active sites were predicted with the selection of maximized grid parameters. This id followed by calculation of binding energies for the docked complexes. Higher negative binding energy indicates higher binding affinity. Statistical Analysis The statistical analysis was performed with the help of GraphPad Prism software (GraphPad Software, version 8.0 Inc., San Diego, USA). The results were expressed as Mean ± Standard Error of Mean (SEM). Analysis of data was done using a one-way analysis of variance (ANOVA) and t-test, utilizing Tukey’s post hoc test for comparison of means at 95% (p<0.05) confidence level. Results The results of all the experiments are presented in Tables 1-3 and Figure 1 (a-l) below. Effect of Aloe vera on brain apoptotic and proapoptotic markers in mice As shown in table 1 below, there was a significant increase in Caspase-3 levels in the AlCl₃ group compared to both the Control group (p < 0.05) and the Aloe vera group (p < 0.01). However, treatment with Aloe vera significantly decreased it (p < 0.05). Also, Bax levels were significantly increased in the AlCl₃ group compared to the Aloe vera group and control (p < 0.01). However, there was a significant decline in the Bax levels following treatment with Aloe vera. Meanwhile, Bcl-2 levels significantly decreased in the AlCl₃ group when compared to the Control and Aloe vera groups (p < 0.05), but was significantly alone (p < 0.05) elevated after Aloe vera treatment. In the docking study, Aloe ligands (Aloe resin C, Aloin, Aloe emodin, arachidonic acid and Quercetin) showed favorable binding affinities against Bax, Bcl-2 and Casp-3 as shown in table 3 and figure 1 (a-l) below. Table 1. Effect of Aluminium Chloride and Aloe vera on Brain Apoptotic and Proapoptotic Markers in Mice Parameters Control AlCl 3 Aloe vera AlCl 3 + Aloe vera Casp-3 (µmol/mg) 22.57±1.29 38.70±0.93 ac* 17.60±1.51 33.23±1.48 abc Bax (µmol/mg) 172.70±19.38 364.30±10.27 a*c* 130.00±3.22 263.00±17.10 abc Bcl-2 (µmol/mg) 7.17±0.44 3.33±0.55 ac 12.10±2.23 5.30±0.42 bc (a p<0.05vs control, b p<0.05 vs AlCl 3 , c p<0.05 vs Aloe vera , c* p<0.01 vs Aloe vera; n=5, AlCl 3 = aluminium chloride, Casp-3 = Caspase-3, Bax =Bcl-2 associated X , Bcl-2 =B-cell lymphoma 2 ) Table 2. Phytochemicals of Aloe vera ligand molecules Molecule_ID Name PubChem_CID Smiles MOL001439 Arachidonic acid 444899 CCCCC/C=C\C/C=C\C/C=C\C/C=C\CCCC(=O)O MOL002773 Beta-carotene 5280489 CC1=C(C(CCC1)(C)C)/C=C/C(=C/C=C/C(=C/C=C/C=C(/C=C/C=C(/C=C/C2=C(CCCC2(C)C)C)\C)\C)/C)/C MOL000359 3-Epi-beta-sitosterol 12303645 CC[C@H](CC[C@@H](C)[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2CC=C4[C@@]3(CC[C@H](C4)O)C)C)C(C)C MOL000471 Aloe-emodin 10207 C1=CC2=C(C(=C1)O)C(=O)C3=C(C2=O)C=C(C=C3O)CO MOL005043 Campesterol 173183 C[C@H](CC[C@@H](C)C(C)C)[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2CC=C4[C@@]3(CC[C@@H](C4)O)C)C MOL005051 Aloe resin C 11972360 CC1=CC(=C(C2=C1C(=O)C=C(O2)CC(=O)C)[C@H]3[C@@H]([C@H]([C@@H]([C@H](O3)CO)O)O)OC(=O)/C=C/C4=CC=C(C=C4)O)O[C@H]5[C@@H]([C@H]([C@@H]([C@H](O5)CO)O)O)O MOL000953 Cholesterol 5997 C[C@H](CCCC(C)C)[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2CC=C4[C@@]3(CC[C@@H](C4)O)C)C MOL000098 Quercetin 5280343 C1=CC(=C(C=C1C2=C(C(=O)C3=C(C=C(C=C3O2)O)O)O)O)O MOL005060 Aloin 14989 C1=CC2=C(C(=C1)O)C(=O)C3=C([C@@H]2[C@H]4[C@@H]([C@H]([C@@H]([C@H](O4)CO)O)O)O)C=C(C=C3O)CO Table 3. Binding energies and interaction details of ligands-proteins complexes. Target proteins Ligand PubChem_ CID Docking Score Hydrogen Bond Glide energy BAX (mouse) Aloe resin C 11972360 -8.18 -4.76 -47.071 Quercetin 5280343 -7.626 -3.019 -36.329 Aloin 14989 -4.32 -2.128 -33.171 Aloe-emodin 10207 -3.613 -1.39 -24.227 Cholesterol 5997 -2.22 -0.35 -22.836 3-Epi-beta-sitosterol 12303645 -2.166 0 -20.759 Campesterol 173183 -2.145 -0.35 -20.834 Bcl-2 (mouse) Aloe resin C 11972360 -6.908 -2.534 -48.76 Aloe-emodin 10207 -6.496 -1.44 -35.466 Aloin 14989 -6.296 -1.92 -40.55 Quercetin 5280343 -6.24 -2.4 -33.105 Campesterol 173183 -5.011 -0.239 -19.288 Arachidonic acid 444899 -4.681 -0.35 -29.683 Cholesterol 5997 -4.366 0 -20.848 3-Epi-beta-sitosterol 12303645 -3.879 -0.7 -24.55 Beta-carotene 5280489 -2.67 0 -37.395 Casp-3 (mouse) Aloe resin C 11972360 -9.811 -6.074 -53.796 Aloin 14989 -7.068 -2.971 -40.587 Quercetin 5280343 -6.988 -2.718 -44.023 Arachidonic acid 444899 -5.738 -2.35 -35.585 Aloe-emodin 10207 -5.271 -1.84 -30.506 Cholesterol 5997 -4.432 -0.514 -26.583 3-Epi-beta-sitosterol 12303645 -3.916 -1.187 -24.196 Campesterol 173183 -2.599 -0.7 -22.002 Beta-carotene 5280489 -0.693 0 -38.753 Discussion In vivo study Neurotoxicity exposure has been linked to increased apoptosis in neuronal cells, contributing to various neurodegenerative diseases. For instance, various apoptotic markers have been reported to be expressed following aluminium exposure, indicating programmed cell death. [11] also reported that apoptosis is the final pathway of toxicity induced by aluminium thereby leading to neurodegeneration. Apoptosis plays an important roles in development and tissue homeostasis, and its altered regulation is implicated in degenerative diseases such as Alzheimers’ [25]. Data from our present study reveals elevated concentration of pro-apoptotic proteins- caspase-3 and Bax in the brain tissues of aluminium chloride–induced AlCl 3 - induced neurotoxic rats with decrease of the anti-apoptotic protein Bcl-2, an indication of enhanced apoptotic signaling and reduced cellular protection against apoptosis following AlCl 3 exposure. Conversely, these results also showed that treatment of neurotoxicant rats with Aloe vera gel reversed the altered Bcl-2 and Bax levels induced by aluminium chloride thereby restoring the balance between regulating proteins of apoptosis. These findings align with those of previous studies. For instance, a study by [10], reported an aluminium chloride-induced apoptosis within the brain of Wistar rats through the activation of apoptotic caspase-3. Aluminium has been suggested to accelerate deposition of amyloid beta, plaque formation and oxidative stress in rats brain [10]. Similarly, earlier reports by [26] noted that intracisternal infusion of aluminium into the brain of rabbit decreased Bcl-2 and decreased Bax and caspase-3 levels. This also agrees with the reports of [27] in the hippocampal tissues. Increased activation of caspase as reported by [28] confirmed that Aluminium dose-dependently induces apoptosis in brain of rats. Treatment with Aloe vera however, decreased the pro-apoptotic markers and increased the anti-apoptotic as seen in table 1. This also agrees with the study of [4] who reported that Aloe vera gel ameliorated apoptosis together with oxidative stress and hippocampal neuronal loss, in a study aimed at evaluating the protective effects of Aloe vera gel on cisplatin-induced apoptosis in the hippocampus of rats. Thus, resulting in the down-regulation of Casp-3 and Bax, and up-regulation of Bcl-2. Earlier reports indicated that aloe-emodin, one of the active components of Aloe vera could suppress NMDA- induced apoptosis of retinal ganglion cells, thus suggesting its possible role in glaucoma management [29]. Also, recent reports highlighted the capability of aloe sterols to counteracting apoptosis [30], [31]. However, [17] reported the efficacy of Aloe vera in inducing apoptosis in cancer cells and exhibiting immunomodulatory potentials. According to [32], they appear to initiate apoptosis via up-regulation and down-regulation of the expression of P53 and Bcl-2 gene respectively. Another report by [33], also suggested that an extract of Aloe vera exerts anticancer effects on human breast and cervical cancer cells by inhibiting cancer cell growth and inducing apoptosis. The fate of cell is determined by the measure of balance between pro-apoptotic and anti-apoptotic proteins [34]. When death is favoured by the balance, anti-apoptotic- Bcl-2 inhibits cytochrome c release within the mitochondrial membrane while Bax monitors apoptotic processes by controlling cytochrome c release into the cytoplasm thereby resulting to activation of casp-3 cascade that cleave downstream substrates leading to apoptosis [35], [36]. Chemicals typically cause apoptosis by enhancing pro-apoptotic and/or decreasing anti-apoptotic signaling resulting from oxidative stress, elevated intracellular Ca 2+ levels, or Zn 2+ dyshomeostasis, albeit the precise mechanisms of apoptosis induction differ depending on the exposure paradigm. Molecular docking According to [37], computational screening of natural products have become a necessity due to its easier and cheaper and directional approach. The study involved molecular docking studies of several Aloe vera ligands on proteins involved in apoptosis (Casp-3, Bax and Bcl-2), to assess the binding affinity in terms of binding energy and interaction. In the present study, nine active ligand molecules derived from Aloe vera were performed against the apoptotic proteins. However, five top ligands (aloe emodin, aloin, quercetin, aloe resin C and arachidonic acid; see figure 1) were selected based on their binding affinity and patterns indicated by higher negative docking scores. However, aloe resin C exhibited highest level of interactions with Casp-3, Bax and Bcl-2 active sites. The