蛇葡萄素对大鼠肝星状细胞增殖及胶原蛋白、细胞因子生成的影响

Effect of Ampelopsin on HSC-T6 Cell Proliferation and Content of Collage and Cytokine

摘要:

目的：研究蛇葡萄素对大鼠肝星状细胞（HSC-T6）的增殖及胶原蛋白、细胞因子生成的影响。方法：对HSC-T6增殖的影响：用含10%胎牛血清的高糖DMEM培养液制备成HSC-T6细胞3×10⁴个/mL细胞悬液100 μL/孔接种于96孔培养板。设细胞对照组、秋水仙碱1.0 mg·L^-1组和100.0，75.0，50.0，25.0，12.5 mg·L^-15个不同质量浓度蛇葡萄素组，置37 ℃ 5%CO₂培养48 h。采用MTT法检测蛇葡萄素对大鼠肝细胞HSC-T6增殖的影响。对HSC-T6生成Ⅰ，Ⅲ，Ⅳ型胶原蛋白及细胞因子（TGF-β₁，PDGF）含量的影响：取终质量浓度分别为50.0，25.0，12.5 mg·L^-1蛇葡萄素，3×10⁴个/mL细胞悬液100 μL/孔细胞液接种于96孔培养板。阳性组秋水仙碱的质量浓度为1.0 mg·L^-1。置37 ℃ 5%CO₂培养48 h。ELISA法检测HSC-T6细胞生成的胶原蛋白（Ⅰ，Ⅲ，Ⅳ）、转化生长因子（TGF-β₁）及血小板衍生因子（PDGF）含量。结果：蛇葡萄素100.0，75.0 mg·L^-1对HSC-T6细胞的增殖有明显抑制作用，其无毒浓度为50.0 mg·L^-1。50 mg·L^-1的蛇葡萄素可使HSC-T6细胞生成的Ⅰ，Ⅲ，Ⅳ型胶原蛋白，TGF-β₁，PDGF含量明显低于细胞对照组（P<0.05），25 mg·L^-1的蛇葡萄素也可使HSC-T6细胞生成的Ⅲ型胶原蛋白含量明显低于细胞对照组（P<0.05）。结论：蛇葡萄素能抑制HSC-T6增殖并可使HSC-T6生成的Ⅰ，Ⅲ，Ⅳ型胶原蛋白，TGF-β₁，PDGF含量降低。

Abstract:

Objective: To study the effect of ampelopsin on the HSC-T6 cell proliferation rate and the content of collage and cytokine Method: Effect of ampelopsin on the cell (HSC-T6) proliferation rate. High glucose DMEM medium containing 10% fetal bovine serum was used to prepare HSC-T6 cells 3×10⁴ cells/mL suspension 100 μL/well, which was seeded in 96-well culture plates. The control group, colchicine 1.0 mg·L^-1 group and 100.0, 75.0, 50.0, 25.0, 12.5 mg·L^-1, 5 different concentrations of ampelopsin groups were designed. Cells were cultured at 37 ℃ 5%CO₂ for 48 h. The cell proliferation inhibition rate was measured by MTT。Effect of ampelopsin on the content of collage(Ⅰ, Ⅲ, Ⅳ) and cytokine (TGF-β₁, PDGF) was dected by ELISA. Result: Ampelopsin at 100.0, 75.0 mg·L^-1 might effectively inhibit proliferation of HSC-T6 cells, and the non-toxic drug concentration was 50.0 mg·L^-1, the contents of collage Ⅰ, Ⅲ, Ⅳ and TGF-β₁, PDGF in HSC-T6 cells were significantly lower than control group at 50.0 mg·L^-1 of ampelopsin (P<0.05), the contents of collage Ⅲ in HSC-T6 cells was significantly lower than control group by ampelopsin at 25.0 mg·L^-1(P<0.05). Conclusion: Ampelopsin could inhibit the proliferation of HSC-T6 and reduce the content of collage Ⅰ, Ⅲ, Ⅳ and TGF-β₁, PDGF.