JQ-1 ameliorates schistosomiasis liver granuloma in mice by suppressing male and female reproductive systems and egg development of Schistosoma japonicum

JQ-1 ameliorates schistosomiasis liver granuloma in mice by suppressing male and female reproductive systems and egg development of Schistosoma japonicum | bioRxiv /* */ /* */ <!-- <!-- /*! * yepnope1.5.4 * (c) WTFPL, GPLv2 */ (function(a,b,c){function d(a){return"[object Function]"==o.call(a)}function e(a){return"string"==typeof a}function f(){}function g(a){return!a||"loaded"==a||"complete"==a||"uninitialized"==a}function h(){var a=p.shift();q=1,a?a.t?m(function(){("c"==a.t?B.injectCss:B.injectJs)(a.s,0,a.a,a.x,a.e,1)},0):(a(),h()):q=0}function i(a,c,d,e,f,i,j){function k(b){if(!o&&g(l.readyState)&&(u.r=o=1,!q&&h(),l.onload=l.onreadystatechange=null,b)){"img"!=a&&m(function(){t.removeChild(l)},50);for(var d in y[c])y[c].hasOwnProperty(d)&&y[c][d].onload()}}var j=j||B.errorTimeout,l=b.createElement(a),o=0,r=0,u={t:d,s:c,e:f,a:i,x:j};1===y[c]&&(r=1,y[c]=[]),"object"==a?l.data=c:(l.src=c,l.type=a),l.width=l.height="0",l.onerror=l.onload=l.onreadystatechange=function(){k.call(this,r)},p.splice(e,0,u),"img"!=a&&(r||2===y[c]?(t.insertBefore(l,s?null:n),m(k,j)):y[c].push(l))}function j(a,b,c,d,f){return q=0,b=b||"j",e(a)?i("c"==b?v:u,a,b,this.i++,c,d,f):(p.splice(this.i++,0,a),1==p.length&&h()),this}function k(){var a=B;return a.loader={load:j,i:0},a}var l=b.documentElement,m=a.setTimeout,n=b.getElementsByTagName("script")[0],o={}.toString,p=[],q=0,r="MozAppearance"in l.style,s=r&&!!b.createRange().compareNode,t=s?l:n.parentNode,l=a.opera&&"[object Opera]"==o.call(a.opera),l=!!b.attachEvent&&!l,u=r?"object":l?"script":"img",v=l?"script":u,w=Array.isArray||function(a){return"[object Array]"==o.call(a)},x=[],y={},z={timeout:function(a,b){return b.length&&(a.timeout=b[0]),a}},A,B;B=function(a){function b(a){var a=a.split("!"),b=x.length,c=a.pop(),d=a.length,c={url:c,origUrl:c,prefixes:a},e,f,g;for(f=0;f<d;f++)g=a[f].split("="),(e=z[g.shift()])&&(c=e(c,g));for(f=0;f<b;f++)c=x[f](c);return c}function g(a,e,f,g,h){var i=b(a),j=i.autoCallback;i.url.split(".").pop().split("?").shift(),i.bypass||(e&&(e=d(e)?e:e[a]||e[g]||e[a.split("/").pop().split("?")[0]]),i.instead?i.instead(a,e,f,g,h):(y[i.url]?i.noexec=!0:y[i.url]=1,f.load(i.url,i.forceCSS||!i.forceJS&&"css"==i.url.split(".").pop().split("?").shift()?"c":c,i.noexec,i.attrs,i.timeout),(d(e)||d(j))&&f.load(function(){k(),e&&e(i.origUrl,h,g),j&&j(i.origUrl,h,g),y[i.url]=2})))}function h(a,b){function c(a,c){if(a){if(e(a))c||(j=function(){var a=[].slice.call(arguments);k.apply(this,a),l()}),g(a,j,b,0,h);else if(Object(a)===a)for(n in m=function(){var b=0,c;for(c in a)a.hasOwnProperty(c)&&b++;return b}(),a)a.hasOwnProperty(n)&&(!c&&!--m&&(d(j)?j=function(){var a=[].slice.call(arguments);k.apply(this,a),l()}:j[n]=function(a){return function(){var b=[].slice.call(arguments);a&&a.apply(this,b),l()}}(k[n])),g(a[n],j,b,n,h))}else!c&&l()}var h=!!a.test,i=a.load||a.both,j=a.callback||f,k=j,l=a.complete||f,m,n;c(h?a.yep:a.nope,!!i),i&&c(i)}var i,j,l=this.yepnope.loader;if(e(a))g(a,0,l,0);else if(w(a))for(i=0;i (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];var j=d.createElement(s);var dl=l!='dataLayer'?'&l='+l:'';j.src='//www.googletagmanager.com/gtm.js?id='+i+dl;j.type='text/javascript';j.async=true;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-M677548'); Skip to main content Home About Submit ALERTS / RSS Search for this keyword Advanced Search New Results JQ-1 ameliorates schistosomiasis liver granuloma in mice by suppressing male and female reproductive systems and egg development of Schistosoma japonicum Jiaming Tian , Bingxin Dai , Li Gong , Pingping Wang , Han Ding , Siwei Xia , Weice Sun , Cuiping Ren , Jijia Shen , View ORCID Profile Miao Liu doi: https://doi.org/10.1101/2022.01.27.477981 Jiaming Tian 1 Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, School of Basic Medical Sciences, Anhui Medical University , 81 # Meishan Road, Hefei, Anhui 230032, People’s Republic of China Find this author on Google Scholar Find this author on PubMed Search for this author on this site Bingxin Dai 1 Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, School of Basic Medical Sciences, Anhui Medical University , 81 # Meishan Road, Hefei, Anhui 230032, People’s Republic of China Find this author on Google Scholar Find this author on PubMed Search for this author on this site Li Gong 1 Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, School of Basic Medical Sciences, Anhui Medical University , 81 # Meishan Road, Hefei, Anhui 230032, People’s Republic of China Find this author on Google Scholar Find this author on PubMed Search for this author on this site Pingping Wang 1 Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, School of Basic Medical Sciences, Anhui Medical University , 81 # Meishan Road, Hefei, Anhui 230032, People’s Republic of China Find this author on Google Scholar Find this author on PubMed Search for this author on this site Han Ding 1 Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, School of Basic Medical Sciences, Anhui Medical University , 81 # Meishan Road, Hefei, Anhui 230032, People’s Republic of China Find this author on Google Scholar Find this author on PubMed Search for this author on this site Siwei Xia 2 The Second Clinical Medical College, Anhui Medical University , 81#Meishan Road, Hefei, Anhui 230032, People’s Republic of China Find this author on Google Scholar Find this author on PubMed Search for this author on this site Weice Sun 2 The Second Clinical Medical College, Anhui Medical University , 81#Meishan Road, Hefei, Anhui 230032, People’s Republic of China Find this author on Google Scholar Find this author on PubMed Search for this author on this site Cuiping Ren 1 Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, School of Basic Medical Sciences, Anhui Medical University , 81 # Meishan Road, Hefei, Anhui 230032, People’s Republic of China Find this author on Google Scholar Find this author on PubMed Search for this author on this site Jijia Shen 1 Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, School of Basic Medical Sciences, Anhui Medical University , 81 # Meishan Road, Hefei, Anhui 230032, People’s Republic of China Find this author on Google Scholar Find this author on PubMed Search for this author on this site For correspondence: iammiaoliu{at}126.com shenjijia{at}hotmail.com Miao Liu 1 Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, School of Basic Medical Sciences, Anhui Medical University , 81 # Meishan Road, Hefei, Anhui 230032, People’s Republic of China Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for Miao Liu For correspondence: iammiaoliu{at}126.com shenjijia{at}hotmail.com Abstract Full Text Info/History Metrics Preview PDF Abstract Schistosomiasis is a serious and widespread parasitic disease caused by infection with Schistosoma japonicum . Because the parasite’s eggs are primarily responsible for schistosomiasis dissemination and pathogenesis, inhibiting egg production is a potential approach to control the spread and severity of the disease. The bromodomain and extra-terminal (BET) proteins represent promising targets for the development of epigenetic drugs against Schistosoma. JQ-1 is a selective inhibitor of the BET protein family. In the present study, JQ-1 was applied S. japonicum in vitro. By using laser confocal scanning microscopy and EdU incorporation assays, we showed that application of JQ-1 to worms in vitro affected egg laying and the development of both the male and female reproductive systems. JQ-1 also inhibited the expression of the reproductive-related genes SjPlk1 and SjNanos1 in S. japonicum . Mice infected with S. japonicum were treated with JQ-1 during egg granuloma formation. JQ-1 treatment significantly reduced the size of the liver granulomas and levels of serum alanine aminotransferase and aspartate aminotransferase in mice and suppressed both egg laying and the development of male and female S. japonicum reproductive systems in vivo. Moreover, the mRNA expression levels of some proinflammatory cytokines were decreased in the parasites. Our findings suggest that JQ-1 treatment attenuates S. japonicum egg– induced hepatic granuloma due at least in part to suppressing the development of the reproductive system and egg production of S. japonicum . These findings further suggest that JQ-1 or other BET inhibitors warrant additional study as a new approach for the treatment or prevention of schistosomiasis. Author summary Among neglected tropical diseases, schistosomiasis is a serious disease caused by infection with the parasite Schistosomiasis japonicum . Treatment of schistosomiasis is currently almost exclusively with praziquantel, which kills mainly adult parasites, with minimal effectiveness against immature schistosomes and eggs. However, the parasite’s eggs are primarily responsible for schistosomiasis dissemination and pathology. In addition, overuse of praziquantel in epidemic areas has led to drug resistance and a reduced cure rate. Thus, new parasite targets for the development of novel therapeutics are crucial. Here, we evaluated the potential of JQ-1, a bromodomain and extra-terminal protein inhibitor, to suppress the production of S. japonicum eggs. Application of JQ-1 to S. japonicum in vitro decreased the number of mature germ cells, the rates of oviposition, and the number of eggs produced in each male-female pairing. JQ-1 treatment of mice infected with S. japonicum ameliorated hepatic granuloma and decreased serum liver enzymes, suggesting improved liver function. These results indicate that JQ-1 inhibits reproductive development and egg production in S. japonicum , providing supporting evidence that JQ-1 warrants additional study for use as a novel approach in the prevention or treatment of schistosomiasis. Introduction Schistosomiasis is an acute and chronic parasitic disease caused by infection with Schistosoma, a parasite that is endemic in 78 countries and is responsible for approximately 280,000 deaths each year [ 1 ]. In China, zoonotic schistosomiasis caused by S. japonicum is major public health threat affecting more than a million people and hundreds of thousands of livestock in China [ 2 ]. Praziquantel is a widely used, high-efficiency, broad-spectrum, oral antiparasitic drug for the treatment of various forms of schistosomiasis, but praziquantel kills only adult worms and is minimally effective against immature schistosomes and eggs [ 2 - 3 ] In addition, the repeated and large-scale use of praziquantel in epidemic areas has led to drug resistance and a reduced cure rate [ 4 - 5 ]. Thus, there is an urgent need to identify new targets for the development of novel parasitic therapeutics. Owing to the key role of fertilized eggs in maintaining the life cycle and inducing pathogenesis [ 2 - 3 ], blocking egg production is a potential alternative approach to control the occurrence, development, and spread of schistosomiasis. The bromodomain and extra-terminal (BET) family of proteins specifically recognizes acetylated lysine residue sites and participates in the regulation of epigenetic protein expression, which plays a key role in regulating various biological processes [ 6 ]. JQ-1 is a selective inhibitor of BET family proteins and has been shown to have promising anti-tumor and anti-inflammatory effects [ 7 ]. In a pilot study, we used JQ-1 to treat hepatic granuloma caused by infection with Schistosoma japonicum . Mice infected with S. japonicum cercariae were injected intraperitoneally with JQ-1 (50 mg/kg) during egg granuloma formation. Unexpectedly, JQ-1 significantly reduced the size of the liver granuloma and the egg burden; however, JQ-1 treatment had no effect on worm load. We hypothesized that JQ-1 would be effective in inhibiting egg production in S. japonicum and sought to learn the mechanisms underlying this effect. Thus, the aim of the present study was to confirm that JQ-1 reduces egg production of S. japonicum and to investigate the potential mechanisms undergirding this effect. To that end, we applied JQ-1 to schistosomes in vitro and assessed the effects on their reproductive development and egg production. We also treated C57BL/6 mice infected with S. japonicum with JQ-1 to assess the effects of the drug on hepatic granuloma and liver function. Our findings indicated that JQ-1 inhibited the reproductive development of males and females and egg production in S. japonicum and ameliorated hepatic granuloma and improved liver function in infected mice. These findings lay a foundation for further study to develop JQ-1 or other BET inhibitors as a new approach for the treatment and prevention of schistosomiasis. Materials and Methods Animals and parasites Female Kunming mice (6-8 weeks old) and female C57BL/6 mice (6-8 weeks old) were provided by the Experimental Animal Center of Anhui Province