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醛固酮通过不依赖钙离子的途径刺激肝星状细胞收缩。

[Aldosterone stimulates hepatic stellate cells contraction via Ca2+-independent pathways].

作者信息

Zhang Xiao-lan, Xiao Bing, Meng Ying, Li Xu

机构信息

Department of Senile Disease, Taihe Hospital, Hubei Medical College, Shiyan Hubei Province 442000, China.

出版信息

Zhonghua Gan Zang Bing Za Zhi. 2011 Jul;19(7):537-41. doi: 10.3760/cma.j.issn.1007-3418.2011.07.016.

Abstract

OBJECTIVE

To investigate the mechanisms of Aldosterone stimulating hepatic stellate cells(HSCs) contraction via Ca2+-independent pathways.

METHODS

HSC-T6 cell line was pre-disposed with Aldo 10mumol/L. The cell contraction was detected by silicone-rubber-membrane cultivation directly. The concentration variation of intracellular free calcium in rat HSC was observed by laser confocal microscopy. Besides, HSC-T6 cell line was under pre-disposal treatment with the blocking agents of Aldo receptor -antisterone, protein kinase C (PKC) special blocking agent-Stauro, Rho kinase blocking agent-Y27632 and MLCK special blocking agent-ML-7 respectively prior to stimulation with aldosterone. RT-PCR was used to detect the expression of Rock2, RhoAGTP and RhoGEF in Ca2+- independent pathways mediated by Rho-kinase.

RESULTS

Aldo could induce HSCs contraction. The concentration of intracellular free calcium in rat HSCs had no change after pre-disposal treatment with Aldo. The mRNA expressions of Rock2, RhoAGTP and RhoGEF increased significantly after treatment with Aldo (0.770+/-0.049, 0.960+/-0.096, 0.180+/-0.006, P is less than 0.01).When inhibited with antisterone, the mRNA expressions of the three elements were (0.440+/-0.166, 0.370+/-0.180 and 0.050+/-0.001, P is less than 0.01), lower than that of Aldo group, but higher in ML-7+Stauro + Aldo groups (0.940+/-0.066, 1.330+/-0.192 and 0.160+/-0.007, P is less than 0.05) as compared to the control group (0.140+/-0.023, 0.540+/-0.111 and 0.110+/-0.012). In the Y27632 + ML-7 + Stauro+Aldo group, the mRNA expression of RhoGEF (0.290+/-0.004, P is less than 0.01)was higher than that of the ML-7 + Stauro + Aldo group (0.160+/-0.007).

CONCLUSION

Aldo could induce HSCs contraction via Ca2+-independent pathways and Rho-Rock pathway involved in the process.

摘要

目的

探讨醛固酮通过非钙依赖途径刺激肝星状细胞(HSCs)收缩的机制。

方法

用10μmol/L醛固酮预处理HSC-T6细胞系。通过硅橡胶膜培养直接检测细胞收缩情况。用激光共聚焦显微镜观察大鼠肝星状细胞内游离钙浓度变化。此外,在用醛固酮刺激前,分别用醛固酮受体阻断剂 - 抗醛固酮、蛋白激酶C(PKC)特异性阻断剂 - 星形孢菌素、Rho激酶阻断剂 - Y27632和肌球蛋白轻链激酶(MLCK)特异性阻断剂 - ML-7对HSC-T6细胞系进行预处理。采用RT-PCR检测Rho激酶介导的非钙依赖途径中Rock2、RhoAGTP和RhoGEF的表达。

结果

醛固酮可诱导肝星状细胞收缩。用醛固酮预处理后,大鼠肝星状细胞内游离钙浓度无变化。用醛固酮处理后,Rock2、RhoAGTP和RhoGEF的mRNA表达显著增加(0.770±0.049,0.960±0.096,0.180±0.006,P<0.01)。用抗醛固酮抑制后,这三种成分的mRNA表达分别为(0.440±0.166,0.370±0.180和0.050±0.001,P<0.01),低于醛固酮组,但与对照组(0.140±0.023,0.540±0.111和0.110±0.012)相比,在ML-7 + 星形孢菌素 + 醛固酮组中更高(0.940±0.066,1.330±0.192和0.160±0.007,P<0.05)。在Y27632 + ML-7 + 星形孢菌素 + 醛固酮组中,RhoGEF的mRNA表达(0.290±0.004,P<0.01)高于ML-7 + 星形孢菌素 + 醛固酮组(0.160±0.007)。

结论

醛固酮可通过非钙依赖途径诱导肝星状细胞收缩,且Rho-Rock途径参与了这一过程。

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