Effect Of Simiao Yong’an Decoction On Cardiac Fibroblasts Proliferation And Smad3 Signal Transduction Pathway In Neonatal Rats

In this study,we established the Ang II-induced proliferation of cardiac fibroblasts（CFs）and used different concentrations of Simiao Yong’an Decoction to observe the effects of Ang II on CFs proliferation and TGF-β1/Smad3 signal transduction.To investigate the effect of SJM on the proliferation of CFs and its regulatory mechanism.Neonatal rat CFs were isolated and cultured and identified.Ang II(10^-66 mol·L^-1)was used to establish the CFs proliferation model.Different concentration of Simiao Yong’an Decoction alcohol precipitated solution medicine was used for CFs proliferation intervention.The cultivated CFs were divided into five groups:A,B,C,D,E according to experimental requirements.That is,group A:alcohol precipitated solution without Ang II and Simiao Yong’an Decoction;group B:Ang II-only treatment,and alcohol precipitated solution medicine without Simiao Yong’an Decoction;group C:treatment with Ang II treatment,and add 30%Simiao Yong’an Decoction alcohol precipitated solution medicine to intervene;group D:Add Ang II treatment,and add 20%Simiao Yong’an Decoction alcohol precipitated solution medicine to intervene;group E:Add Ang II treatment,and alcohol precipitated solution medicine intervention with 10%Simiao Yong’an Decoction.MTT colorimetric assay was used to detect the proliferation of CFs in each group.The expression of TGF-β1 in each group of cells was detected by ELISA.The activation level of Smad3 in each group was detected by Western blot to observe the newborn induced by Ang II.The effects of CFs proliferation and TGF-β1/Smad3 signal transduction pathway in rats.The viability of CFs in each group was detected by MTT colorimetry:the OD value in group A was（0.341±0.012）,the OD value in group B was（0.531±0.016）,the OD value in group C was（0.465±0.045）,and the OD value in group D was（0.488±0.020）.The OD value in group E was（0.492±0.017）.Compared with group A,the OD value was significantly increased,indicating that the number of CFs living cells in group B increased significantly,that is,Ang II induced cardiac fibroblast proliferation model successfully.Compared with group B,group E,group C,group D and group B had significantly lower OD values,indicating that the four groups of SJM can reduce the OD value of each well of Ang II induced proliferation model.Decreased proliferation of cardiac fibroblasts induced by Ang II.After one-way analysis of variance,the difference between group B and group A was statistically significant（P<0.05）,with statistical significance;group C,group D,group E,and group B.The difference was statistically significant（P<0.05）.It was also statistically significant.Compared with E group,the OD value was significantly lower in group C.This indicated that the difference in the number of CFs between group C and group E was significant.After one-way analysis of variance,the two groups were compared.There was a significant difference（P<0.05）with statistical significance.The expression of TGF-β1 in CFs of each group was measured by ELISA:the content of TGF-β1 in group A was（87.68±3.93）pg/ml,the content of TGF-β1 in group B was（122.99±4.86）pg/ml,and the content of TGF-β1 in group C.（101.72±4.74）pg/ml,the TGF-β1 content in group D was（108.73±8.28）pg/ml,and the TGF-β1 content in group E was（114.33±7.86）pg/ml.The content of TGF-β1 in group B was higher than that in group A,but the levels of TGF-β1 in groups E,C,and D were lower than those in group B.After one-way analysis of variance,group A and group B were statistically significant.（P<0.05）;The differences between group C,group D,group E and group B were statistically significant（P<0.05）.The levels of TGF-β1 in group C were lower than those in groups E and D.One-way analysis of variance showed significant difference（P<0.05）,with statistical significance.There was no significant difference between E group and D group（P>0.05）.Western blot analysis Smad3 expression level of each group CFs where:A group Smad3/GAPDH ratio（0.301±0.021）,group B Smad3/GAPDH ratio（0.804±0.030）,group C Smad3/GAPDH ratio（0.522±0.014）The ratio of Smad3/GAPDH in group D was（0.534±0.013）,and the ratio of Smad3/GAPDH in group E was（0.547±0.064）.Elevated group B Smad3/GAPDH ratios than in group A,group B Smad3 expression level described above A group,i.e.,successful modeling,group C,D,E group were lower than that in group B Smad3/GAPDH ratio,the Simiao Yong’an Decoction of each group can be reduced Smad3 expression,ANOVA analysis,group B and group A,a difference is significant,statistically significant（P<0.05）;group C,group D,E and group B with The difference was statistically significant（P<0.05）.There was no significant difference between C,D,and E groups（P>0.05）.In summary:Ang II(10^-66 mol·L^-1)can induce CFs proliferation.In the Ang II-induced CFs proliferation model,the addition of Simiao Yong’an Decoction can inhibit the proliferation of CFs after intervention.In this experiment Simiao Yong’an Decoction CFs proliferation inhibition with increased concentration gradient set enhanced,i.e.a concentration of 30%proliferation inhibition CFs strongest,but whether there is a concentration further study provide optimal pharmaceutical efficacy.In addition,it was confirmed that Simiao Yong’an Decoction can reduce the expression of TGF-β1 and Smad3,and its mechanism may inhibit CFs proliferation by regulating TGF-β1/Smad3 signaling pathway.