in Hefei, China. The mice were housed under specific pathogen-free conditions at Anhui Medical University. Oncomelania hupensis snails infected with S. japonicum (a Chinese mainland strain) were purchased from the Jiangxi Provincial Institute of Parasitic Diseases in China. All experiments carried out on animals were conducted in accordance with and were approved by the Animal Ethics Committee of Anhui Medical University (approval No. LLSC20170247). Treatment of schistosomes with JQ-1 in vitro Cercariae were shed in a beaker after exposing 30 O. hupensis infected with S. japonicum to sunlight for 4 h (25–28 °C). For mixed infections, cercariae released from several infected O. hupensis were used. Kunming mice were infected percutaneously with 80–90 cercariae and were humanely killed on the 28th day after infection. All paired parasites were harvested by perfusion and washed three times with RPMI-1640 medium. The worms were then cultured in vitro with RPMI-1640 (Gibco, Grand Island, NY, USA) at 37 °C and 5% CO2. The RPMI-1640 medium was supplemented with 10,000 U/mL penicillin, 10 mg/mL streptomycin, 250 μg/mL amphotericin B (Sangon Biotech, Shanghai, China), 15% fetal calf serum (Gibco), and glutamine (Gibco). For each experiment, 15 pairs of S. japonicum were maintained in a 6-well plate (i.e., 15 pairs/well). JQ-1 (Cat. No. HY-13030, MedChem Express; USA), was dissolved in dimethyl sulfoxide (DMSO). In each experimental group, 15 paired parasites were incubated in 3 mL of medium and treated with different concentrations of JQ-1 (0 μM, 5 μM, 10 μM, and 15 μM). All parasites were cultured at 37 °C for 9 d, and culture media was changed every 24 h. During this time, the viability and morphology of parasites, worm pairings, and the number of eggs were observed and recorded. Confocal laser scanning microscopy (CLSM) For morphological analysis, collected worms were fixed in a solution of alcohol (95%), formalin (3%), and glacial acetic acid (2%)) for at least 24 h. Worms were stained in hydrochloric acid–carmine dye (Ourchem, Shanghai, China) for 17 h and then destained in acidic 70% ethanol until the worms turned light pink. The worms were dehydrated in a graded ethanol series (70%, 90%, and 100%), cleared in 50% xylene diluted in ethanol and 100% xylene for 1 min each, mounted onto slides with neutral gum, sealed with cover glass, and laid flat to dry. The morphology of their reproductive organs was observed with a CLSM (ZEISS LSM 880, Germany) using an emission wavelength of 488 nm. Images were captured and stored at 1024 × 1024 pixels. 5-ethynyl-2′-deoxyuridine (EdU)-incorporation assay For EdU labelling and detection of proliferating cells, paired worms treated with JQ-1 and control worms were incubated with 10 mM of EdU in medium for 24 h. BeyoClick™ EdU-594 Cell Proliferation Kits (Beyotime, Shanghai, China) were used to detect EdU incorporation. Couples were separated, fixed, and stained as described above, with minor alterations. The couples were rinsed twice in PBS and stained with Hoechst 33342 (diluted 1:1000 in PBS) in the dark for 10 min at room temperature. The worms were examined by CLSM using a ZEISS LSM 880 confocal microscope at a wavelength of 405 nm (for Hoechst) and 543 nm (for Azide 594). Treatment of schistosomes with JQ-1 in vivo Four weeks after mice were infected with S. japonicum , mice in the experimental group were injected intraperitoneally with JQ-1 (50 mg/kg body weight per day), and mice in the control group were injected intraperitoneally with vehicle, namely, (2-hydroxypropyl)-β-cyclodextrin (HP-β-CD; Cat. No. 778966, Sigma; USA) 10% (wt/vol), once daily for 15 d. Animals were humanely killed 24 h after the last administration. The parasites, serum, and liver from each mouse were collected for subsequent experimental analyses. Quantitative PCR Total RNA from adult S. japonicum worms or the liver of each mouse was isolated using TRIzol® Reagent (Life Technologies, Carlsbad, CA, USA). The total RNA concentration and purity were detected using a NanoDrop 2000 (Thermo Fisher Scientific, USA). Total RNA (500 ng) from the worms was reverse transcribed into cDNA by using a PrimeScript RT Reagent Kit (TaKaRa, Dalian, China) according to the manufacturer’s instructions. A reliable reference gene for transcriptomic analysis of S. japonicum, PSMD4 , was used as a control gene in the assays [ 8 ] and GAPDH was used as a control gene for transcriptomic analysis of the liver. The experiment was carried out using the StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The relative expression level of each gene was analyzed using SDS v.1.4 software (Applied Biosystems). The procedure for quantitative PCR was conducted as described previously [ 9 ], and the primers were designed and synthesized by Sangon Biotech Co. Ltd. The PCR primer sequences are described in the Supplementary Material. Serum liver enzyme quantification For assessment of mouse liver function, a serum aminotransferase test kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) was used to measure the levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), according to the manufacturer’s instructions. The levels of serum ALT and AST are reported in units per liter. Egg count in liver tissue Approximately 0.1 g of liver tissue was taken from each mouse and weighed. Potassium hydroxide (10%; 1 mL) was added to the liver tissue for digestion at 37 °C for 2 h. The number of eggs in each sample was then counted using a light microscope. Histology and immunohistochemistry of the liver sections Fresh liver tissue (1.0 g) was fixed in 1% buffered formalin and embedded in paraffin. The deparaffinized tissue sections were affixed to slides, and sections (thickness, 4 μm) were stained with hematoxylin and eosin and examined for quantitative and qualitative changes. Computer-assisted morphometric software ((Image-Pro Plus; Media Cybernetics) was used to determine the total areas of the tissue and granuloma on each slide so that the area of the granulomas could be reported as a percentage of the total area for each slide. For each specimen, at least three non-continuous slides were measured, and the mean values obtained from eight mice from each group were used for statistical analysis. Statistical analysis Statistical analysis was performed using GraphPad Prism software (version 6.0). All data were obtained from three independent experiments, each using triplicate samples and following the same protocol. The