石蒜碱通过PI3K/AKT信号通路调节自噬抑制HBV诱发肝纤维化

目的 观察石蒜碱抑制乙型肝炎病毒(HBV)诱发肝纤维化的机制.方法 50只雄性C57BL/6小鼠随机分为5组,每组10只:正常对照组、模型组、石蒜碱低、高剂量组(50、100 mg/kg)、阳性对照组(秋水仙碱0.12 mg/kg).除了正常对照组,其余各组采用HBV诱发建立肝纤维化动物模型,治疗组小鼠腹腔注射给药,正...

Lycoline regulates autophagy through PI3K/AKT signaling pathway and inhibits HBV-induced liver fibrosis

Objective To observe the mechanism of lycoline in inhibiting HBV-induced liver fibrosis. Methods Fifty male C57BL/6 mice were randomly divided into five groups, with 10 in each group: normal control group, model group, low dose group of lycorine (50 mg/kg), high dose group of lycorine (100 mg/kg), positive control group (colchicine, 0.12 mg/kg). Except for the normal group, other groups established animal models of liver fibrosis induced by HBV. The mice in the treatment group were injected intraperitoneally, and the mice in the normal control group were injected intraperitoneally with an equal volume of normal saline, once a day for 8 weeks. HSC-T6 with a non-contact co-culture system between cells was cultured. The method of qRT-PCR was used to detect α-SMA, collagen I, and TGF-β1 mRNA levels. Western blot was used to detect related protein expression levels of cell fibrosis, autophagy, PI3K/AKT signaling pathway. PI3K-siRNA and autophagy inhibitor (3-MA) were used to explore the effect of lycoline on liver fibrosis through autophagy. Results In vivo experiments showed that sophoracarpine could reduce liver lobular collagen fibers, and reduce ALT, AST, ALP, hydroxyproline, α-SMA, collagen I, TGF-β1, decrease p-AKT/AKT, p- PI3K/PI3K levels (t=20.937, P=0.001; t=23.505, P=0.001; t=15.778, P=0.007; t=26.279, P=0.001), and increase Beclin-1, LC3 II/LC3 I levels (t=8.050, P=0.008; t=12.593, P=0.003；t=5.095, P=0.008; t=7.202, P=0.004). In vitro experiments showed that 0.313 to 20 μmol/L lycoline could inhibit the proliferation of HSC-T6 cells (P<0.05). Compared with the control group, 2.5 and 10 μmol/L lycoline could reduce α-SMA and collagen I, TGF-β1, decrease p-AKT/AKT, p-PI3K/PI3K levels (t=8.229, P=0.001; t=11.325, P=0.003), and increase LC3 II/LC3 I levels (t=3.798, P= 0.019; t=5.735, P=0.005). Compared with lycoline group, collagen I, -PI3K/PI3K levels in lycoline + PI3K-siRNA group significantly decreased (t=6.877, P=0.004; t=7.206, P=0.002), LC3 II/LC3 I levels were significantly increased (t=4.461, P=0.011), and collagen I, p-PI3K/PI3K levels significantly increased (t=4.073, P=0.015; t=3.105, P=0.036), LC3 II/LC3 I levels significantly down in sophoracarpine +3-MA group (P<0.05) (t=3.454, P=0.026). Conclusion Lycoline can promote hepatocyte autophagy and down-regulate the level of cell fibrosis. The mechanism may be related to the regulation of PI3K/AKT signaling pathway.