Bcl-2 family of proteins are key regulators of intracellular apoptotic signal transduction that negate damaging effects of reactive oxygen species and the key biochemical event that defines apoptosis is the activation of caspases [15], [38], [39]. Results of the molecular docking showed that aloe resin C exhibited a H-bonding interactions with Casp3 active sites of GLY 122, ASP 253, SER 205, ARG 207, SER, 63 and SER 120; Bax active sites of LYS 21, ASP 53, GLN 18, PRO 13 and TRP 158; and Bcl2 active site of ALA 146. From molecular docking results, it can be concluded that aloe resin C, aloe emodin, aloin, quercetin, and arachidonic acid interfere with the functions of Casp3, Bax and Bcl2 through H-bonding and hydrophobic interactions, which might suppress the neurodegenerative disease progression. Thereby, portraying the potentials of Aloe vera as a therapeutic agent in enhancing cellular protection and stabilizing neuronal membrane against apoptosis. Conclusion In silico molecular docking study of active ingredients of Aloe vera with apoptotic proteins together with higher binding energy with all biomarker proteins gave a strong hypothesis to its anti-apopototic potentials. Thus, the findings of this study demonstrated the potentials of Aloe vera as a therapeutic agent in enhancing cellular protection and stabilizing neuronal membrane against apoptotic processes associated with aluminium chloride exposure by enhancing and promoting anti-apoptotic signaling. Declarations Funding Statement: This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors. Author contributions: Study conception and design was done by ARN. Material preparation, data collection and analysis were performed by all the authors (ARN, RCN, CSA, EON, AOO, IAA and AEO). The first draft of the manuscript was written and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript. Acknowledgements: The authors are grateful to Asma’u Bala and Adeshina Odugbemi from the School of Pharmacy, University of Western Cape, South Africa for assisting us with the docking studies. Data availability statement: The authors declare that the data supporting the findings of the study are available within the article Declaration of Competing Interest: Authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article. References Bilge, S. Neurotoxicity, Types, Clinical Manifestations, Diagnosis and Treatment. Neurotox. - New. Adv. 1–13. 10.5772/intechopen.101737 (2022). Spencer, P. S. & Lein, P. J. Neurotoxicity, in Encyclopedia of Toxicology: Third Edition , pp. 489–500. (2014). 10.1016/B978-0-12-386454-3.00169-X Song, B., Zhou, T., Liu, J. & Shao, L. Q. Involvement of Programmed Cell Death in Neurotoxicity of Metallic Nanoparticles: Recent Advances and Future Perspectives. Nanoscale Res. Lett. 11 (484), 1–12. 10.1186/s11671-016-1704-2 (2016). Majd, N. E., Shahraki, R., Tabandeh, M. R. & Hosseinifar, S. Protective effects of Aloe vera gel on cisplatin-induced oxidative stress, apoptosis and neurons structure in rat hippocampus. Vet. Res. Forum . 13 (1), 111–119. 10.30466/vrf.2020.119876.2835 (2022). Clement, L., W, J. T., Ordukenneth, S., D, O. C. & Dirokweni, E. Evaluation of Apoptotic Changes Using BCL-2 on Aluminium Chloride Induced Neurotoxicity Treated with Ethanolic Extract of Carpolobia lutea at Different Doses. Asian J. Med. Princ Clin. Pract. 7 (2), 443–460 (2024). Yokel, R. A. & McNamara, P. J. Aluminium Toxicokinetics: An Updated MiniReview. Pharmacol. Toxicol. 88 (4), 159–167. 10.1111/j.1600-0773.2001.880401.x (2001). Skalny, A. V. et al. Molecular mechanisms of aluminum neurotoxicity: Update on adverse effects and therapeutic strategies 1st edn, Vol. 5 (Elsevier Inc., 2021). 10.1016/bs.ant.2020.12.001 Bondy, S. C. NeuroToxicology The neurotoxicity of environmental aluminum is still an issue. Neurotoxicology 31 (5), 575–581. 10.1016/j.neuro.2010.05.009 (2010). Blaylock, R. L. Aluminum Induced Immunoexcitotoxicity in Neurodevelopmental and Neurodegenerative Disorders. Curr. Inorg. Chem. 2 (1), 46–53 (2012). Mesole, S. B., Alfred, O. O., Yusuf, U. A., Lukubi, L. & Ndhlovu, D. Apoptotic Inducement of Neuronal Cells by Aluminium Chloride and the Neuroprotective Effect of Eugenol in Wistar Rats, Oxid. Med. Cell. Longev. , vol. pp. 1–7, 2020, (2020). 10.1155/2020/8425643 Shahani, L. Aluminium induced Neurotoxicity: A Review. Pharmacol. Life Sci. Bull. Env Pharmacol. Life Sci. 9 (3), 63–71 (2020). Dikranian, K. et al. Apoptosis in the in Vivo Mammalian Forebrain. Neurobiol. Dis. 8 , 359–379. 10.1006/nbdi.2001.0411 (2001). Martin, L. J. Neuronal cell death in nervous system development, disease, and injury (Review). Int. J. Mol. Med. 7 , 455–478 (2001). Sastry, P. S. & Rao, K. S. Apoptosis and the Nervous System. J. Neurochem . 74 , 1–20 (2000). Yuan, J. & Yankner, B. A. Apoptosis in the nervous system, Nature , vol. 407, no. October, pp. 802–809, (2000). Ranjith, D. Molecular docking studies of aloe vera for their potential antibacterial activity using Argus lab 4.0. 1. Pharma Innov. J. 8 (9), 481–487 (2019). Kaur, S. & Bains, K. Aloe Barbadensis Miller (Aloe Vera) Pharmacological activities and clinical evidence for disease prevention. Int. J. Vitam. Nutr. Res. 94 , 3–4. 10.1024/0300-9831/a000797 (2024). Drudi, D. et al. Aloe barbadensis Miller versus silver sulfadiazine creams for wound healing by secondary intention in dogs and cats: a randomized controlled study. Res. Vet. Sci. 1–40. 10.1016/j.rvsc.2017.10.010 (2017). Salah, F. et al. Effect of the deacetylation degree on the antibacterial and antibiofilm activity of acemannan from Aloe vera. Ind. Crop Prod. 103 , 13–18. 10.1016/j.indcrop.2017.03.031 (2017). Fox, L. T. et al. In vitro wound healing and cytotoxic activity of the gel and whole-leaf materials from selected aloe species. J. Ethnopharmacol. 1–27. 10.1016/j.jep.2017.02.017 (2017). Redza-dutordoir, M. & Averill-bates, D. A. Activation of apoptosis signalling pathways by reactive oxygen species. BBA - Mol. Cell. Res. 10.1016/j.bbamcr.2016.09.012 (2016). Luty, B. A., Wasserman, Z. R., Stouten, P. F. W. & Hodge, C. N. A Molecular Mechanics / Grid Method for Evaluation of Ligand-Receptor Interactions. J. Comput. Chem. 16 (4), 454–464 (1995). Chavda, V., Patel, V. & Patel, S. Investigation of neuroprotective potential of anti-diabetic agents in ischemic brain proteomics through in-silico molecular simulation studies. Biointerface Res. Appl. Chem. 12 (4), 5347–5362. 10.33263/BRIAC124.53475362 (2022). Percie Du, N. et al. The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. BMJ Open. Sci. 4 (1), 1–7. 10.1136/bmjos-2020-100115 (2020). Strasser, A., Connor, L. O. & Dixit, V. M. Poptosis ignaling. Annu. Rev. Biochem. 69 , 219–245 (2000). Ghribi, O., Herman, M. M., Spaulding, N. K. & Savory, J. Lithium inhibits aluminum-induced apoptosis in rabbit hippocampus, by preventing cytochrome c translocation, Bcl-2 decrease, Bax elevation and caspase-3 activation. J. Neurochem . 82 , 137–145 (2002). Niu, Q. & Zhang, H. Relationship between apoptosis of rat hippocampus cells induced by aluminum and the copy of the bcl-2 as well as bax mRNA and the copy of the bcl 2 as well as bax mRNA. J. Hyg. Res. 34 (6), 671–675 (2005). Zhang, H., Yang, X., Qin, X. & Niu, Q. Caspase-3 is Involved in Aluminum-Induced Impairment of Long- Term Potentiation in Rats Through the Akt / GSK-3 b Pathway. Neurotox. Res. 10.1007/s12640-016-9597-5 (2016). Lin, H. et al. Aloe-emodin Suppressed NMDA-induced Apoptosis of Retinal Ganglion Cells through Regulation of ERK Phosphorylation. Phyther Res. 21 , 1007–1014. 10.1002/ptr (2007). Saito, M. et al. Oral administration of Aloe vera gel powder prevents UVB-induced decrease in skin elasticity via suppression of overexpression of MMPs in hairless mice. Biosci. Biotechnol. Biochem. 7 , 1416–1424. 10.1080/09168451.2016.1156480 (2016). Misawa, E. et al. Protective effects of Aloe sterols against UVB-induced photoaging in hairless mice. Photodermatol Photoimmunol Photomed. 2 , 101–111. 10.1111/ijlh.12426 (2017). Shalabi, M. et al. Asian Pacific Journal of Tropical Biomedicine. Asian Pac. J. Trop. Biomed. 5 (5), 375–381. 10.1016/S2221-1691(15)30372-5 (2015). Hussain, A., Sharma, C., Khan, S., Shah, K. & Haque, S. Aloe vera Inhibits Proliferation of Human Breast and Cervical Cancer Cells and Acts Synergistically with Cisplatin. Asian Pac. J. Cancer Prev. 16 (7), 2939–2946 (2015). Bouillet, P. et al. Degenerative Disorders Caused by Bcl-2 Deficiency Prevented by Loss of Its BH3-Only Antagonist Bim. Dev. Cell. 1 , 645–653 (2001). Cai, X., Zhang, H., Tong, D., Tan, Z. & Han, D. Corosolic Acid Triggers Mitochondria and Caspase - dependent Apoptotic Cell Death in Osteosarcoma MG ‐ 63 Cells. Phyther Res. 25 , 1354–1361 (2011). Magiera, M. M. et al. Trim17-mediated ubiquitination and degradation of Mcl-1 initiate apoptosis in neurons. Cell. Death Differ. 1–12. 10.1038/cdd.2012.124 (2012). Rifaioglu, A. S. et al. Recent applications of deep learning and machine intelligence on in silico drug discovery: methods, tools and databases, Brief. Bioinform. , no. August, pp. 1–35, (2018). 10.1093/bib/bby061 Chipuk, J. E., Moldoveanu, T., Llambi, F., Parsons, M. J. & Green, D. R. Review The BCL-2 Family Reunion. Mol. Cell. 37 (3), 299–310. 