statistical significance of the difference between two data sets was analyzed using Student’s t-test, and one-way analysis of variance (ANOVA) was used for multiple comparisons, followed by Tukey’s post hoc tests when appropriate. Data are presented as means ± SEM and were considered statistically significant for P-values < 0.05. Results Effects of JQ-1 treatment on pairing rate and egg production The number of male-female paired worms was counted on the 10th day of culture to determine the effect of JQ-1 treatment on the pairing rate. We found that the number of paired worms in the cultures treated with JQ-1 was similar to that in the control group treated with vehicle ( Fig 1A ). No significant changes in schistosome activity or in the number of viable worms were detected between the JQ-1–treated group and the control group. However, the number of eggs collected in the medium and counted using light microscopy was decreased in the cultures treated with JQ-1 compared with controls ( Fig 1 C–F ). To further analyze the effects of JQ-1 on egg production in the paired females, we counted egg numbers and found that compared with the DMSO-treated group, the number of eggs ( P < 0.05) in the JQ-1–treated group decreased in a concentration-dependent manner ( Fig 1B ). Download figure Open in new tab Fig 1. Effect of JQ-1 on male-female pairing rate, egg production, and egg morphology in S. japonicum . Effects of different concentrations (5 μM, 10 μM, and 15 μM) of JQ-1 application on male-female pairing stability (A), egg production (B), and egg morphology (C–F) in S. japonicum pairs cultured in vitro for 10 days. Data represent the mean ± SEM of three independent experiments. Scale bars: 200 μm. Asterisks show statistical differences (*** P < 0.001) tested by one-way ANOVA with multiple comparisons (Tukey’s post-hoc test). JQ-1 treatment decreases mitotic activity in somatic and germ cells We investigated whether JQ-1 affects mitosis in S. japonicum by performing EdU-incorporation assays using JQ-1–treated worms to assess cell proliferation. Worm pairs treated with JQ-1 for 10 d exhibited a substantial decrease in the number of EdU-labeled cells in the gonads, parenchyma, and subtegument of both sexes. In the untreated control group, a substantial number of EdU-labeled cells were detected in the vitellarium and ovary of adult females as well as in the testis and parenchyma of adult males ( Fig 2 ), which indicated high mitotic activity in these organs. Adult worms treated with JQ-1 for 9 d showed a slight decrease in the number of EdU-positive cells in the vitellarium of the females and the testis and parenchyma of the males; greater decreases were observed with increasing concentrations of JQ-1. At the highest concentration, JQ-1–treated worm organs and tissues had almost no EdU-labeled cells ( Fig 2D, H ). Download figure Open in new tab Fig 2. Effect of JQ-1 on cell proliferation in male-female pairs of S. japonicum . Red signals indicate active mitotic cells labeled by EdU; blue signal, Hoechst-positive cells. (A–D) Male S. japonicum and (E–H) female S. japonicum . EdU-incorporated cells are detected in the testes and parenchyma of untreated males (A) and in the vitellarium and ovary of untreated females (E). EdU-positive cells are detected after application of JQ-1 at 5 μM (B, F), 10 μM (C, G), and 15 μM (D, H). Scale bars: 200 μm. Effects of JQ-1 treatment on reproductive organ development Consistent with the observed decreased egg production, CLSM analyses of worm pairs treated with JQ-1 revealed morphologic abnormalities in the gonads of both sexes. After treatment for 10 d, the length and width of the ovaries in females treated with JQ-1 were significantly smaller than those of untreated controls ( Fig 3I–K ). In the control group, no morphological anomalies were observed in the testes of the males ( Fig 4A, E ) or the ovaries of the females ( Fig 3A, E ). Furthermore, the vitellaria of control females contained differentiating vitellocytes. The ovaries of the DMSO-treated female schistosomes were composed of small immature oocytes in the anterior part and larger primary oocytes in the posterior part. The results of CLSM ( Fig 4E–H ) showed that the number of spermatozoa in the seminal vesicles of schistosomes in the JQ-1–treated group was reduced and the development of the spermatozoa was impaired. The testes of DMSO-treated male schistosomes were composed of several testicular lobes arranged bead-like, and each testicular lobe contained a large number of spermatocytes and spermatogonia at different stages. In the group treated with JQ-1, the morphology of whole germ cells in both the testis and ovary were markedly changed. Those changes were more obvious with increasing concentrations of JQ-1. In the ovaries, the sizes of the primary oocytes and immature oocytes were reduced, and the cells of the JQ-1– treated groups were not as full as the cells of DMSO-treated groups ( Fig 3A–D ). The size of the testicular lobes in the group treated with the high concentration of JQ-1 was much smaller than that in the DMSO-treated group, and the numbers of spermatogonia and spermatocytes in the male testes were significantly reduced and more loosely arranged ( Fig 4D ). Large pore-like structures were observed in the testes and ovaries of males and females, respectively ( Figs 3 and 4 , arrows). These morphological changes in both females and males were greatest after treatment with the highest concentration (15 μM) of JQ-1. Compared with controls, the group with JQ-1 treatment showed a markedly reduced diameter of the testicular lobes ( Fig 4I ), which was paralleled by a reduction in cell density within the testes as well as by empty seminal vesicles. Download figure Open in new tab Fig 3. Morphological changes of ovaries and yolk glands in female S. japonicum treated with JQ-1. Worms were stained with carmine hydrochloride and analyzed using confocal laser scanning microscopy. (A, E) Control worms; worms treated with JQ-1 at 5 μM (B, F), 10 μM (C, G), and 15 μM (D, H). Arrows indicate large pore-like structures. Abbreviations: ov, ovary; v, vitellarium; (A–H) Scale bars: 20 μm. Comparison of the length, width, and area of the ovary after JQ-1 application at the indicated concentration (I–K) for 10 d. Data represent the mean ± SEM (n ≥ 15 for each group). Asterisks show statistical differences (** P < 0.05, ** P < 0.01, *** P < 0.001) tested by one-way ANOVA with multiple