内毒素对大鼠肝星状细胞活化和瘦素表达的影响

Influence of endotoxin on activation and leptin expression of rat hepatic stellate cells

目的观察内毒素对肝星状细胞(HSC)增殖、Ⅰ型胶原及瘦素表达的影响。方法取传代培养的大鼠肝星状细胞系rHSC-99,以1×108L-1浓度接种于96孔培养板,每孔100μL,加入内毒素使内毒素浓度分别为0.1,1.0,10.0 EIU/mL,同时设对照组,以MTT比色法检测培养24 h和48 h时HSC吸光度。采用ELISA法检测其培养48 h时瘦素和I型胶原水平。结果内毒素组浓度为0.1,1.0 EIU/mL时HSC的平均每孔吸光度和瘦素浓度均显著高于对照组(P<0.05)。内毒素组浓度为1.0 EIU/mL时HSC的平均I型胶原水平显著高于对照组。而内毒素浓度为10.0 EIU/mL时HSC的胶原和瘦素表达水平无明显增高。结论一定浓度的内毒素可以促进大鼠HSC活化,使细胞增殖和I型胶原的表达增加,并促进HSC瘦素的表达,这可能是引起肝纤维化的重要因素。

Objective It is to observe the influence of endotoxin on hepatic stellate cell （HSC） proliferation, type Ⅰ collagen and leptin expression. Methods The HSCs （rHSC - 99） at a content of 1×10^8L^-1 were inoculated into 96 - hole plate for 100 μL per hole, and endotoxin was added to make the concentration 0.1, 1.0 and 10.0 EIU/mL respectively. Control group was established synchronously. The absorbance of HSC was measured with MTT colorimetric method after cultivation for 24 h and 48 h, and the levels of leptin and type Ⅰ collagen were measured with ELISA method after cultivation for 48 h. Results The mean absorbance per hole and leptin content of HSC with the concentration 0.1 and 1.0 EIU/mL in endotoxin group were both significantly higher than those in control group （P〈 0.05）. The mean level of type Ⅰ collagen of HSC with the concentration 1.0 EIU/mL in endotoxin group was significantly higher than that in control group. The levels of collagen and leptin expression didn＇t rise obviously with the concentration 10.0 EIU/mL in endotoxin group. Conclusion Endotoxin with definite concentration can promote HSC activation, make the cell proliferation, increase type Ⅰ collagen expression and promote leptin expression, which may be the important factor to induce hepatic fibrosis.