参仙升脉口服液介导TRPC3/TRPC6通路抑制病态窦房结综合征钙离子内流

Shenxian Shengmai Oral Liquid Mediates TRPC3/TRPC6 Pathway to Inhibit Calcium Ion Inflow in Sick Sinus Syndrome

目的探讨参仙升脉口服液对病态窦房结综合征(sick sinus syndrome,SSS)小鼠钙离子内流的影响。方法C57/b6小鼠60只,随机数字表法将小鼠分为病态窦房结综合征模型组(SSS)、参仙升脉口服液治疗组(SSOL)、假手术组(Sham)。利用微量渗透泵(mini osmotic pump)技术泵入血管紧张素Ⅱ建立病态窦房结综合征模型(sick sinus syndrome,SSS),造模后利用生物信号采集系统规律性进行心电图监测(2次/周),观察3组小鼠心电图变化。成模后(30 d,即第31天开始),实验组小鼠每日灌胃参仙升脉口服液0.2 m L,模型组与假手术组每日灌胃同等剂量生理盐水,给药后继续2次/周心电图监测,30 d后取材。HE染色、Masson染色观察窦房结组织病理变化;Western-blot检测TRPC3、TRPC6蛋白表达水平;PCR检测CAV1.2、Cav3.1型Ca^2+mRNA水平。结果模型组小鼠窦房结组织纤维化程度较重,实验组小鼠窦房结组织纤维化程度明显好于模型组。模型组TRPC3、TRPC6的表达明显高于实验组与假手术组,且CAV1.2、Cav3.1的表达同样明显高于实验组与假手术组。结论参仙升脉口服液能介导TRPC3、TRPC6通路抑制Ca+内流,从而延缓窦房结组织纤维化进程。

Objective To investigate the effect of Shenxian Shengmai Oral Liquid on calcium influx in sick sinus syndrome(SSS)mice.Methods Sixty SPF clean-grade mice were enrolled in this study.The model of SSS was established by pumping angiotensin II using Mini osmotic pump technique.The mice were divided into model group,sham operation group and treatment group by random number table method.Mice were subjected to electrocardiogram monitoring(2 times/week)using the biosignal acquisition system after modeling,and the electrocardiogram changes of the three groups were observed.After the model was formed(30 days,starting on the 31 st day),the mice in the treatment group were intragastrically administered with0.2 m L of Shenxian Shengmai Oral Liquid.The model group and the sham operation group were intragastrically administered with the same dose of normal saline daily,2 times/week ECG monitoring.After 30 days,the pathological changes of sinus node were observed by HE staining.The expressions of TRPC3 and TRPC6 were detected by Western-blot.The levels of CAV1.2 and Cav3.1 Ca+mRNA were detected by PCR.Results The degree of fibrosis in the sinoatrial node of the model group was heavier.The degree of fibrosis in the sinoatrial node of the treatment group was significantly better than that of the model group.The expressions of TRPC3 and TRPC6 in the model group were significantly higher than those in the treatment group and the sham operation group,and the expressions of CAV1.2 and Cav3.1 were also significantly higher than those in the treatment group and the sham operation group.Conclusion Shenxian Shengmai Oral Liquid can mediate TRPC3 and TRPC6 pathways to inhibit Ca^2+influx,and thus delaying the process of sinus node tissue fibrosis.