comparisons. Download figure Open in new tab Fig 4. Morphological changes of spermatozoa in testes and seminal vesicles of S. japonicum treated with JQ-1 in vitro. Worms were stained with carmine hydrochloride and analyzed using confocal laser scanning microscopy. (A, E) Control worms; worms treated with JQ-1 at 5 μM (B, F), 10 μM (C, G), and 15 μM (D, H). Arrows indicate large pore-like structures. (A–D) Scale bars: 20 μm; (E-H) Scale bars: 10 μm. Abbreviations: t, testis; SV, sperm vesicle. (I) Comparison of the diameter of the testicular lobes after JQ-1 application at the indicated concentration for 10 d. Data represent the mean ± SEM (n ≥ 15 for each group). Asterisks show statistical differences (** P < 0.01; *** P < 0.001) tested by one-way ANOVA with multiple comparisons. JQ-1 treatment decreases SjNanos1, SjPlk1 mRNA levels To explore the mechanisms undergirding the observed effects of JQ-1 on S. japonicum , we used quantitative PCR to detect the levels of the S. japonicum protein coding genes polo-like kinase 1 ( SjPlk1 ) and SjNanos1 , two genes related to schistosome reproduction, after application of different concentrations of JQ-1 in vitro. Compared with the control group, the expression levels of SjPlk1 mRNA in the JQ-1–treated worms were down-regulated in both males ( Fig 5A ) and females ( Fig 5B ), and this effect was more marked with increasing concentrations of JQ-1. Similarly, the expression levels of SjNanos1 mRNA were also down-regulated in both males and females, and this effect was also more marked with increasing concentrations of JQ-1. Download figure Open in new tab Fig 5. Results of quantitative PCR analyses of S. japonicum cultured with or without JQ-1 for 10 days. Relative transcription level of Nanos1 in male (A) and female S. japonicum (B). Relative transcription level of Plk1 in male (C) and female (D) S. japonicum . Data represent the mean ± SEM of three independent experiments. Asterisks show statistical differences (* P < 0.05; ** P < 0.01) tested by one-way ANOVA with multiple comparisons. JQ-1 ameliorates liver granuloma caused by S. japonicum infection In the fourth week after S. japonicum infection, mice in the experimental group were injected with JQ-1, and mice in the control group were injected with the vehicle HP-β-CD, once daily for 15 d. All mice were humanely killed after 15 d of treatment ( Fig 6A ). As shown in Fig 6B , livers obtained from mice in the HP-β-CD group had large agglomeration, and granuloma inflammation was severe. However, there was marked reduction of liver surface granulomatous nodules in the JQ-1–treated group. The livers obtained from mice in the JQ-1-treated group were lighter and more vivid in color, and the surface was relatively smooth compared with the livers from mice in the control group. Hematoxylin and eosin staining of the liver showed that the percentage of the area of the liver that had granulomas in the JQ-1–treated group was significantly reduced compared with that in the HP-β-CD control group ( Fig 6C ) (P < 0.05). In addition, the weights of the liver and spleen obtained from mice treated with JQ-1 were significantly lower than those from control mice ( Fig 6D ). Moreover, the results of the AST and ALT assays showed that the activity of serum transaminase in the JQ-1– treated group was significantly lower than that in the control group ( Fig 6F ) ( P < 0.05). Download figure Open in new tab Fig 6. Effect of JQ-1 treatment on liver granuloma in mice infected with S. japonicum . (A) Protocol used to assess liver granuloma in mice. (B) Gross appearance of livers obtained from mice infected with S. japonicum and treated with JQ-1 or vehicle (HP-β-CD). Liver slices stained with hematoxylin and eosin. Scale bars: 500 μm. (C) Measurement of granuloma area as a percentage of total area as assessed by computer-aided morphometry. (D) Liver weight of S. japonicum –infected mice treated with JQ-1 or HP-β-CD. (E) Spleen weight of S. japonicum –infected mice treated with JQ-1 or HP-β-CD. (F) Effect of JQ-1 treatment on serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in mice infected with S. japonicum . Data represent the mean ± SEM (n = 9 for each group). Asterisks denote statistically significant differences (Student’s t-test, * P < 0.05; *** P < 0.001) vs. the HP-β-CD–treated control group. ns, not significant. To further explore the effect of JQ-1 treatment to ameliorate hepatic granuloma in mice infected with S. japonicum , we used quantitative PCR to detect the expression levels of a series of inflammatory factors. The mRNA expression levels of the genes in the HP-β-CD–treated control group were set at 1. As shown in Fig 7 , the mRNA expression levels of the inflammatory factors in the JQ-1–treated group relative to those in the control group were significantly decreased ( P < 0.05). Notably, the expression level of interleukin 13 (IL-13), an inflammatory factor closely related to the formation of granuloma caused by S. japonicum , was significantly reduced. Download figure Open in new tab Fig 7. Effect of JQ-1 treatment on mRNA expression of inflammatory-related genes in the liver of mice infected with S. japonicum . The mRNA levels are expressed relative to those in controls following normalization with GAPDH. Data represent the mean ± SEM (n = 9 for each group). Asterisks denote statistically significant differences (Student’s t test, * P < 0.05). Effects of JQ-1 treatment on schistosome eggs in the liver and on adult worms in mice infected with S. japonicum The above results suggested that JQ-1 alleviated liver injury caused by schistosome infection to some extent and reduced the formation of hepatic granuloma in mice. To observe whether JQ-1 affected S. japonicum eggs in the liver, we evaluated the quantity of eggs in the liver of mice in the JQ-1–treated group compared with that in the HP-β-CD–treated control group after schistosome infection. The liver tissue obtained following digestion with 10% potassium hydroxide was used to observe the morphology of the eggs and to count them. We found that the proportion of abnormally small or dead eggs was increased in the JQ-1–treated group ( Fig 8 ). The volume of eggs in the liver of JQ-1–treated infected mice was approximately 40% lower than that of control mice injected with HP-β-CD. By contrast, the numbers of adult worms and worm pairs in the livers of the JQ-1–treated group were not affected. The percentage of the liver that was granuloma tissue in