10.1016/j.molcel.2010.01.025 (2010). Akifusa, S. et al. Free Radical Biology & Medicine Globular adiponectin-induced RAW 264 apoptosis is regulated by a reactive oxygen species-dependent pathway involving Bcl-2. Free Radic Biol. Med. 46 (9), 1308–1316. 10.1016/j.freeradbiomed.2009.02.014 (2009). Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7332209","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":508937974,"identity":"47c5db55-e3ae-4de1-93a0-b09f3a292ac5","order_by":0,"name":"Azubuike Raphael Nwaji","email":"data:image/png;base64,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","orcid":"","institution":"Alex Ekwueme Federal University","correspondingAuthor":true,"prefix":"","firstName":"Azubuike","middleName":"Raphael","lastName":"Nwaji","suffix":""},{"id":508937975,"identity":"09b1bc7f-659b-422e-a7f8-f1b3b9040c52","order_by":1,"name":"Ruth C. Nwosu","email":"","orcid":"","institution":"Alex Ekwueme Federal University","correspondingAuthor":false,"prefix":"","firstName":"Ruth","middleName":"C.","lastName":"Nwosu","suffix":""},{"id":508937976,"identity":"4d18d8e6-4208-4a80-aeae-b930f950bf0b","order_by":2,"name":"Chibuzor S. Amadi","email":"","orcid":"","institution":"Western Illinois University","correspondingAuthor":false,"prefix":"","firstName":"Chibuzor","middleName":"S.","lastName":"Amadi","suffix":""},{"id":508937977,"identity":"b8c52fde-8353-41bf-8a6d-f8bcf436067b","order_by":3,"name":"Esther O. Ngwu","email":"","orcid":"","institution":"David Umahi Federal University of Health sciences","correspondingAuthor":false,"prefix":"","firstName":"Esther","middleName":"O.","lastName":"Ngwu","suffix":""},{"id":508937978,"identity":"46182157-ecfb-469c-ad46-e1449a2cf8e9","order_by":4,"name":"Anyigor O. Ogah","email":"","orcid":"","institution":"Alex Ekwueme Federal University","correspondingAuthor":false,"prefix":"","firstName":"Anyigor","middleName":"O.","lastName":"Ogah","suffix":""},{"id":508937979,"identity":"76f02578-d753-4b69-87c4-510ba6371d73","order_by":5,"name":"Iniobong A. Ante","email":"","orcid":"","institution":"PAMO University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Iniobong","middleName":"A.","lastName":"Ante","suffix":""},{"id":508937980,"identity":"659fca72-5490-4db6-8986-d3e41ee1d51d","order_by":6,"name":"Albert E. Okorocha","email":"","orcid":"","institution":"Alex Ekwueme Federal University","correspondingAuthor":false,"prefix":"","firstName":"Albert","middleName":"E.","lastName":"Okorocha","suffix":""}],"badges":[],"createdAt":"2025-08-09 07:53:09","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7332209/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7332209/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":90595527,"identity":"14548c2d-41fc-4157-abd9-2724a3eca80f","added_by":"auto","created_at":"2025-09-04 13:36:20","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":395227,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7332209/v1/29fe350040a5c479d8e950fe.png"},{"id":93041548,"identity":"28cfa98d-8c2f-4526-ad00-528723cd9682","added_by":"auto","created_at":"2025-10-08 12:20:39","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1216708,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7332209/v1/4ed1886b-da34-4485-8882-ae2c45401425.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Molecular docking and in vivo studies of Aloe vera gel for their potential anti-apoptotic activity against aluminium chloride-induced neurotoxicity in mice","fulltext":[{"header":"Introduction","content":"\u003cp\u003eNeurotoxicity, a term that refers to the condition in which the nervous system is directly or indirectly exposed to chemicals either naturally occurring or man-made, thereby impairing the nervous system\u0026rsquo;s normal functioning in both man and animals [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e], [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. As reported by [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e], [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e], the brain is an important organ that is more susceptible to exogenous stimuli than other organs of the body because of numerous polyunsaturated fatty acid contents which makes the neural tissues very sensitive to peroxidation. Several chemicals can elicit neurotoxic disruptions in man, and also many are employed in experimental models for neurotoxicity in animals, such as aluminium chloride.\u003c/p\u003e\u003cp\u003eAluminium chloride is an abundant metal on earth both in the environment and in food, with easy access to the human body via different means [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. It is a well-established neurotoxicant involved in the etiology of neurodegenerative diseases [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. For instance, it has been reported by several researchers that it is a major neurotoxin, disrupter of neurological functions and accelerator of oxidative stress, plaque formation and amyloid beta (Aβ) deposition in animal models, humans and cell lines [\u003cspan additionalcitationids=\"CR9 CR10\" citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]\u0026ndash;[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. According to [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e], apoptosis is the final step in the pathway of toxicity induced by aluminium leading to neurodegeneration. Apoptosis is a controlled form of physiological cell death involving cascades of degenerative events [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Accordingly, experimental and clinical evidence have established that neurological insults, including chemical exposures, can trigger aberrant apoptosis in the brain in vitro or in vivo [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e], [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e], [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. For instance, various apoptotic markers (Bcl-2 and caspase families) have been reported to be expressed following aluminium exposure indicating programmed cell death [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. The specific mechanisms of apoptosis induction vary between exposure paradigms. However, apoptosis is triggered generally by increasing pro-apoptotic and/or decreasing anti-apoptotic markers.\u003c/p\u003e\u003cp\u003eAlternative medicine usage has been in practiced for years as still being used today, even in a more advanced way, including Aloe vera. According to [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e], alternative medicine remains a constituent part of many cultural and technological development around the world. Aloe vera also referred to as \u003cem\u003eAloe Barbadensis Miller\u003c/em\u003e, belonging to the \u003cem\u003eLiliaceae\u003c/em\u003e family is a plant with long-standing reputation majorly for its health, skincare and other benefits [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Research has shown that Aloe vera contains several phytochemicals and about 200 bioactive components, with the inner gel reported to possess most of the components [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. The plant has been reported to possess various pharmacological properties, including antioxidant, antidiabetic, wound healing, anticancer, immune system regulations and memory-enhancing properties [\u003cspan additionalcitationids=\"CR19 CR20\" citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]\u0026ndash;[\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. These potentials were attributed to its bioactive components such as phenolic, anthraquinone, flavonoids, terpenoids and saponins [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Thus, emphasizing its potential as a valuable natural therapeutic agent. Despite all the numerous biological potentials attributed to Aloe vera, no docking-in vivo work has reported on the possible role of Aloe vera in ameliorating apoptosis induced by aluminium chloride in the brain. Following the recent enhancements in search algorithms and energy function, computational designing of a therapeutic agent and molecular docking strategies has become valuable tools in exploring the interaction between proteins and ligands, coupled with inexpensive and streamlined techniques aiding potential therapeutic targets discoveries in specific pathophysiological situations [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. This study therefore, aims to investigate the impact of Aloe vera gel on brain apoptotic and proapoptotic markers of aluminium induced neurotoxicity-mice and explore the biological mechanisms through molecular docking studies.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cp\u003e\u003cstrong\u003e\u003cem\u003eIn vivo study\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eChemical Material\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAluminium Chloride (AlCl₃) was obtained from Sigma-Aldrich (Sigma-Aldrich Cooperation, USA). Every other reagent utilized was of high quality and analytical grade. The reporting of this study as well conforms to ARRIVE 2.0 guidelines [24].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAuthors declare that the study protocol was approved by the Ethics and Animal Handling Committee of the Faculty of Basic Medical Sciences, Alex Ekwueme Federal University, Ndufu-Alike, Nigeria (approval number: AE-FUNAI/FBMS/EAHC/25/012).