Pharmacokinetics, tissue distribution, effect on proliferation inhibition and apoptosis of hepatic stellate cells and molecular mechanism of Suc-GTS- lip which is glycyrrhetinic acid derivative receptor-mediated liver- targeting liposome

Objective: To investigate the liver targeting effect of salvianolic acid B (Sal B)-tanshinone IIA (TSN)-glycyrrhetinic acid (GA) liposomes (GTS-lip) with the incorporation of 3-succinic-30-stearyl glycyrrhetinic acid (18-GA-Suc)and To confirm the efficacy of it in treating hepatic fibrosis at the cellular level. Method 18-GA-Suc was inserted into GTS-lip to prepare GA-derived receptor-mediated active targeting liposomes (Suc-GTS-lip). Compared the pharmacokinetics and tissue distribution of Suc-GTS-lip with GTS-lip.The proliferative inhibition rate of hepatic stellate cell (HSC) affected by Suc-GTS-lip was determinded by MTT assay. The apoptosis rate of HSC was measured with Annexin V-FITC / PI method using flow cytometry. The expression levels of MMP-1, TIMP- 1, TIMP-2, Collagen-Ⅰ and Collagen-Ⅲ mRNA in HSC were determined by RT-PCR. Result The results of the pharmacokinetic study showed that incorporation of ligands changed the bioavailability of the three liposomal drugs in mice. Tissue distribution results showed that the AUC and Cmax of Sal B were increased in Suc-GTS-lip compared with GTS-lip. The liver targeting effect of Sal B, which was entrapped in the aqueous phase, was achieved with the help of ligands. The in vivo imaging study showed that the modified liposomes tended to accumulate in the liver, with the liver fluorescence intensity reaching its peak at 30 min. The MTT test results showed that Suc-GTS-lip could significantly inhibit the proliferation of HSC and high, medium, low dose groups of Suc-GTS-lip induced apoptosis rates of HSC, which were 75.23 ± 2.56%, 27.60 ± 0.95%, 20.77 ± 3.97%.The RT-PCR results showed that, compared with the blank control group, the Suc-GTS-lip could increase matrix metalloproteinases (MMP-1), high-dose, medium-dose group(P < 0.001), low-dose group(P < 0.01);decrease tissue inhibitor of matrix metalloproteinases (TIMP-1 and TIMP-2), high-dose group(P < 0.001), medium-dose group(P < 0.01); and lessen synthesis of collagen-Ⅰand collagen-Ⅲ, high-dose, medium-dose group(P < 0.001), low-dose group(P < 0.05), suggesting that Suc-GTS-lip may induce the apoptosis of HSC from many ways. Conclusion Suc-GTS-lip could therefore promote the -stabilities of the drugs and enhance their liver targeting ability.The efficacy of Suc-GTS-lip in the treatment of hepatic fibrosis was confirmed from the cellular level, which provided a basis for the animal efficacy in vivo.