the JQ-1–treated group was significantly decreased compared with control. Although this effect in the treated group may have been due to the significant decrease in the number of eggs or to the increase in the number of small or dead eggs, it may also be related to the immune regulation of JQ-1 in mice. We used an EdU-incorporation assay to assess the proliferation of germ cells in schistosomes of infected mice ( Fig 8 ). Although some differences between the control group and the treated group were observed, the differences were not as obvious as those observed in the in vitro experiments. Download figure Open in new tab Fig 8. JQ-1 treatment alters germ cell proliferation of S. japonicum and egg production in the liver of mice infected with S. japonicum . (A) Egg morphology and (B) production in the liver. (C) Numbers of adult worms and (D) worm pairs in the liver. Red signals indicate active mitotic cells labeled by EdU; blue, Hoechst-positive cells. EdU-incorporated cells in control worms were detected in the testes and parenchyma of males (E) and in the vitellarium and ovary of females (F). (G, H) EdU-positive cells detected in S. japonicum of mice treated with JQ-1. (A) Scale bars: 500 μm. (E-H) Scale bars: 100 μm. Data represent the mean ± SEM (n = 9 for each group). Asterisks denote statistically significant differences (Student’s t test, ** P < 0.01). CLSM analyses of the JQ-1–treated group revealed morphologic abnormalities in the gonads of both sexes. In the control HP-β-CD–treated group, no morphological anomalies were observed in the ovaries of the females ( Fig 9A ) or the testes of the males (Fig 10B). By contrast, compared with the control group, the number of spermatozoa in the seminal vesicles of schistosomes in the JQ-1–treated group was reduced and the development of spermatozoa was impaired ( Fig 9C, F ). In addition, the overall morphology of the germ cells of schistosomes in both the testis and ovary were markedly changed. The sizes of the primary oocytes and immature oocytes were reduced, and the cells in the JQ-1–treated group were not as filled as the cells in the HP-β-CD group ( Fig 9A, D ). Moreover, large pore-like structures could be found in the testes and ovaries of male and female schistosomes, respectively, in the JQ-1–treated group (Fig 10, arrows). Download figure Open in new tab Fig 9. Morphological changes in the testis and ovary of S. japonicum treated with JQ-1 in vivo. Worms were stained with carmine hydrochloride and analyzed using confocal laser scanning microscopy. (A–C) Testes and seminal vesicles of worms in control mice. (D–F) Testes and seminal vesicles of worms in mice treated with JQ-1. Abbreviations: ov, ovary; t, testes; SV, seminal vesicles. Scale bars: 20 μm. Discussion The present study assessed the effects of JQ-1 application on S. japonicum in vitro and in vivo and investigated the potential mechanisms undergirding the observed effects. The results of our in vitro studies indicated that although JQ-1 application did not affect the number or pairing of adult schistosomes, the number of eggs decreased in a concentration-dependent manner. In addition, mitotic activity in the somatic and germ cells of the adult worms decreased. The numbers of spermatogonia and spermatocytes were significantly decreased and the testicular lobes were significantly smaller in male schistosomes treated with JQ-1 compared with schistosomes in the control group. Moreover, large pore-like structures were observed in the testes and ovaries of JQ-1–treated schistosomes. These results suggested that JQ-1 specifically inhibited the proliferation of germ cells. Our EdU incorporation assays confirmed that JQ-1 reduced the number of proliferating cells in both the ovaries and testes of schistosomes. Proliferation of those cells is essential for the initiation and continuous production of mature germ cells. Treatment with JQ-1 also decreased the expression levels of two genes related to schistosome reproduction, SjPlk1 and SjNanos1 , in a concentration-dependent manner. Thus, this study is the first, to our knowledge, to show that JQ-1 is effective against reproductive development and egg production of adult S. japonicum in vitro. In schistosomiasis in humans, morbidity is mainly attributed to the eggs because of the granulomatous inflammatory reaction caused by the host immune response to egg antigens [ 2 - 3 ]. Thus, we assessed the ability of JQ-1 to treat hepatic granuloma in mice infected with S. japonicum in vivo. JQ-1 treatment significantly decreased the percentage of the area of the liver with granulomas, the activity of liver serum transaminase, and schistosome egg production in the liver of mice without affecting the survival of adult worms. The attenuated egg production was accompanied by decreased expression levels of proinflammatory cytokines, which may have contributed to the amelioration of hepatic granuloma. Taken together, our findings provide evidence supporting the development of JQ-1 as an anti-schistosomal agent. The BET family proteins are characterized by the presence of two tandem bromodomains and an extra-terminal domain, which are found in BRD2, BRD3, BRD4, and BRDT in mammalians [ 6 ] The domain organization of mammalian BET proteins is conserved in orthologs, including in Drosophila FSH and Saccharomyces cerevisiae Bdf1 and Bdf2. Bromodomains that specifically bind acetylated lysine residues in histones serve as chromatin-targeting modules that decipher the histone acetylation code. BET proteins play a crucial role in regulating gene transcription through epigenetic interactions between bromodomains and acetylated histones during cell proliferation and differentiation [ 10 - 11 ]. Brd2 mRNA is express in distinct patterns during ovarian folliculogenesis, which is essential for embryonic development in the mouse [ 12 - 13 ], Brdt acetylated histone H4-dependent chromatin remodeling in mammalian spermiogenesis is essential for male germ cell differentiation [ 14 - 15 ]. In addition, a BRDT-like function in Drosophila plays crucial roles in spermatid differentiation [ 16 ]. Epigenetic modifications, including DNA methylation, histone modifications, and non-coding RNAs, play important roles in the development and reproduction of schistosomes [ 17 ]. SmGCN5 and SmCBP1are two histone acetyltransferases of S. mansio n, the knockdown of SmGCN5 or SmCBP1 significantly inhibited Smp14 expression, which compromised the reproductive system of