The Effect Of Banxia Xiexin Decoction On The Maintenance Of ICC Phenotype And Calcium Regulation Protein

[Objective]In order to explore the mechanism of Banxia Xiexin Tang in regulating gastrointestinal motility disorders,this paper firstly used CiteSpace software to visualize the ICC phenotype related literature in WOS database,and to maintain the ICC phenotype and related ions in Banxia Xiexin Tang.Experimental studies of the effects of channel proteins provide theoretical support.Taking the ICC phenotype as the core,from the perspective of ICC phenotype maintenance and related ion channel proteins,revealing the cellular and molecular mechanisms of Banxia Xiexin Tang affecting the ICC cel phenotype to maintain gastrointestinal motility,The connotation is revealed.[Methods]The serum containing Banxia Xiexin Tang was prepared.ICCs after stable culture were randomly divided into blank group,control group(normal rat serum),low concentration group(5%Banxia Xiexin Tang serum),medium concentration Group(10%Banxia Xiexin Tang serum)and high concentration group(20%Banxia Xiexin Tang serum).The drug serum was intervened at three time points of 12h、24h and 48h,respectively.Immunofluorescence double staining was used to observe the effect of Banxia Xiexin Tang on the phenotype maintenance of mouse ICCs by double-labeled three groups of marker proteins c-kit/α-SMA、c-kit/desmin and c-kit/SMMHC.The morphological expression differences,and explore the serum intervention conditions of Banxia Xiexin Tang.Western blotting was used to detect the protein expression levels of IP3R、RyR、PLC、c-kit、α-SMA、desmin、SMMHC and MLC-2 in ICCs.RT-PCR was used to detect IP3R mRNA,RyR3 mRNA and PLC mRNA in ICCs.Expression of c-kit mRNA、α-SMA mRNA、desmin mRNA、SMMHC mRNA and MLC-2 mRNA.[Results](1)ICC phenotype maintenance concentration:Compared with other groups,the positive expression of c-kit in ICCs in the high concentration group was significantly increased(p<0.05),while the positive expression of α-SMA was significantly decreased(p<0.05),SMMHC There was no significant difference in expression and desmin expression compared with normal blank(p>0.05).(2)Time of ICC phenotype maintenance:Compared with other groups,the positive expression of c-kit in ICCs was significantly increased in 12h group(p<0.05),and the expression of α-SMA,desmin and SMMHC was relatively small.Significance(p<0.05).(3)The expression level of related phenotypic proteins:1)Compared with the blank group,the protein expressions of α-SMA,SMMHC and MLC-2 in the drug group were significantly decreased(p<0.05),and the protein expression of c-kit in the drug group was significantly increased.High(p<0.05)),there was no significant change in the normal group d-SMA and SMMHC compared with the blank(p>0.05),and the difference was not statistically significant.2)Compared with the normal group,the expression of c-kit protein in the drug group was significantly increased(p<0.05),and the protein expression of α-SMA,SMMHC and MLC-2 in the drug group was significantly lower than that in the normal group(p<0.05).Statistically significant;3)There was no significant difference in protein expression between the three groups of Desmin(p>0.05),and the difference was not statistically.(4)The expression levels of related ion channel proteins:1)Compared with the blank group,the protein expressions of IP3R,RyR and PLC in the drug group were significantly increased(p<0.05).The normal group IP3R,PLC had no significant change compared with the blank.The difference was not statistically significant(p>0.05).The expression of RyR in the normal group was increased,the difference was statistically signifieant(p<0.05).2)Compared with the normal group,the protein expression of IP3R,RyR and PLC in the drug group was significantly higher than that in the normal group(p<0.05).Learning meaning.(5)Related phenotype protein mRNA expression:1)Compared with the blank group,the expression of c-kit mRNA was significantly increased in the normal group,and the expression of MLC-2 mRNA was significantly decreased(p<0.05).The difference was statistically significant.The expression of c-kit mRNA was significantly different from that of the blank group(p>0.05).The expressions of α-SMA,SMMHC and MLC-2 mRNA in the drug group were decreased(p<0.05).2)Compared with the normal group,the expressions of α-SMA mRNA,SMMHC mRNA and MLC-2 mRNA in the drug group were significantly lower than those in the normal group(p<0.05),and the expression of c-kit mRNA was significantly higher than that in the normal group(p<0.05).The difference was statistically significant;3)the expression of desmin mRNA was not statistically significant between the groups(p>0.05),and the difference was not statistically significant.(6)Expression of related ion channel protein mRNA:l)Compared with the blank group,there was no significant difference in the expression of IP3R mRNA,RyR3 mRNA and PLC mRNA in the normal group(p>0.05),and the expression of IP3R mRNA,RyR3 mRNA and PLC mRNA in the drug group.The difference between the blank group and the expression was significant(p<0.05),which was statistically significant.2)Compared with the normal group,the expression of IP3R mRNA,RyR3 mRNA and PLC mRNA in the drug group were significantly higher than those in the normal group(p<0.05).The difference is statistically significant.[Conclusion](1)Banxia Xiexin Tang could significantly promote the phenotype maintenance of ICC,especially in the high concentration group after 12 hours of drug intervention.(2)Banxia Xiexin Tang can significantly increase the protein and mRNA expression of c-kit,and decrease the expression of α-SMA,SMMHC,MLC-2 protein and mRNA,but there is no significant difference in the expression of desmin protein and mRNA.(3)Banxia Xiexin Tang can increase the expression of IP3R,RyR and PLC proteins in ICCs,and enhance the expression of IP3R mRNA,RyR3 mRNA and PLC mRNA related genes.In summary,Banxia Xiexin Tang can promote ICC phenotype maintenance,enhance c-kit protein expression,and promote calcium receptor protein and gene expression levels(IP3R,RyR3,PLC),affect related ion channels,and then coordinate The way to produce rhythmic contractions and spread through the GI muscle tissue,thereby achieving effective treatment to regulate gastrointestinal motility.On the molecular level,it provides a scientific basis for the clinical application of Banxia Xiexin Tang in the treatment of "heart sputum".