甘草次酸衍生物受体介导的复方脂质体肝靶向性研究及对肝星状细胞的影响

Liver targeting of compound liposomes mediated by glycyrrhetinic acid derivative receptor and its effect on hepatic stellate cells

将3-琥珀酸-30-硬脂基甘草次酸(18-GA-Suc)插入甘草次酸(GA)-丹参酮Ⅱ_(A)(TSN)-丹酚酸B(Sal B)脂质体(GTS-lip),制备甘草次酸(GA)受体介导的肝靶向复方脂质体(Suc-GTS-lip);通过UPLC比较Suc-GTS-lip与GTS-lip的药代动力学和组织分布,并对Suc-GTS-lip进行活体成像追踪;考察Suc-GTS-lip对肝星状细胞(HSC)增殖抑制的影响,并探究其改善肝纤维化的分子机制。药代动力学研究结果表明,Sal B的AUC由(636.06±27.73)μg·h·mL^(-1)降至(550.39±12.34)μg·h·mL^(-1),TSN的AUC由(1.08±0.72)μg·h·mL^(-1)降至(0.65±0.04)μg·h·mL^(-1),GA的AUC由(43.64±3.10)μg·h·mL^(-1)增至(96.21±3.75)μg·h·mL^(-1);组织分布结果表明,Suc-GTS-lip组肝脏中Sal B的AUC和C_(max)分别是GTS-lip组的10.21、4.44倍,Suc-GTS-lip组中Sal B、TSN、GA对肝脏的靶向效率分别为40.66%、3.06%、22.08%;体内成像研究显示,修饰后的脂质体有肝脏积聚的趋势;MTT实验结果显示,Suc-GTS-lip能显著抑制HSC的增殖;RT-PCR结果显示,各给药组MMP-1的表达均显著增加;各给药组TIMP-1与TIMP-2表达显著降低;各给药组Ⅰ型胶原(collagen-Ⅰ)与Ⅲ型胶原(collagen-Ⅲ)的mRNA表达显著降低。实验结果表明,Suc-GTS-lip具有肝脏靶向性,且能够抑制HSC增殖和诱导其凋亡,为Suc-GTS-lip靶向治疗肝纤维化提供了实验依据。

The 3-succinate-30-stearyl glycyrrhetinic acid(18-GA-Suc) was inserted into glycyrrhetinic acid(GA)-tanshinone Ⅱ_A(TSN)-salvianolic acid B(Sal B) liposome(GTS-lip) to prepare liver targeting compound liposome(Suc-GTS-lip) mediated by GA receptors.Next,pharmacokinetics and tissue distribution of Suc-GTS-lip and GTS-lip were compared by UPLC,and in vivo imaging tracking of Suc-GTS-lip was conducted.The authors investigated the effect of Suc-GTS-lip on the proliferation inhibition of hepatic stellate cells(HSC) and explored their molecular mechanism of improving liver fibrosis.Pharmacokinetic results showed that the AUC_(Sal B) decreased from(636.06±27.73) μg·h·mL^(-1) to(550.39±12.34) μg·h·mL^(-1),and the AUC_(TSN) decreased from(1.08±0.72) μg·h·mL^(-1) to(0.65±0.04) μg·h·mL^(-1),but the AUC_(GA) increased from(43.64±3.10) μg·h·mL^(-1) to(96.21±3.75) μg·h·mL^(-1).The results of tissue distribution showed that the AUC_(Sal B) and C_(max) of Sal B in the liver of the Suc-GTS-lip group were 10.21 and 4.44 times those of the GTS-lip group,respectively.The liver targeting efficiency of Sal B,TSN,and GA in the Suc-GTS-lip group was 40.66%,3.06%,and 22.08%,respectively.In vivo imaging studies showed that the modified liposomes tended to accumulate in the liver.MTT results showed that Suc-GTS-lip could significantly inhibit the proliferation of HSC,and RT-PCR results showed that the expression of MMP-1 was significantly increased in all groups,but that of TIMP-1 and TIMP-2 was significantly decreased.The mRNA expressions of collagen-I and collagen-Ⅲ were significantly decreased in all groups.The experimental results showed that Suc-GTS-lip had liver targeting,and it could inhibit the proliferation of HSC and induce their apoptosis,which provided the experimental basis for the targeted treatment of liver fibrosis by Suc-GTS-lip.