mature females, egg-laying and egg morphology [ 18 ]. Sirtuins are a family of histone deacetylases, and sirtuin inhibitors can inhibit apoptosis and death in schistosome larvae, the disruption of adult worm pairs, inhibition of egg laying and damage to the male and female worm reproductive systems [ 19 - 20 ]. As a first-in-class potent and selective inhibitor of the BET signaling pathway, JQ-1 has been widely used in biology studies. The results of some of those many studies indicate that JQ-1 interacts with the BRD pocket in a manner competitive with acetylated peptide binding, resulting in the displacement of BET proteins from acetylated chromatin in cells exposed to these inhibitors along with their associated transcript initiation and elongation factors. JQ-1 has also been used as a pharmacological tool for elucidating the roles and functions of BET in mammals. However, little is known about the effect of JQ-1 on parasites. Nanos has been described as a necessary factor in the differentiation and migration of primordial germ cells, which play an essential role in the proliferation of germ cells in schistosomes [ 21 - 22 ]. SmPlk1 regulates the cell cycle G2/M transition in Xenopus oocytes, which is important for cell-cycle progression in the gonadal cells of Schistosoma [ 23 - 24 ]. In the present study, we investigated whether JQ-1 also affected the transcript level of Nanos1 and Plk1 . Indeed, treatment with JQ-1 significantly reduced the transcript level of both these genes in male and female worms, which likely affected the proliferation of the gonadal cells in Schistosoma. This study has limitations that should be considered when interpreting our results. On the basis of previous publications [ 25 - 26 ], we used only a single dose of JQ-1 (50 mg/kg) to treat mice infected with S. japonicum for 15 days. Thus, we were unable to make any comparisons of the effects after various treatment times or dosage on parasites in infected mice. Future studies are needed to find the therapeutic optimum dosage. In conclusion, our data showed that JQ-1 treatment ameliorated S. japonicum egg– induced hepatic granuloma, which may be due in part to suppressing the development of both the male and female reproductive systems and female egg production in this parasite. Our findings provide theoretical and practical evidence supporting the development of JQ-1 as an anti-schistosomal agent. Funding Statement This work was supported by grants from the National Natural Science Foundation of China ( http://www.nsfc.gov.cn ) (grant numbers 81271865) and Key University Science Research Project of Anhui Province of China (KJ2019A0223). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Data Availability All relevant data are within the manuscript and its Supporting Information files. Acknowledgments We thank a native English speaker (Science/Medical English Editing for International Researchers) for modifying the manuscript. References 1. ↵ LoVerde PT . Schistosomiasis . Adv Exp Med Biol . 2019 ; 1154 : 45 – 70 . doi: 10.1007/978-3-030-18616-6_3 . PMID: 31297759 . OpenUrl CrossRef PubMed 2. ↵ McManus DP , Dunne DW , Sacko M , Utzinger J , Vennervald BJ , Zhou XN . Schistosomiasis . Nat Rev Dis Primers . 2018 ; 4 ( 1 ): 13 . doi: 10.1038/s41572-018-0013-8 . PMID: 30093684 . OpenUrl CrossRef PubMed 3. ↵ Colley DG , Bustinduy AL , Secor WE , King CH . Human schistosomiasis . Lancet . 2014 ; 383 ( 9936 ): 2253 – 64 . doi: 10.1016/S0140-6736(13)61949-2 . PMID: 24698483 . PMCID: PMC4672382 . OpenUrl CrossRef PubMed Web of Science 4. ↵ Melman SD , Steinauer ML , Cunningham C , Kubatko LS , Mwangi IN , Wynn NB , et al. Reduced susceptibility to praziquantel among naturally occurring Kenyan isolates of Schistosoma mansoni . PLoS Negl Trop Dis . 2009 , 3 ( 8 ): 504 . doi: 10.1371/journal.pntd.0000504 . PMID: 19688043 . PMCID: PMC2721635 . OpenUrl CrossRef PubMed 5. ↵ Kasinathan RS , Greenberg RM . Pharmacology and potential physiological significance of schistosome multidrug resistance transporters . Exp Parasitol , 2012 , 132 ( 1 ): 2 – 6 . doi: 10.1016/j.exppara.2011.03.004 . PMID: 21420955 . PMCID: PMC3154572 . OpenUrl CrossRef PubMed 6. ↵ Taniguchi Y. The Bromodomain and Extra-Terminal Domain (BET) Family: Functional Anatomy of BET Paralogous Proteins . Int J Mol Sci , 2016 , 17 ( 11 ): E1849 . doi: 10.3390/ijms17111849 . PMID: 27827996 . PMCID: PMC5133849 . OpenUrl CrossRef PubMed 7. ↵ Ferri E , Petosa C , McKenna CE . Bromodomains: Structure, function and pharmacology of inhibition . Biochem Pharmacol . 2016 Apr 15; 106 : 1 – 18 . doi: 10.1016/j.bcp.2015.12.005 . Epub 2015 Dec 18. PMID: 26707800 . OpenUrl CrossRef PubMed 8. ↵ Liu S , Cai P , Hou N , Piao X , Wang H , Hung T , et al. Genome-wide identification and characterization of a panel of house-keeping genes in Schistosoma japonicum . Mol Biochem Parasitol . 2012 Mar-Apr ; 182 ( 1-2 ): 75 – 82 . doi: 10.1016/j.molbiopara.2011.12.007 . Epub 2012 Jan 8. PMID: 22245333 . OpenUrl CrossRef PubMed 9. ↵ Liu F , Ding H , Tian J , Zhou C , Yang F , Shao W , et al. Differential gene expression, including Sjfs800, in Schistosoma japonicum females at pre-pairing, initial pairing and oviposition . Parasit Vectors . 2019 Aug 23; 12 ( 1 ): 414 . doi: 10.1186/s13071-019-3672-8 . PMID: 31443730 ; PMCID: PMC6708146 OpenUrl CrossRef PubMed 10. ↵ Doroshow DB , Eder JP , LoRusso PM . BET inhibitors: a novel epigenetic approach . Ann Oncol . 2017 Aug 1; 28 ( 8 ): 1776 – 1787 . doi: 10.1093/annonc/mdx157 . PMID: 28838216 . OpenUrl CrossRef PubMed 11. ↵ Cheung KL , Kim C , Zhou MM . The Functions of BET Proteins in Gene Transcription of Biology and Diseases . Front Mol Biosci . 2021 Sep 3; 8 : 728777 . doi: 10.3389/fmolb.2021.728777 . PMID: 34540900 ; PMCID: PMC8446420 . OpenUrl CrossRef PubMed 12. ↵ Trousdale RK , Wolgemuth DJ . Bromodomain containing 2 (Brd2) is expressed in distinct patterns during ovarian folliculogenesis independent of FSH or GDF9 action . Mol Reprod Dev , 2004 , ( 3 ): 261 – 268 . doi: 10.1002/mrd.20059 . PMID: 15112318 . OpenUrl CrossRef PubMed Web of Science 13. ↵ Shang E , Wang X , Wen D , Greenberg DA , Wolgemuth DJ . Double bromodomain-containing gene Brd2 is essential for embryonic development in mouse . Dev Dyn , 2009 , 238 ( 4 ): 908 – 917 . doi: 10.1002/dvdy.21911 . PMID: 19301389 . PMCID: PMC2771124 . OpenUrl CrossRef PubMed Web of Science 14. ↵ Berkovits BD , Wolgemuth DJ . The first bromodomain of the testis-specific double bromodomain protein Brdt is required for chromocenter organization that is modulated by genetic background . Developmental Biolog , 2011 , 360 ( 2 ): 358 – 368 . doi: 10.1016/j.ydbio.2011.10.005 . PMID: 22020252 . PMCID: PMC3217133 . OpenUrl CrossRef PubMed 15. ↵ Matzuk MM 1, McKeown MR , Filippakopoulos P , Li Q , Ma L , Agno JE , et al. Small-molecule inhibition of BRDT for male contraception . Cell , 2012 , 150 ( 4 ): 673 – 684 . doi: 10.1016/j.cell.2012.06.045 . PMID: 22901802 . PMCID: PMC3420011 . OpenUrl CrossRef PubMed Web of Science 16. ↵ Kimura S , Loppin B. Two bromodomain proteins functionally interact to recapitulate an essential BRDT-like function in Drosophila spermatocytes . Open Biol . 