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePlant Material and extraction\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eLeaves of Aloe vera plant were obtained from Ikwo in Ebonyi State. The leaves were identified and authenticated by Mr. C.B. Udechukwu, a botanist in the Department of Biological Sciences, assigned a voucher specimen number, \u003cem\u003eAE-FUNAI/H/1220 and\u0026nbsp;\u003c/em\u003ewas deposited in the AE-FUNAI Herbarium. Extraction was done by selecting the matured Aloe vera leaves from the base of the plant. The leaves were cleaned and washed of dusts. Then, the thick green skin on both sides was removed thereby exposing the clear gel. The fresh gel was collected by scraping and squeezing into a container. The extraction was freshly done each time of the administration.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eExperimental Animals\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTwenty male Swiss mice (weighing 20-30 grams) were procured from University of Uyo, Akwa Ibom State. The mice were acclimatized for three weeks prior to the study and housed in plastic cages. The animals were exposed to 12-hour light and 12-hour dark cycles. The study was conducted with the animals cared for and humanely treated in accordance with the Guide for the Care and Use of Laboratory Animals.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eExperimental Design\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe mice were randomly allocated to four groups of five rats each (n=5) as follows:\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eControl Group:\u003c/strong\u003e Mice received no treatment (0.5 mL normal saline)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAluminium Chloride-Induced Toxicity Group:\u003c/strong\u003e Mice received aluminium chloride (100 mg/kg body weight, orally) throughout the experimental period\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAloe vera Group:\u003c/strong\u003e Mice were treated with Aloe vera gel (200 mg/kg body weight) daily throughout the experimental period via oral gavage\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAluminium Chloride + Aloe vera Group:\u003c/strong\u003e Mice were exposed to aluminium chloride (100mg/kg) and treated with Aloe vera (200 mg/kg). The treatment protocols lasted for two weeks.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eSample collection\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOn the last day of experimental period, the mice were humanely sacrificed and euthanized by employing ketamine/xylazine (0.1 mL/100 g.bw, intraperitoneally). \u0026nbsp;Following euthanasia, their brains were removed and put in 0.25M cold sucrose buffer solution and were later homogenized for biochemical assays.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eB\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eiochemical assessment\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eEstimation of caspase-3, Bcl-2 and Bax were done quantitatively by standard methods following the manufacturer\u0026rsquo;s guidelines using Caspase Assay Kit; G-Bioscience), (BCL\u0026nbsp;Assay Kit; G-Bioscience) and Bax assay Kit respectively. Results were expressed as \u0026mu;mol/mg protein.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eIn silico study\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003ePreparation of Target Protein and Ligands\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn the present study, the forcefield employed for ligand preparation, protein preparation, and molecular docking was OPLS4. The protein databank (PDB) (http://www.pdb.org) was used to retrieve the crystal structures of proteins, viz., Bax (PDB ID: 5W62), Bcl-2 (PDB ID: P10417) and Casp-3 (PDB ID: P70677). Preparation of the proteins was carried out at a physiological pH range of 7.0 \u0026plusmn; 0.4. The structure of the ligands of Aloe vera which included the Arachidonic acid (PubChem CID: 444899), Beta-carotene (PubChem CID: 5280489), 3-Epi-beta-sitosterol (PubChem CID: 12303645), Aloe-emodin PubChem CID: 10207), Campesterol (PubChem CID: 173183), Aloe resin C (PubChem CID: 11972360), Cholesterol (PubChem CID: 5997), Quercetin PubChem CID: 5280343), and Aloin (PubChem CID: 14989), were retrieved from the PubChem database from NCBI (www.pubchem.ncbi.nlm.nih.gov/). Preparation and protonation of ligands were done at a physiological pH of 7.2. Also, the retrieved structures of the ligands were minimized for their potential interactions with the target protein.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eMolecular docking\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMolecular docking analysis employed extra precision docking (XP) which was used for the retrieved ligands with their corresponding target proteins (BAX, BCL2, CASP3) retrieved from the protein data bank (PDB). The 3D crystal structure of the mouse ligands was complexed with an inhibitor molecule which was eliminated and docking with the ligands was performed. The proteins were energetically minimized and active sites were predicted with the selection of maximized grid parameters. This id followed by calculation of binding energies for the docked complexes. Higher negative binding energy indicates higher binding affinity.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eStatistical Analysis\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe statistical analysis was performed with the help of GraphPad Prism software (GraphPad Software, version 8.0 Inc., San Diego, USA). The results were expressed as Mean \u0026plusmn; Standard Error of Mean (SEM). Analysis of data was done using a one-way analysis of variance (ANOVA) and t-test, utilizing Tukey\u0026rsquo;s post hoc test for comparison of means at 95% (p\u0026lt;0.05) confidence level.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eThe results of all the experiments are presented in Tables 1-3\u0026nbsp;and Figure 1 (a-l) below.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffect of Aloe vera on brain apoptotic and proapoptotic markers in mice\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAs shown in table 1 below, there was a significant increase in Caspase-3 levels in the AlCl₃ group compared to both the Control group (p \u0026lt; 0.05) and the Aloe vera group (p \u0026lt; 0.01). However, treatment with Aloe vera significantly decreased it (p \u0026lt; 0.05). Also, Bax levels were significantly increased in the AlCl₃ group compared to the Aloe vera group and control (p \u0026lt; 0.01). However, there was a significant decline in the Bax levels following treatment with Aloe vera. Meanwhile, Bcl-2 levels significantly decreased in the AlCl₃ group when compared to the Control and Aloe vera groups (p \u0026lt; 0.05), but was significantly alone (p \u0026lt; 0.05) elevated after Aloe vera treatment. In the docking study, Aloe ligands (Aloe resin C, Aloin, Aloe emodin, arachidonic acid and Quercetin) showed favorable binding affinities against Bax, Bcl-2 and Casp-3 as shown in table 3 and figure 1 (a-l) below.\u003c/p\u003e\n\u003cp\u003eTable 1. Effect of Aluminium Chloride and Aloe vera on Brain Apoptotic and Proapoptotic Markers in Mice\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"655\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eParameters\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eControl\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eAlCl\u003csub\u003e3\u003c/sub\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eAloe vera\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eAlCl\u003csub\u003e3\u003c/sub\u003e + Aloe vera\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCasp-3 (µmol/mg)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e22.57±1.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e38.70±0.93\u003cstrong\u003e\u003csup\u003eac*\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e17.60±1.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e33.23±1.48\u003cstrong\u003e\u003csup\u003eabc\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eBax (µmol/mg)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e172.70±19.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e364.30±10.27\u003cstrong\u003e\u003csup\u003ea*c*\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e130.00±3.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e263.00±17.10\u003cstrong\u003e\u003csup\u003eabc\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eBcl-2\u0026nbsp;(µmol/mg)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e7.17±0.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e3.33±0.55\u003cstrong\u003e\u003csup\u003eac\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e12.10±2.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5.30±0.42\u003cstrong\u003e\u003csup\u003ebc\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e(a p\u0026lt;0.05vs control, b p\u0026lt;0.05 vs AlCl\u003csub\u003e3\u003c/sub\u003e, c p\u0026lt;0.05 vs Aloe vera , c* p\u0026lt;0.01 vs Aloe vera; n=5, AlCl\u003csub\u003e3\u003c/sub\u003e= aluminium chloride, Casp-3 = Caspase-3, Bax =Bcl-2 associated X , Bcl-2 =B-cell lymphoma 2\u0026nbsp;)\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2.