Shenxian Shengmai Oral Liquid Inhibits Calcium Ion Overload Mediated By TRPC3 Pathway And Improves Sinus Node Fibrosis In SSS Mice

Purpose:To clarify the effect of Shenxian Shengmai Oral Liquid on improving sinus node fibrosis in SSS mice,and to explore the mechanism of TRPC3 pathway in improving sinus node fibrosis of SSS mice.Material and method:Thirty C57 / b6 mice were randomly divided into the sick sinus node syndrome model group(SSS),the Shenxian Shengmai oral liquid treatment group(SSOL),and the sham operation group(Sham).Mini osmotic pump technology was used to pump angiotensin ? to establish a sick sinus syndrome(SSS)model.After modeling,the mice in the Shenxian Shengmai Oral Solution group were given 15ml/kg of Shenxian Shengmai Oral Solution daily,and the model group and the sham operation group were given the same daily dose of normal saline,and the biological signal acquisition system was used regularly.ECG monitoring was performed(0 days before modeling,7 days,14 days,21 days,and 28 days after surgery),and the ECG changes of the three groups of mice were observed,and samples were taken 30 days later.HE staining and Masson staining were used to observe the pathological changes of sinoatrial node tissues;WB detected fibrosis-related factor type I collagen ?1 chain(Col1A1),matrix metalloproteinase 2(MMP-2)and transforming growth factor beta(TGF-?),and TRPC3,Cav3.1,Cav3.2,Cav1.2 protein expression;IF observation and analysis of Ca2 + expression in sinoatrial node cells;PCR detection of fibrosis-related factors Col1A1,MMP-2,TGF-? and Cav3.1,Cav3.2,Cav1.2 Ca2+ m RNA levels.Results:1.Heart rate: There was no significant change in heart rate in the sham operation group.The heart rate of mice in each group was within the normal range on day 0.Compared with the sham operation group,the heart rate of the model group showed a downward trend after modeling,and the downward trend was obvious.There was still a downward trend after 28 days of modeling.After the intervention of Shenxian Shengmai Oral Liquid in the experimental group,the heart rate increased slightly compared with the model group.The heart rate increased significantly from the 14 th day,but it was still slower than the sham operation group(P <0.05).2.HE staining showed that the structure of sinoatrial node cells in the sham operation group was complete,with clear boundaries,epithelial cell distribution was seen in the ganglion space,and there was no fibrosis in the sinoatrial node cells.The sinus node in the model group had obvious fibrosis and severe fibrosis.Compared with the model group,the degree of fibrosis in the experimental group was significantly reduced,but compared with the sham operation group,fibrosis was still seen.3.Masson staining showed that the interstitial fibers in the sinoatrial node area of the sham operation group showed a reticular scaffold,and the surface arteries of the sinoatrial node tissue were connected to the fibrous network structure without fibrosis.The tissue cells in the sinoatrial node area of the mouse in the model group Blurred,it can be seen that a large number of bundled connective tissues are proliferated,the fibrous structure of the stent is damaged,and the degree of fibrosis is heavier;the parenchymal cells in the experimental group are arranged neatly and densely,and a small amount of collagen fibrous tissue is seen in the intercellular space,and the degree of fibrosis is light.4.Immunofluorescence: We performed immunofluorescence detection on the sinoatrial node area of the mouse,and found that compared with the Sham group,the Ca + expression in the sinus node area of the SSS group was significantly increased.Ca2+ expression was significantly lower than that in the model group,but still higher than that in the Sham group(P <0.05).5.WB test: We tested the fibrosis-related factors Col1A1,MMP-2,TGF-?,TRPC3,and CAV3.1,CAV3.2,and CAV1.2 proteins in three groups of mice.The results showed that the mice in the SSS group The contents of Col1A1,MMP-2,and TGF-?were all increased,and the contents of TRPC3,Cav3.1,Cav3.2,and Cav1.2 proteins were also highly expressed compared to the Sham group,while the mice in the SSOL group were Col1A1,The contents of MMP-2,TGF-? and the expressions of TRPC3,Cav3.1,Cav3.2,and Cav1.2 proteins were significantly lower than those in the SSS group,but still higher than those in the Sham group.6.q PCR: q PCR further detected the expression of TRPC3,Cav3.1,Cav3.2,and Cav1.2 proteins at the m RNA level,and the results showed that compared with the Sham group,the TRPC3,Cav3.1,Cav3.2,and Cav1.2 in the SSS and SSOL groups The expression of protein was higher than that of Sham group,but the expression of SSOL group was significantly lower than that of SSS group.Conclusion:1.Shenxian Shengmai Oral Liquid can increase heart rate and improve sinus node fibrosis in sick sinus node syndrome mice.2.The TRPC3 pathway plays an important role in improving the sinus node fibrosis of Shenxian Shengmai Oral Liquid,and its mechanism is to inhibit the extracellular calcium ion inflow,thereby delaying the process of sinus node fibrosis.