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血管紧张素II对大鼠肝星状细胞收缩中Rho-Rock信号通路的影响

[Effect of angiotensin II on Rho-Rock pathway in rat hepatic stellate cell contraction].

作者信息

Zhang Xiao-Lan, Li Xu, Xiao Bing, Huang Mao-Liang, Meng Ying, Li Ying-Fei, Wang Yuan-Yuan, Song Wei-Bing

机构信息

Department of Digestive Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2008 Jun;28(6):968-71.

Abstract

OBJECTIVE

To investigate the mechanisms of angiotonin II (AngII)-induced Ca(2+)-independent pathways mediated by Rho kinase in hepatic stellate cells (HSCs).

METHODS

HSC-T6 cells were treated with 1 micromol/L of AngII, and the subsequent cell contraction was directly observed with silicone rubber membrane culture method. The cells with 10 micromol/L AngII treatment were examined for myosin light chain (MLC) phosphorylation level using Western blotting, and the effects of irbesartan (a specific inhibitor of AngII 1- receptor) and Y27632 (a Rho kinase inhibitor) on AngII-induced MLC phosphorylation were evaluated. RT-PCR was used to detect the expression of Rock2 in Ca(2+)- independent pathways mediated by Rho kinase.

RESULTS

AngII induced HSC contraction and time-dependent MLC phosphorylation changes, which peaked 15 min after the treatment followed by gradual reduction. Irbesartan or Y27632 treatment significantly lowered MLC phosphorylation level in AngII-induced cells (P<0.01). The mRNA expression of Rock2 increased significantly after AngII treatment (P<0.01), but decreased following subsequent irbesartan or Y27632 treatment.

CONCLUSION

AngII induces HSC contraction through Ca(2+)-independent pathways mediated by Rho kinase.

摘要

目的

探讨血管紧张素II(AngII)通过Rho激酶介导的肝星状细胞(HSCs)中不依赖钙离子的途径的机制。

方法

用1微摩尔/升的AngII处理HSC-T6细胞,采用硅橡胶膜培养法直接观察随后的细胞收缩情况。用蛋白质印迹法检测用10微摩尔/升AngII处理的细胞的肌球蛋白轻链(MLC)磷酸化水平,并评估厄贝沙坦(一种AngII 1受体特异性抑制剂)和Y27632(一种Rho激酶抑制剂)对AngII诱导的MLC磷酸化的影响。采用逆转录-聚合酶链反应(RT-PCR)检测Rho激酶介导的不依赖钙离子途径中Rock2的表达。

结果

AngII诱导HSC收缩及MLC磷酸化水平随时间的变化,处理后15分钟达到峰值,随后逐渐降低。厄贝沙坦或Y27632处理显著降低了AngII诱导细胞中的MLC磷酸化水平(P<0.01)。AngII处理后Rock2的mRNA表达显著增加(P<0.01),但随后用厄贝沙坦或Y27632处理后表达下降。

结论

AngII通过Rho激酶介导的不依赖钙离子的途径诱导HSC收缩。

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