\u0026nbsp;\u003c/strong\u003ePhytochemicals of \u003cem\u003eAloe vera\u0026nbsp;\u003c/em\u003eligand molecules\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"667\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eMolecule_ID\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eName\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003ePubChem_CID\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eSmiles\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eMOL001439\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eArachidonic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e444899\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eCCCCC/C=C\\C/C=C\\C/C=C\\C/C=C\\CCCC(=O)O\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eMOL002773\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eBeta-carotene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5280489\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eCC1=C(C(CCC1)(C)C)/C=C/C(=C/C=C/C(=C/C=C/C=C(/C=C/C=C(/C=C/C2=C(CCCC2(C)C)C)\\C)\\C)/C)/C\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eMOL000359\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e3-Epi-beta-sitosterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e12303645\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eCC[C@H](CC[C@@H](C)[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2CC=C4[C@@]3(CC[C@H](C4)O)C)C)C(C)C\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eMOL000471\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eAloe-emodin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e10207\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eC1=CC2=C(C(=C1)O)C(=O)C3=C(C2=O)C=C(C=C3O)CO\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eMOL005043\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eCampesterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e173183\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eC[C@H](CC[C@@H](C)C(C)C)[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2CC=C4[C@@]3(CC[C@@H](C4)O)C)C\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eMOL005051\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eAloe resin C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e11972360\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eCC1=CC(=C(C2=C1C(=O)C=C(O2)CC(=O)C)[C@H]3[C@@H]([C@H]([C@@H]([C@H](O3)CO)O)O)OC(=O)/C=C/C4=CC=C(C=C4)O)O[C@H]5[C@@H]([C@H]([C@@H]([C@H](O5)CO)O)O)O\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eMOL000953\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eCholesterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5997\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eC[C@H](CCCC(C)C)[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2CC=C4[C@@]3(CC[C@@H](C4)O)C)C\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eMOL000098\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eQuercetin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5280343\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eC1=CC(=C(C=C1C2=C(C(=O)C3=C(C=C(C=C3O2)O)O)O)O)O\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eMOL005060\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eAloin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e14989\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eC1=CC2=C(C(=C1)O)C(=O)C3=C([C@@H]2[C@H]4[C@@H]([C@H]([C@@H]([C@H](O4)CO)O)O)O)C=C(C=C3O)CO\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eTable 3. Binding energies and interaction details of ligands-proteins complexes.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTarget proteins\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eLigand\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePubChem_ CID\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDocking Score\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eHydrogen Bond\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eGlide energy\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"8\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eBAX (mouse)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eAloe resin C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e11972360\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-8.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-4.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-47.071\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eQuercetin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5280343\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-7.626\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-3.019\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-36.329\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eAloin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e14989\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-4.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-2.128\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-33.171\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eAloe-emodin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e10207\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-3.613\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-1.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-24.227\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCholesterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5997\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-2.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-22.836\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e3-Epi-beta-sitosterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e12303645\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-2.166\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-20.759\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCampesterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e173183\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-2.145\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-20.834\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"9\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eBcl-2 \u0026nbsp;(mouse)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eAloe resin C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e11972360\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-6.908\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-2.534\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-48.76\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eAloe-emodin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e10207\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-6.496\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-1.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-35.466\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eAloin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e14989\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-6.296\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-1.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-40.55\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eQuercetin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5280343\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-6.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-2.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-33.105\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCampesterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e173183\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-5.011\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-0.239\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-19.288\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eArachidonic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e444899\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-4.681\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-29.683\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCholesterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5997\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-4.366\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-20.848\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e3-Epi-beta-sitosterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e12303645\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-3.879\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-24.55\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eBeta-carotene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5280489\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-2.