丹参注射液对血管紧张素Ⅱ致肾小球系膜细胞JAK_2/STAT信号通路活化的干预作用研究

Intervention Effect of Danshen Injection(丹参注射液) on JAK/STAT Signal Pathway in Mesangial Cells Activated by Angiotensin Ⅱ

目的:探讨丹参注射液对血管紧张素Ⅱ(AngiotensinⅡ,AngⅡ)致肾小球系膜细胞Janus激酶/信号转导子与转录激活子(Janus kinase/signal ransducers and activators of transcription,JAK/STAT)活化的干预。方法:采用不同浓度AngⅡ与系膜细胞共孵育,采用AngⅡ受体阻断剂氯沙坦和不同浓度丹参注射液阻断。用Western Blot方法检测细胞Rac-1、P-JAK2、P-STAT1、P-STAT3水平;采用ELISA方法测定细胞上清液中纤维连接蛋白(fibronectin,FN)的含量。结果:AngⅡ致系膜细胞JAK/STAT信号通路中P-STAT1、P-STAT3、P-JAK1、Rac-1表达增加,同时使系膜细胞分泌FN水平升高,并呈剂量依赖性,在浓度为10-6mmol/L达到相对稳定状态,10-5mmol/L时达到最高。加用丹参注射液后使升高的Rac-1、P-JAK2、P-STAT1、P-STAT3表达以及FN水平恢复到正常水平。结论:丹参注射液能够通过抑制AngⅡ诱导的系膜细胞JAK/STAT信号途径的活化,从而减少FN的表达,发挥抗肾小球硬化作用。

Objective：To investigate the intervention effect of Danshen Injection on the activation of JAK/ STAT signal pathway in mesangial cells induced by angiotensin H ~ Methods： Quiescent rat＇ s mesangial cells were cultured by different concentrations of Ang II or by combination with the same concentration Ang Ⅱ with Dan- shen Injection or Losartan injection. The protein of P - STAT1 , P - STAT3, P - JAK1 and Rae - 1 were measured by Western blotting,ELISA was used to detect the expression of fibronectin（FN） in free MEM medium. Results： In cultured mesangial cells, Ang II increased expression of Rac - 1 and phosphorylation of JAK2 , STATl , STAT3 in the form of a dose - dependent mechanism as well as the level of fibronectin in the conditioned medium,and the maximal induction of those was observed at the highest dose of 10-s mmol/L Ang Ⅱ,10-6 mmol/L AngU was the metastable state. Danshen Injection could notably attenuated expressions of Rac - 1 and phosphorylation of JAK2, STAT1 and STATz induced by Ang I[ in a concentration - dependent manner. Meanwhile,it suppressed the production of FN to normal level. Conclusion： Danshen Injection can decrease the phosphorylation of JAK/STAT in mesangial cells ,which due to its ability of inhibiting the activities of angiotensin II. Danshen Injection might be an effective therapeutic strategy for glomerular sclerosis.