熊果苷调控ROCK/JNK通路对自发性高血压大鼠主动脉氧化应激及纤维化的作用研究

Effect of arbutin regulating ROCK/JNK pathway on aorta of spontaneously hypertensive rats

目的研究熊果苷(AR)减轻自发性高血压大鼠(SHR)主动脉氧化应激及纤维化损伤及其对血管ROCK/JNK信号的调控。方法2021年12月,于北部战区总医院心血管外科实验室将10只雄性SHR随机数字表法分为2组:高血压模型组(SHR组,5只)与AR治疗组(AR+SHR组,5只),同周龄健康雄性Wistar大鼠作为正常对照组(Sham组,5只)。AR+SHR组灌胃的方式给予AR 100 mg/(kg·d)4周。采用智能无创血压计测量鼠尾动脉血压;ELISA检测血清中氧化应激丙二醛(MDA)和超氧化物歧化酶(SOD)的水平;HE染色法检测主动脉损伤程度,Masson及天狼星红染色检测血管肌纤维化程度;Western blot法检测主动脉组织CollagenⅠ、CollagenⅢ、RhoA、ROCK1和JNK蛋白的表达水平。结果切片染色结果提示,Sham组细胞结构排列整齐,细胞核完整,肌纤维无断裂;纤维化程度轻;SHR组细胞排解结构紊乱,肌纤维断裂,细胞核溢出,纤维化程度重;SHR+AR组主动脉组织结构有所恢复,肌纤维排列结构得到改善,纤维化程度减轻。与SHR组相比,SHR+AR组大鼠血压和血清中MDA水平,以及动脉组织中CollagenⅠ、RhoA、ROCK1和JNK蛋白表达水平显著降低(P<0.05),血清中SOD水平有明显升高。结论AR治疗可显著减轻SHR平滑肌细胞氧化应激损伤。同时,AR治疗可显著下调ROCK/JNK信号的调控。

Objective To investigate the effects of arbutin(AR)on oxidative stress and fibrosis of aorta in SHR and its regulation on vascular through ROCK/JNK signal.Methods A total of 10 male SHR rats,at the laboratory of Cardiovascular Surgery,General Hospital of the Northern Theater,were randomly divided into two groups:the hypertension model group(SHR group,n=5)and arbutin treatment group(AR+SHR group,n=5)in December 2021.The male healthy Wistar rats at the same age were used as the control group(Sham group,n=5).AR+SHR group was given AR[100 mg/(kg·d)]treatment by gavage for 28 days.The blood pressure of tail artery was measured by intelligent noninvasive sphygmomanometer.The malondialdehyde(MDA)and superoxide dismutase(SOD)in serum were detected by ELISA.The degree of aortic injury was detected by HE staining and the degree of vascular muscle fibrosis was detected by Masson and Sirius red staining.The expression of CollagenⅠ,CollagenⅢ,RhoA,ROCK1 and JNK proteins were detected by Western blot.Results The results of HE staining showed that the cell structure of the Sham group was orderly arranged and the nucleus were complete while the muscle fibers were not broken and the degree of fibrosis was light.In the SHR group,the structure of cells was disordered and the muscle fibers were broken while the nuclei overflowed and the degree of fibrosis was serious.In the SHR+AR group,the aortic tissue structure was restored and the arrangement of muscle fibers was improved while the degree of fibrosis was reduced.Compared with the SHR group,the blood pressure and serum level of MDA in SHR+AR group,as well as the expression level of CollagenⅠ,RhoA,ROCK1 and JNK protein in arterial tissue were significantly decreased(all P<0.05),and the serum level of SOD was significantly increased(P<0.05).Conclusion AR treatment could significantly reduce the oxidative stress injury of SHR smooth muscle cells.At the same time,AR treatment could significantly down regulate ROCK/JNK signal.