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-37.395\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"10\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eCasp-3 (mouse)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eAloe resin C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e11972360\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-9.811\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-6.074\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-53.796\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eAloin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e14989\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-7.068\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-2.971\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-40.587\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eQuercetin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5280343\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-6.988\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-2.718\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-44.023\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eArachidonic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e444899\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-5.738\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-2.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-35.585\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eAloe-emodin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e10207\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-5.271\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-1.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-30.506\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCholesterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5997\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-4.432\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-0.514\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-26.583\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e3-Epi-beta-sitosterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e12303645\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-3.916\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-1.187\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-24.196\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eCampesterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e173183\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-2.599\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-22.002\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eBeta-carotene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5280489\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-0.693\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e-38.753\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Discussion","content":"\u003cp\u003e\u003cstrong\u003e\u003cem\u003eIn vivo study\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNeurotoxicity exposure has been linked to increased apoptosis in neuronal cells, contributing to various neurodegenerative diseases. For instance, various apoptotic markers have been reported to be expressed following aluminium exposure, indicating programmed cell death. [11] also reported that apoptosis is the final pathway of toxicity induced by aluminium thereby leading to neurodegeneration. Apoptosis plays an important roles in development and tissue homeostasis, and its altered regulation is implicated in degenerative diseases such as Alzheimers’ [25].\u003c/p\u003e\n\u003cp\u003eData from our present study reveals elevated concentration of pro-apoptotic proteins- \u0026nbsp;caspase-3 and Bax in the brain tissues of aluminium chloride–induced AlCl\u003csub\u003e3\u003c/sub\u003e - induced neurotoxic rats with decrease of the anti-apoptotic protein Bcl-2, an indication of enhanced apoptotic signaling and reduced cellular protection against apoptosis following AlCl\u003csub\u003e3\u003c/sub\u003e exposure. Conversely, these results also showed that treatment of neurotoxicant rats with Aloe vera gel reversed the altered Bcl-2 and Bax levels induced by aluminium chloride thereby restoring the balance between regulating proteins of apoptosis.\u003c/p\u003e\n\u003cp\u003eThese findings align with those of previous studies.\u0026nbsp;For instance, a\u0026nbsp;study by [10], reported an aluminium chloride-induced apoptosis within the brain of Wistar rats through the activation of apoptotic caspase-3. Aluminium has been suggested to accelerate deposition of amyloid beta, plaque formation and oxidative stress in rats brain [10]. Similarly, earlier reports by [26] noted that intracisternal infusion of aluminium into the brain of rabbit decreased Bcl-2 and decreased Bax and caspase-3 levels. This also agrees with the reports of [27] in the hippocampal tissues. Increased activation of caspase as reported by [28] confirmed that Aluminium dose-dependently induces apoptosis in brain of rats.\u003c/p\u003e\n\u003cp\u003eTreatment with Aloe vera however, decreased the pro-apoptotic markers and increased the anti-apoptotic as seen in table 1. This also agrees with the study of [4] who reported that Aloe vera gel ameliorated apoptosis together with oxidative stress and hippocampal neuronal loss, in a study aimed at evaluating the protective effects of Aloe vera gel on cisplatin-induced apoptosis in the hippocampus of rats. Thus, resulting in the down-regulation of Casp-3 and Bax, and up-regulation of Bcl-2. Earlier reports indicated that aloe-emodin, one of the active components of Aloe vera could suppress NMDA- induced apoptosis of retinal ganglion cells, thus suggesting its possible role in glaucoma management [29]. Also, \u0026nbsp;recent reports highlighted the capability of aloe sterols to counteracting apoptosis [30], [31]. However, [17] reported the efficacy of Aloe vera in inducing apoptosis in cancer cells and exhibiting immunomodulatory potentials. According to [32], they appear to initiate apoptosis via up-regulation and down-regulation of the expression of P53 and Bcl-2 gene respectively. Another report by [33], also suggested that an extract of Aloe vera exerts anticancer effects on human breast and cervical \u0026nbsp;cancer cells by inhibiting cancer cell growth and inducing apoptosis.\u003c/p\u003e\n\u003cp\u003eThe fate of cell is determined by the measure of balance between pro-apoptotic and anti-apoptotic proteins [34]. When death is favoured by the balance, anti-apoptotic- Bcl-2 inhibits cytochrome c release within the mitochondrial membrane while Bax monitors apoptotic processes by controlling cytochrome c release into the cytoplasm thereby resulting to activation of casp-3 cascade that cleave downstream substrates leading to apoptosis [35], [36]. Chemicals typically cause apoptosis by enhancing pro-apoptotic and/or decreasing anti-apoptotic signaling resulting from oxidative stress, elevated intracellular Ca\u003csup\u003e2+\u003c/sup\u003e levels, or Zn\u003csup\u003e2+\u003c/sup\u003e dyshomeostasis, albeit the precise mechanisms of apoptosis induction differ depending on the exposure paradigm.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eMolecular docking\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAccording to [37], computational screening of natural products have become a necessity due to its easier and cheaper and directional approach. The study involved molecular docking studies of several Aloe vera ligands on proteins involved in apoptosis (Casp-3, Bax and Bcl-2), to assess the binding affinity in terms of binding energy and interaction. In the present study, nine active ligand molecules derived from Aloe vera were performed against the apoptotic proteins. However, five top ligands (aloe emodin, aloin, quercetin, aloe resin C and arachidonic acid; see figure 1) were selected based on their binding affinity and patterns indicated by higher negative docking scores. However, aloe resin C exhibited highest level of interactions with Casp-3, Bax and Bcl-2 active sites. The Bcl-2 family of proteins are key regulators of intracellular apoptotic signal transduction \u0026nbsp;that negate damaging effects of reactive oxygen species and the key biochemical event that defines apoptosis is the activation of caspases [15], [38], [39]. Results of the molecular docking showed that aloe resin C exhibited a H-bonding interactions with Casp3 active sites of GLY 122, ASP 253, SER 205, ARG 207, SER, 63 and SER 120; Bax active sites of LYS 21, ASP 53, GLN 18, PRO 13 and TRP 158; and Bcl2 active site of ALA 146. From molecular docking results, it can be concluded that aloe resin C, aloe emodin, aloin, quercetin, and arachidonic acid interfere with the functions of Casp3, Bax and Bcl2 through H-bonding and hydrophobic interactions, which might suppress the neurodegenerative disease progression. Thereby, portraying the potentials of \u003cem\u003eAloe vera\u003c/em\u003e as a therapeutic agent in enhancing cellular protection and stabilizing neuronal membrane against apoptosis.\u0026nbsp;\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003e\u003cem\u003eIn silico\u003c/em\u003e molecular docking study of active ingredients of \u003cem\u003eAloe vera\u003c/em\u003e with apoptotic proteins together with higher binding energy with all biomarker proteins gave a strong hypothesis to its anti-apopototic potentials. Thus, the findings of this study demonstrated the potentials of \u003cem\u003eAloe vera\u003c/em\u003e as a therapeutic agent in enhancing cellular protection and stabilizing neuronal membrane against apoptotic processes associated with aluminium chloride exposure by enhancing and promoting anti-apoptotic signaling.