Notch3 调控肝星状细胞的激活。
Notch3 regulates the activation of hepatic stellate cells.
机构信息
Department of Hepatology and Infectious Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China.
出版信息
World J Gastroenterol. 2012 Mar 28;18(12):1397-403. doi: 10.3748/wjg.v18.i12.1397.
AIM
To investigate whether Notch signaling is involved in liver fibrosis by regulating the activation of hepatic stellate cells (HSCs).
METHODS
Immunohistochemistry was used to detect the expression of Notch3 in fibrotic liver tissues of patients with chronic active hepatitis. The expression of Notch3 in HSC-T6 cells treated or not with transforming growth factor (TGF)-β1 was analyzed by immunofluorescence staining. The expression of Notch3 and myofibroblastic marker α-smooth muscle actin (α-SMA) and collagen I in HSC-T6 cells transfected with pcDNA3.1-N3ICD or control vector were detected by Western blotting and immunofluorescence staining. Moreover, effects of Notch3 knockdown in HSC-T6 by Notch3 siRNA were investigated by Western blotting and immunofluorescence staining.
RESULTS
The expression of Notch3 was significantly up-regulated in fibrotic liver tissues of patients with chronic active hepatitis, but not detected in normal liver tissues. Active Notch signaling was found in HSC-T6 cells. TGF-β1 treatment led to up-regulation of Notch3 expression in HSC-T6 cells, and over-expression of Notch3 increased the expression of α-SMA and collagen I in HSC-T6 without TGF-β1 treatment. Interestingly, transient knockdown of Notch3 decreased the expression of myofibroblastic marker and antagonized TGF-β1-induced expression of α-SMA and collagen I in HSC-T6.
CONCLUSION
Notch3 may regulate the activation of HSCs, and the selective interruption of Notch3 may provide an anti-fibrotic strategy in hepatic fibrosis.
目的
通过调控肝星状细胞（HSCs）的激活来研究 Notch 信号是否参与肝纤维化。
方法
采用免疫组织化学方法检测慢性活动性肝炎患者肝纤维化组织中 Notch3 的表达，并用转化生长因子（TGF）-β1 处理或不处理 HSC-T6 细胞，通过免疫荧光染色分析 Notch3 的表达。用 Western blot 和免疫荧光染色检测 pcDNA3.1-N3ICD 或对照载体转染的 HSC-T6 细胞中 Notch3 和肌成纤维细胞标志物α-平滑肌肌动蛋白（α-SMA）和胶原 I 的表达。此外，通过 Notch3 siRNA 敲低 HSC-T6 中的 Notch3 ，通过 Western blot 和免疫荧光染色检测 Notch3 的作用。
结果
慢性活动性肝炎患者肝纤维化组织中 Notch3 的表达明显上调，而在正常肝组织中未检测到。在 HSC-T6 细胞中发现了活跃的 Notch 信号。TGF-β1 处理导致 HSC-T6 细胞中 Notch3 表达上调，而 Notch3 的过表达增加了无 TGF-β1 处理时 HSC-T6 中 α-SMA 和胶原 I 的表达。有趣的是，瞬时敲低 Notch3 降低了肌成纤维标志物的表达，并拮抗了 TGF-β1 诱导的 HSC-T6 中 α-SMA 和胶原 I 的表达。
结论
Notch3 可能调节 HSCs 的激活，选择性阻断 Notch3 可能为肝纤维化提供一种抗纤维化策略。
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