【目的】 观察黄连素调节Hedgehog信号通路对转化生长因子β1（TGF-β1）诱导的肝星状细胞增殖和活化的影响。【方法】 培养HSC-T6细胞，分为空白组，TGF-β1组，黄连素低、中、高剂量组及黄连素高剂量+ purmorphamine（Hedgehog信号通路 激活剂）组。分组处理后，四甲基偶氮唑盐（MTT）法检测 HSC-T6细胞增殖能力，流式细胞术检测细胞凋亡，免疫荧光染色 法检测细胞Ⅰ型胶原（CollagenⅠ）、α-平滑肌肌动蛋白（α-SMA）表达变化，实时荧光定量聚合酶链式反应（qRT-PCR）法检测 细胞增殖细胞核抗原（PCNA）、Bcl-2、Bcl-2 相关 X 蛋白（Bax）mRNA 表达水平，蛋白免疫印迹（Western Blot）法检测细胞 Hedgehog信号通路相关因子Smo、Gli-1、Shh、Ptch蛋白表达水平。【结果】 与空白组比较，TGF-β1组HSC-T6细胞增殖率， CollagenⅠ、α-SMA阳性蛋白表达，PCNA mRNA、Bcl-2 mRNA表达水平及Smo、Gli-1、Shh、Ptch蛋白表达水平显著升高， 凋亡率及 Bax mRNA 表达水平显著降低（均 P＜0.05）；与 TGF-β1 组比较，黄连素低、中、高剂量组 HSC-T6 细胞增殖率， CollagenⅠ、α-SMA阳性蛋白表达，PCNA mRNA、Bcl-2 mRNA表达水平及Smo、Gli-1、Shh、Ptch蛋白表达水平显著降低， 凋 亡 率 及 Bax mRNA 表 达 水 平 显 著 升 高（均 P＜0.05）， 呈 剂 量 依 赖 性 ； 与 黄 连 素 高 剂 量 组 比 较 ， 黄 连 素 高 剂 量 +purmorphamine 组 HSC-T6 细胞增殖率，CollagenⅠ、α-SMA 阳性蛋白表达，PCNA mRNA、Bcl-2 mRNA 表达水平及 Smo、 Gli-1、Shh、Ptch 蛋白表达水平显著升高，凋亡率及 Bax mRNA 表达水平显著降低（均 P＜0.05）。【结论】 黄连素通过调节 Hedgehog信号通路促进TGF-β1诱导肝星状细胞凋亡，抑制其增殖和活化。

Objective To observe the effect of berberine on the proliferation and activation of hepatic stellate cells induced by transforming growth factor β1（TGF-β1）via regulating Hedgehog signaling pathway. Methods HSCT6 cells were cultured and divided into blank group，TGF-β1 group，berberine low-，medium- and high- dose groups，berberine high-dose+purmorphamine（Hedgehog signaling pathway activator）group. After treatment，the proliferation of HSC-T6 cells was detected by methyl thiazolyl tetrazolium（MTT）assay. The apoptosis of HSC-T6 cells was detected by flow cytometry. The expressions of collagen I and α -smooth muscle actin（α -SMA）were detected by immunofluorescence staining. The mRNA expression levels of proliferating cell nuclear antigen （PCNA）， Bcl-2 and Bcl-2 associated X protein（Bax）were detected by real-time fluorescence quantitative polymerase chain reaction（qRT-PCR）. The protein expression levels of Hedgehog signaling pathway related factors Smo， Gli-1， Shh and Ptch were detected by Western Blot. Results Compared with the blank group， the proliferation rate of HSC-T6 cells， the positive protein expressions of Collagen Ⅰ and α -SMA， the mRNA expression levels of PCNA and Bcl-2，and the protein expression levels of Smo，Gli-1，Shh and Ptch in the TGF-β1 group were significantly increased，while the apoptosis rate and the mRNA expression level of Bax were significantly decreased（all P＜0.05）. Compared with TGF-β1 group，the proliferation rate of HSC-T6 cells，the expression of Collagen I and α-SMA positive protein，the mRNA expression levels of PCNA and Bcl-2，and the protein expression levels of Smo，Gli-1，Shh and Ptch in berberine low-，medium- and high- dose groups were significantly decreased，while the apoptosis rate and the mRNA expression level of Bax were significantly increased （P＜0.05）in a dose-dependent manner. Compared with the high-dose berberine group，the proliferation rate of HSC-T6 cells，the positive proteins expression of Collagen Ⅰ and α-SMA，the mRNA expression levels of PCNA and Bcl-2， and the protein expression levels of Smo， Gli-1， Shh and Ptch in the high-dose berberine+ purmorphamine group were significantly increased，while the apoptosis rate and the mRNA expression level of Bax were significantly decreased（all P＜0.05）. Conclusion Berberine promotes TGF-β1-induced apoptosis of hepatic stellate cells and inhibits their proliferation and activation by regulating Hedgehog signaling pathway.

R285.5

河南省医学科技攻关计划联合共建项目（编号：LHGJ20200680）