吲哚-3-原醇对乙醛所致精密肝切片中星状细胞活化的影响

目的利用精密肝切片(PCLS)技术,研究吲哚-3-原醇(I3C)对乙醛活化肝星状细胞(HSCs)的作用及其机制。方法将200～800μmol.L-1的I3C及700μmol.L-1乙醛与PCLS共同孵育6h,免疫组化法分析肝切片中α-平滑肌肌动蛋白(α-SMA)表达,并检测培养液中谷胱甘肽S-转移酶(GST)和乳酸脱氢酶(LDH)活性、转化生长因子-β1(TGF-β1)含量、基质金属蛋白酶-1(MMP-1)及基质金属蛋白酶抑制物-1(TIMP-1)表达量及组织丙二醛(MDA)和羟脯氨酸(Hyp)含量。结果200～800μmol.L-1I3C可不同程度的减少乙醛激活的HSCs,并可明显降低乙醛升高的培养液中GST、LDH活性和肝组织中MDA和Hyp含量(P<0.05或P<0.01),呈良好的浓度依赖性。I3C给药组与乙醛对照组比较,培养液中TGF-β1含量降低(P<0.01),MMP-1/TIMP-1蛋白表达比值升高(P<0.01)。结论I3C能有效拮抗乙醛所致的HSCs活化,其机制与降低细胞氧化应激和促进基质胶原降解有关。

Effects of indole-3-carbinol on hepatic stellate cells activated by acetaldehyde in precision-cut liver slices

Aim To investigate effects of indole-3-carbinol (I3C) on hepatic stellate cells (HSCs) activated by acetaldehyde in precision-cut liver slices (PCLS). Methods PCLS were incubated with 700 μmol·L-1 acetaldehyde and 200 ~ 800 μmol·L-1 I3C for 6 h. The expression of α-smooth muscle actin(α-SMA) in liver slices was analyzed by immunohistochemistry. The leakages of glutathione S-transferase (GST), lactate dehydrogenase (LDH) and content of transforming growth factor-β1 (TGF-β1) in media were assayed. Contents of malondialdehyde (MDA) and hydroxyproline (Hyp) in tissue were also determined. Expressions of matrix metalloproteinases-1 (MMP-1) and tissue inhibitor of metalloproteinase (TIMP-1) in media were analyzed by the western blot. Results The increase of activated HSCs due to acetaldehyde was inhibited by I3C (200~800 μmol·L-1). Meanwhile, I3C treatment (200~800 μmol·L-1) showed significant and concentration-dependent antagonistic actions on the increment of GST, LDH leakages into the media and MDA, Hyp contents in tissues induced by acetaldehyde. The increase of TGF-β1 was also remarkable inversed by I3C (200~800 μmol·L-1). As compared with acetaldehyde group, the ratio of MMP-1/TIMP-1 was increased significantly by I3C treatment (800 μmol·L-1)(P<0.01). Conclusions I3C markedly protects liver slices from acetaldehyde-induced HSC activation, and it might involve the mechanism of the antioxidation against acetaldehyde and the promotion of ECM degradation.