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding Statement:\u0026nbsp;\u003c/strong\u003eThis research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions:\u0026nbsp;\u003c/strong\u003eStudy conception and design was done by ARN. Material preparation, data collection and analysis were performed by all the authors (ARN, RCN, CSA, EON, AOO, IAA and AEO). The first draft of the manuscript was written and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements:\u0026nbsp;\u003c/strong\u003eThe authors are grateful to Asma’u Bala and Adeshina Odugbemi from the School of Pharmacy, University of Western Cape, South Africa for assisting us with the docking studies.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability statement:\u0026nbsp;\u003c/strong\u003eThe authors declare that the data supporting the findings of the study are available within the article\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclaration of Competing Interest:\u0026nbsp;\u003c/strong\u003eAuthors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBilge, S. Neurotoxicity, Types, Clinical Manifestations, Diagnosis and Treatment. \u003cem\u003eNeurotox. - New. Adv.\u003c/em\u003e 1\u0026ndash;13. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.5772/intechopen.101737\u003c/span\u003e\u003cspan address=\"10.5772/intechopen.101737\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSpencer, P. S. \u0026amp; Lein, P. J. Neurotoxicity, in \u003cem\u003eEncyclopedia of Toxicology: Third Edition\u003c/em\u003e, pp. 489\u0026ndash;500. (2014). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/B978-0-12-386454-3.00169-X\u003c/span\u003e\u003cspan address=\"10.1016/B978-0-12-386454-3.00169-X\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSong, B., Zhou, T., Liu, J. \u0026amp; Shao, L. Q. Involvement of Programmed Cell Death in Neurotoxicity of Metallic Nanoparticles: Recent Advances and Future Perspectives. \u003cem\u003eNanoscale Res. Lett.\u003c/em\u003e \u003cb\u003e11\u003c/b\u003e (484), 1\u0026ndash;12. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/s11671-016-1704-2\u003c/span\u003e\u003cspan address=\"10.1186/s11671-016-1704-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMajd, N. E., Shahraki, R., Tabandeh, M. R. \u0026amp; Hosseinifar, S. Protective effects of Aloe vera gel on cisplatin-induced oxidative stress, apoptosis and neurons structure in rat hippocampus. \u003cem\u003eVet. Res. Forum\u003c/em\u003e. \u003cb\u003e13\u003c/b\u003e (1), 111\u0026ndash;119. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.30466/vrf.2020.119876.2835\u003c/span\u003e\u003cspan address=\"10.30466/vrf.2020.119876.2835\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eClement, L., W, J. T., Ordukenneth, S., D, O. C. \u0026amp; Dirokweni, E. Evaluation of Apoptotic Changes Using BCL-2 on Aluminium Chloride Induced Neurotoxicity Treated with Ethanolic Extract of Carpolobia lutea at Different Doses. \u003cem\u003eAsian J. Med. Princ Clin. Pract.\u003c/em\u003e \u003cb\u003e7\u003c/b\u003e (2), 443\u0026ndash;460 (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eYokel, R. A. \u0026amp; McNamara, P. J. Aluminium Toxicokinetics: An Updated MiniReview. \u003cem\u003ePharmacol. Toxicol.\u003c/em\u003e \u003cb\u003e88\u003c/b\u003e (4), 159\u0026ndash;167. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1600-0773.2001.880401.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1600-0773.2001.880401.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2001).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSkalny, A. V. et al. \u003cem\u003eMolecular mechanisms of aluminum neurotoxicity: Update on adverse effects and therapeutic strategies\u003c/em\u003e 1st edn, Vol. 5 (Elsevier Inc., 2021). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/bs.ant.2020.12.001\u003c/span\u003e\u003cspan address=\"10.1016/bs.ant.2020.12.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBondy, S. C. NeuroToxicology The neurotoxicity of environmental aluminum is still an issue. \u003cem\u003eNeurotoxicology\u003c/em\u003e \u003cb\u003e31\u003c/b\u003e (5), 575\u0026ndash;581. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.neuro.2010.05.009\u003c/span\u003e\u003cspan address=\"10.1016/j.neuro.2010.05.009\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2010).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBlaylock, R. L. Aluminum Induced Immunoexcitotoxicity in Neurodevelopmental and Neurodegenerative Disorders. \u003cem\u003eCurr. Inorg. Chem.\u003c/em\u003e \u003cb\u003e2\u003c/b\u003e (1), 46\u0026ndash;53 (2012).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMesole, S. B., Alfred, O. O., Yusuf, U. A., Lukubi, L. \u0026amp; Ndhlovu, D. Apoptotic Inducement of Neuronal Cells by Aluminium Chloride and the Neuroprotective Effect of Eugenol in Wistar Rats, \u003cem\u003eOxid. Med. Cell. Longev.\u003c/em\u003e, vol. pp. 1\u0026ndash;7, 2020, (2020). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1155/2020/8425643\u003c/span\u003e\u003cspan address=\"10.1155/2020/8425643\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eShahani, L. Aluminium induced Neurotoxicity: A Review. \u003cem\u003ePharmacol. Life Sci. Bull. Env Pharmacol. Life Sci.\u003c/em\u003e \u003cb\u003e9\u003c/b\u003e (3), 63\u0026ndash;71 (2020).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDikranian, K. et al. Apoptosis in the in Vivo Mammalian Forebrain. \u003cem\u003eNeurobiol. Dis.\u003c/em\u003e \u003cb\u003e8\u003c/b\u003e, 359\u0026ndash;379. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1006/nbdi.2001.0411\u003c/span\u003e\u003cspan address=\"10.1006/nbdi.2001.0411\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2001).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMartin, L. J. Neuronal cell death in nervous system development, disease, and injury (Review). \u003cem\u003eInt. J. Mol. Med.\u003c/em\u003e \u003cb\u003e7\u003c/b\u003e, 455\u0026ndash;478 (2001).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSastry, P. S. \u0026amp; Rao, K. S. Apoptosis and the Nervous System. \u003cem\u003eJ. Neurochem\u003c/em\u003e. \u003cb\u003e74\u003c/b\u003e, 1\u0026ndash;20 (2000).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eYuan, J. \u0026amp; Yankner, B. A. Apoptosis in the nervous system, \u003cem\u003eNature\u003c/em\u003e, vol. 407, no. October, pp. 802\u0026ndash;809, (2000).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRanjith, D. Molecular docking studies of aloe vera for their potential antibacterial activity using Argus lab 4.0. 1. \u003cem\u003ePharma Innov. J.\u003c/em\u003e \u003cb\u003e8\u003c/b\u003e (9), 481\u0026ndash;487 (2019).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKaur, S. \u0026amp; Bains, K. Aloe Barbadensis Miller (Aloe Vera) Pharmacological activities and clinical evidence for disease prevention. \u003cem\u003eInt. J. Vitam. Nutr. Res.\u003c/em\u003e \u003cb\u003e94\u003c/b\u003e, 3\u0026ndash;4. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1024/0300-9831/a000797\u003c/span\u003e\u003cspan address=\"10.1024/0300-9831/a000797\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDrudi, D. et al. Aloe barbadensis Miller versus silver sulfadiazine creams for wound healing by secondary intention in dogs and cats: a randomized controlled study. \u003cem\u003eRes. Vet. Sci.\u003c/em\u003e 1\u0026ndash;40. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.rvsc.2017.10.010\u003c/span\u003e\u003cspan address=\"10.1016/j.rvsc.2017.10.010\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSalah, F. et al. Effect of the deacetylation degree on the antibacterial and antibiofilm activity of acemannan from Aloe vera. \u003cem\u003eInd. Crop Prod.\u003c/em\u003e \u003cb\u003e103\u003c/b\u003e, 13\u0026ndash;18. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.indcrop.2017.03.031\u003c/span\u003e\u003cspan address=\"10.1016/j.indcrop.2017.03.031\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFox, L. T. et al. In vitro wound healing and cytotoxic activity of the gel and whole-leaf materials from selected aloe species. \u003cem\u003eJ. Ethnopharmacol.\u003c/em\u003e 1\u0026ndash;27. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.jep.2017.02.017\u003c/span\u003e\u003cspan address=\"10.1016/j.jep.2017.02.017\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRedza-dutordoir, M. \u0026amp; Averill-bates, D. A. Activation of apoptosis signalling pathways by reactive oxygen species. \u003cem\u003eBBA - Mol. Cell. Res.\u003c/em\u003e \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.bbamcr.2016.09.012\u003c/span\u003e\u003cspan address=\"10.1016/j.bbamcr.2016.09.012\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLuty, B. A., Wasserman, Z. R., Stouten, P. F. W. \u0026amp; Hodge, C. N. A Molecular Mechanics / Grid Method for Evaluation of Ligand-Receptor Interactions. \u003cem\u003eJ. Comput. Chem.\u003c/em\u003e \u003cb\u003e16\u003c/b\u003e (4), 454\u0026ndash;464 (1995).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChavda, V., Patel, V. \u0026amp; Patel, S. Investigation of neuroprotective potential of anti-diabetic agents in ischemic brain proteomics through in-silico molecular simulation studies. \u003cem\u003eBiointerface Res. Appl. Chem.\u003c/em\u003e \u003cb\u003e12\u003c/b\u003e (4), 5347\u0026ndash;5362. