醉鱼草苷Ⅳ对大鼠肝星状细胞增殖、活化的影响

Effects of Buddlejasaponin Ⅳ on Proliferation and Activation of Hepatic Stellate Cells

目的:观察醉鱼草苷Ⅳ对大鼠贮脂细胞株(HSC-T6)增殖及活化的影响。方法:用不同质量浓度的醉鱼草苷Ⅳ处理HSC-T6,采用噻唑蓝(MTT)法测定醉鱼草苷Ⅳ对HSC-T6增殖的影响。采用20,14,9.8,6.86 mg.L-1醉鱼草苷Ⅳ干预细胞24 h,逆转录聚合酶链式反应(RT-PCR)法检测Ⅰ型胶原(Col-Ⅰ),Ⅲ型胶原(Col-Ⅲ)mRNA的表达;Annexin V/PI双染色法结合流式细胞术检测醉鱼草苷Ⅳ对HSC-T6凋亡的影响。结果:醉鱼草苷Ⅳ可显著抑制HSC-T6的增殖,且呈明显的量效关系。醉鱼草苷Ⅳ(6.86 mg.L-1)对HSC-T6 Col-ⅢmRNA表达具有显著的抑制作用(P<0.05);醉鱼草苷Ⅳ(9.8 mg.L-1)对细胞分泌羟脯氨酸(Hyp)和HSC-T6 Col-ⅠmRNA表达具有显著的抑制作用(P<0.05,P<0.01)及对HSC-T6具有显著的诱导凋亡作用(P<0.01),从而达到抗肝纤维化的作用。结论:醉鱼草苷Ⅳ可体外抑制HSC-T6的增殖和活化,对肝纤维化有潜在的干预作用,其机制可能与抑制贮脂细胞增殖及细胞外基质的合成有关。

Objective： To investigate the effect of buddlejasaponin Ⅳ on proliferation and activation of rat hepatic stellate cell line HSC-T6.Method： HSC-T6 cells was incubated with buddlejasaponin Ⅳ at different concentrations（20,14,9.8,6.86 mg·L-1） for 24 hours.Effect of buddlejasaponin Ⅳ on proliferation of HSC-T6 was observed by MTT assay.Col-I and Col-Ⅲ mRNA expressions were assayed by reverse transcription ploymerase chain reaction（RT-PCR） after incubation.Apoptosis of HSC-T6 was detected by Annexin V/ PI staining combined with flow cytometry method.Result： Buddlejasaponin Ⅳ could inhibit HSC-T6 cell proliferation,in a dose-and time-dependent manner.Buddlejasaponin Ⅳ with 6.86 mg·L-1 inhibited Col-Ⅲ mRNA expression（P0.05）.Buddlejasaponin Ⅳ with 9.8 mg·L-1 decreased Hyp secretion,inhibited Col-I mRNA expression（P0.05） and induced apoptosis significantly（P0.01）,so as to achieve the effect of anti-hepatic fibrosis.Conclusion： Buddlejasaponin Ⅳcould interfere liver fibrosis via inhibiting proliferation and activation of HSC-T6 cells and inhibiting the synthesis of extracellular matrix.