2015 , 5 ( 2 ): 140145 . doi: 10.1098/rsob.140145 . PMID: 25652540 . PMCID: PMC4345279 . OpenUrl CrossRef PubMed 17. ↵ Liu W. Epigenetics in Schistosomes: What We Know and What We Need Know . Front Cell Infect Microbiol . 2016 Nov 11; 6 : 149 . doi: 10.3389/fcimb.2016.00149 . PMID: 27891322 ; PMCID: PMC5104962 OpenUrl CrossRef PubMed 18. ↵ Carneiro VC , de Abreu da Silva IC , Torres EJ , Caby S , Lancelot J , Vanderstraete M , et al. Epigenetic changes modulate schistosome egg formation and are a novel target for reducing transmission of schistosomiasis . PLoS Pathog . 2014 May 8; 10 ( 5 ): e1004116 . doi: 10.1371/journal.ppat.1004116 . PMID: 24809504 ; PMCID: PMC4014452 . OpenUrl CrossRef PubMed 19. ↵ Lancelot J , Caby S , Dubois-Abdesselem F , Vanderstraete M , Trolet J , Oliveira G , et al. Schistosoma mansoni Sirtuins: characterization and potential as chemotherapeutic targets . PLoS Negl Trop Dis . 2013 Sep 12; 7 ( 9 ): e2428 . doi: 10.1371/journal.pntd.0002428 . PMID: 24069483 ; PMCID: PMC3772001 . OpenUrl CrossRef PubMed 20. ↵ Lancelot J , Cabezas-Cruz A , Caby S , Marek M , Schultz J , Romier C , et al. Schistosome sirtuins as drug targets . Future Med Chem . 2015 ; 7 ( 6 ): 765 – 82 . doi: 10.4155/fmc.15.24 . PMID: 25996069 . OpenUrl CrossRef PubMed 21. ↵ Liu Q , Zhu L , Liu F , Hua M , Ding H , He S , et al. Function of Nanos1 gene in the development of reproductive organs of Schistosoma japonicum . Parasitol Res . 2017 May ; 116 ( 5 ): 1505 – 1513 . doi: 10.1007/s00436-017-5427-9 . Epub 2017 Mar 28. PMID: 28352943 . OpenUrl CrossRef PubMed 22. ↵ Kellershohn J , Thomas L , Hahnel SR , Grünweller A , Hartmann RK , Hardt M , et al. Insects in anthelminthics research: Lady beetle-derived harmonine affects survival, reproduction and stem cell proliferation of Schistosoma mansoni . PLoS Negl Trop Dis . 2019 Mar 14; 13 ( 3 ): e0007240 . doi: 10.1371/journal.pntd.0007240 . PMID: 30870428 ; PMCID: PMC6436750 . OpenUrl CrossRef PubMed 23. ↵ Long T , Neitz RJ , Beasley R , Kalyanaraman C , Suzuki BM , Jacobson MP , et al. Structure-Bioactivity Relationship for Benzimidazole Thiophene Inhibitors of Polo-Like Kinase 1 (PLK1), a Potential Drug Target in Schistosoma mansoni . PLoS Negl Trop Dis . 2016 Jan 11; 10 ( 1 ): e0004356 . doi: 10.1371/journal.pntd.0004356 . PMID: 26751972 ; PMCID: PMC4709140 . OpenUrl CrossRef PubMed 24. ↵ Long T , Vanderstraete M , Cailliau K , Morel M , Lescuyer A , Gouignard N , et al. SmSak, the second Polo-like kinase of the helminth parasite Schistosoma mansoni: conserved and unexpected roles in meiosis . PLoS One . 2012 ; 7 ( 6 ): e40045 . doi: 10.1371/journal.pone.0040045 . Epub 2012 Jun 29. PMID: 22768216 ; PMCID: PMC3386946 . OpenUrl CrossRef PubMed 25. ↵ Ding N , Hah N , Yu RT , Sherman MH , Benner C , Leblanc M , et al. BRD4 is a novel therapeutic target for liver fibrosis . Proc Natl Acad Sci U S A . 2015 Dec 22; 112 ( 51 ): 15713 – 8 . doi: 10.1073/pnas.1522163112 . Epub 2015 Dec 7. PMID: 26644586 ; PMCID: PMC4697417 . OpenUrl Abstract / FREE Full Text 26. ↵ Filippakopoulos P , Qi J , Picaud S , Shen Y , Smith WB , Fedorov O , et al. Selective inhibition of BET bromodomains . Nature . 2010 Dec 23; 468 ( 7327 ): 1067 – 73 . doi: 10.1038/nature09504 . Epub 2010 Sep 24. PMID: 20871596 ; PMCID: PMC3010259 . OpenUrl CrossRef PubMed Web of Science Back to top Previous Next Posted January 27, 2022. Download PDF Email Thank you for your interest in spreading the word about bioRxiv. NOTE: Your email address is requested solely to identify you as the sender of this article. Your Email * Your Name * Send To * Enter multiple addresses on separate lines or separate them with commas. 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Share JQ-1 ameliorates schistosomiasis liver granuloma in mice by suppressing male and female reproductive systems and egg development of Schistosoma japonicum Jiaming Tian , Bingxin Dai , Li Gong , Pingping Wang , Han Ding , Siwei Xia , Weice Sun , Cuiping Ren , Jijia Shen , Miao Liu bioRxiv 2022.01.27.477981; doi: https://doi.org/10.1101/2022.01.27.477981 Share This Article: Copy Citation Tools JQ-1 ameliorates schistosomiasis liver granuloma in mice by suppressing male and female reproductive systems and egg development of Schistosoma japonicum Jiaming Tian , Bingxin Dai , Li Gong , Pingping Wang , Han Ding , Siwei Xia , Weice Sun , Cuiping Ren , Jijia Shen , Miao Liu bioRxiv 2022.01.27.477981; doi: https://doi.org/10.1101/2022.01.27.477981 Citation Manager Formats BibTeX Bookends EasyBib EndNote (tagged) EndNote 8 (xml) Medlars Mendeley Papers RefWorks Tagged Ref Manager RIS Zotero Tweet Widget Facebook Like Google Plus One Subject Area Microbiology Subject Areas All Articles Animal Behavior and Cognition (8003) Biochemistry (18709) Bioengineering (14841) Bioinformatics (44364) Biophysics (22561) Cancer Biology (19682) Cell Biology (26845) Clinical Trials (138) Developmental Biology (13943) Ecology (20970) Epidemiology (2067) Evolutionary Biology (25415) Genetics (16151) Genomics (23478) Immunology (18681) Microbiology (42421) Molecular Biology (18022) Neuroscience (93355) Paleontology (699) Pathology (2977) Pharmacology and Toxicology (5086) Physiology (8105) Plant Biology (15969) Scientific Communication and Education (2094) Synthetic Biology (4549) Systems Biology (10219) Zoology (2384) window.__CF$cv$params={r:'a3c71a457e22c9e7',t:'MTc4OTYzNzUzNQ==',u:'01a0aeb5b11577218422af860a5091e7',ut:'pCzx4l4CgyyRd8TF0cgGmuDHA3IUEYp1.lgMqshpR58-1789637538-1.2.1.1-KxoCOm.2OWKMqEaaKfb69y9rilWVgN_SQkRjd6cgSz6zXeomXnhSgQz_tULAdCLsXGedSbpiKhfBOSI36DZnN8mkzOmIcZjPRmGskqWSRYU',i:60};(function(){if(!document.body)return;var s=document.createElement('script');s.src='/cdn-cgi/challenge-platform/scripts/precursor/main.js';document.head.appendChild(s);})();