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.33263/BRIAC124.53475362\u003c/span\u003e\u003cspan address=\"10.33263/BRIAC124.53475362\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePercie Du, N. et al. The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. \u003cem\u003eBMJ Open. Sci.\u003c/em\u003e \u003cb\u003e4\u003c/b\u003e (1), 1\u0026ndash;7. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1136/bmjos-2020-100115\u003c/span\u003e\u003cspan address=\"10.1136/bmjos-2020-100115\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eStrasser, A., Connor, L. O. \u0026amp; Dixit, V. M. Poptosis ignaling. \u003cem\u003eAnnu. Rev. Biochem.\u003c/em\u003e \u003cb\u003e69\u003c/b\u003e, 219\u0026ndash;245 (2000).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGhribi, O., Herman, M. M., Spaulding, N. K. \u0026amp; Savory, J. Lithium inhibits aluminum-induced apoptosis in rabbit hippocampus, by preventing cytochrome c translocation, Bcl-2 decrease, Bax elevation and caspase-3 activation. \u003cem\u003eJ. Neurochem\u003c/em\u003e. \u003cb\u003e82\u003c/b\u003e, 137\u0026ndash;145 (2002).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNiu, Q. \u0026amp; Zhang, H. Relationship between apoptosis of rat hippocampus cells induced by aluminum and the copy of the bcl-2 as well as bax mRNA and the copy of the bcl 2 as well as bax mRNA. \u003cem\u003eJ. Hyg. Res.\u003c/em\u003e \u003cb\u003e34\u003c/b\u003e (6), 671\u0026ndash;675 (2005).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhang, H., Yang, X., Qin, X. \u0026amp; Niu, Q. Caspase-3 is Involved in Aluminum-Induced Impairment of Long- Term Potentiation in Rats Through the Akt / GSK-3 b Pathway. \u003cem\u003eNeurotox. Res.\u003c/em\u003e \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s12640-016-9597-5\u003c/span\u003e\u003cspan address=\"10.1007/s12640-016-9597-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLin, H. et al. Aloe-emodin Suppressed NMDA-induced Apoptosis of Retinal Ganglion Cells through Regulation of ERK Phosphorylation. \u003cem\u003ePhyther Res.\u003c/em\u003e \u003cb\u003e21\u003c/b\u003e, 1007\u0026ndash;1014. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1002/ptr\u003c/span\u003e\u003cspan address=\"10.1002/ptr\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2007).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSaito, M. et al. Oral administration of Aloe vera gel powder prevents UVB-induced decrease in skin elasticity via suppression of overexpression of MMPs in hairless mice. \u003cem\u003eBiosci. Biotechnol. Biochem.\u003c/em\u003e \u003cb\u003e7\u003c/b\u003e, 1416\u0026ndash;1424. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1080/09168451.2016.1156480\u003c/span\u003e\u003cspan address=\"10.1080/09168451.2016.1156480\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMisawa, E. et al. Protective effects of Aloe sterols against UVB-induced photoaging in hairless mice. \u003cem\u003ePhotodermatol Photoimmunol Photomed.\u003c/em\u003e \u003cb\u003e2\u003c/b\u003e, 101\u0026ndash;111. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/ijlh.12426\u003c/span\u003e\u003cspan address=\"10.1111/ijlh.12426\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eShalabi, M. et al. Asian Pacific Journal of Tropical Biomedicine. \u003cem\u003eAsian Pac. J. Trop. Biomed.\u003c/em\u003e \u003cb\u003e5\u003c/b\u003e (5), 375\u0026ndash;381. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/S2221-1691(15)30372-5\u003c/span\u003e\u003cspan address=\"10.1016/S2221-1691(15)30372-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHussain, A., Sharma, C., Khan, S., Shah, K. \u0026amp; Haque, S. Aloe vera Inhibits Proliferation of Human Breast and Cervical Cancer Cells and Acts Synergistically with Cisplatin. \u003cem\u003eAsian Pac. J. Cancer Prev.\u003c/em\u003e \u003cb\u003e16\u003c/b\u003e (7), 2939\u0026ndash;2946 (2015).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBouillet, P. et al. Degenerative Disorders Caused by Bcl-2 Deficiency Prevented by Loss of Its BH3-Only Antagonist Bim. \u003cem\u003eDev. Cell.\u003c/em\u003e \u003cb\u003e1\u003c/b\u003e, 645\u0026ndash;653 (2001).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCai, X., Zhang, H., Tong, D., Tan, Z. \u0026amp; Han, D. Corosolic Acid Triggers Mitochondria and Caspase - dependent Apoptotic Cell Death in Osteosarcoma MG ‐ 63 Cells. \u003cem\u003ePhyther Res.\u003c/em\u003e \u003cb\u003e25\u003c/b\u003e, 1354\u0026ndash;1361 (2011).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMagiera, M. M. et al. Trim17-mediated ubiquitination and degradation of Mcl-1 initiate apoptosis in neurons. \u003cem\u003eCell. Death Differ.\u003c/em\u003e 1\u0026ndash;12. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/cdd.2012.124\u003c/span\u003e\u003cspan address=\"10.1038/cdd.2012.124\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2012).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRifaioglu, A. S. et al. Recent applications of deep learning and machine intelligence on in silico drug discovery: methods, tools and databases, \u003cem\u003eBrief. Bioinform.\u003c/em\u003e, no. August, pp. 1\u0026ndash;35, (2018). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1093/bib/bby061\u003c/span\u003e\u003cspan address=\"10.1093/bib/bby061\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChipuk, J. E., Moldoveanu, T., Llambi, F., Parsons, M. J. \u0026amp; Green, D. R. Review The BCL-2 Family Reunion. \u003cem\u003eMol. Cell.\u003c/em\u003e \u003cb\u003e37\u003c/b\u003e (3), 299\u0026ndash;310. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.molcel.2010.01.025\u003c/span\u003e\u003cspan address=\"10.1016/j.molcel.2010.01.025\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2010).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAkifusa, S. et al. Free Radical Biology \u0026amp; Medicine Globular adiponectin-induced RAW 264 apoptosis is regulated by a reactive oxygen species-dependent pathway involving Bcl-2. \u003cem\u003eFree Radic Biol. Med.\u003c/em\u003e \u003cb\u003e46\u003c/b\u003e (9), 1308\u0026ndash;1316. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.freeradbiomed.2009.02.014\u003c/span\u003e\u003cspan address=\"10.1016/j.freeradbiomed.2009.02.014\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2009).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Docking, Aluminium chloride, Neurotoxicity, Apoptosis, Aloe vera","lastPublishedDoi":"10.21203/rs.3.rs-7332209/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7332209/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eNeurotoxicity resulting from aluminium chloride (AlCl₃) exposure has been linked to increased apoptosis in neuronal cells, contributing to various neurodegenerative diseases. On the other hand, the use of alternative medicines has been practiced for years and formed part of our tradition around the world. This study investigated the impact of Aloe vera gel on brain apoptotic and pro-apoptotic markers in mice subjected to aluminium chloride-induced neurotoxicity via experimental and computational models. Twenty mice were divided into four groups as control, AlCl₃, \u003cem\u003eAloe vera\u003c/em\u003e and AlCl₃ + Aloe vera respectively. Following a two-week treatment period, brain tissues were harvested for quantitative determination of apoptotic markers, employing standard methods. Molecular docking was done on all identified ligands and proteins using a computational approach. The results demonstrated a significant increase in Caspase-3 and Bax levels in the AlCl₃ group compared to both the control and Aloe vera groups. Conversely, Bcl-2 levels were significantly reduced in the AlCl₃ group. Notably, treatment with Aloe vera gel led to decreased Casp-3 and Bax levels in the AlCl₃ + Aloe vera group compared to the AlCl₃ group, while Bcl-2 levels increased. The molecular docking analysis of the present study indicated that Aloe resin C had the best binding score for all the proteins sampled with scores of -8.180, -6.908 and \u0026minus;\u0026thinsp;9.811 for Bax, Bcl-2 and Casp-3 respectively. These findings thus, highlight the potentials of \u003cem\u003eAloe vera\u003c/em\u003e as a therapeutic agent in stabilizing neuronal membrane and mitigating apoptotic processes associated with aluminium chloride exposure by promoting anti-apoptotic signaling.\u003c/p\u003e","manuscriptTitle":"Molecular docking and in vivo studies of Aloe vera gel for their potential anti-apoptotic activity against aluminium chloride-induced neurotoxicity in mice","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-04 13:36:15","doi":"10.21203/rs.3.rs-7332209/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"364066c3-e3d9-42dc-b3c1-3b103ca10fa2","owner":[],"postedDate":"September 4th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":54049002,"name":"Biological sciences/Biochemistry"},{"id":54049003,"name":"Biological sciences/Chemical biology"},{"id":54049004,"name":"Biological sciences/Drug discovery"},{"id":54049005,"name":"Biological sciences/Neuroscience"}],"tags":[],"updatedAt":"2025-10-08T12:12:26+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-04 13:36:15","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7332209","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7332209","identity":"rs-7332209","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}