Study On The Intervention Of Banxia Xiexin Decoction On ICC Apoptosis Based On MiR-451-5p Targeting SCF/c-ki

[Objective]To explore the intervention effect of Banxia Xiexin Decoction on ICC apoptosis from the perspective of miR-451-5p targeted regulation of SCF/c-kit signaling pathway,so as to further clarify the cellular and molecular mechanism of Banxia Xiexin Decoction in the treatment of gastrointestinal motility disorders.[Methods](1)The serum of Banxia Xiexin Decoction was prepared.The ICC in logarithmic growth phase was randomly divided into blank group,normal group(containing 10%deionized water serum culture)and traditional Chinese medicine group(containing 10%Banxia Xiexin Decoction serum culture).The apoptosis of ICC in each group was detected by flow cytometry,and the expression of miR-451-5p was detected by qPCR.(2)ICC in logarithmic growth stage after passage was evenly inoculated on the culture plate for 24 hours to make it adhere to the wall.The cells were divided into blank group,Inhibitor group(transfected with miR-451-5p Inhibitor),Inhibitor NC group(transfected with miR-451-5p Inhibitor negative control),Mimics group(transfected with miR-451-5p Mimics),Mimics NC group(transfected with miR-451-5p Mimics negative control);Western Blot was used to detect the expression of SCF,c-kit,Bcl-2,Bax,Bad,PI3K,Akt and Caspase-9.(3)ICC in logarithmic growth phase were randomly divided into blank group,control group(transfected with miR-451-5p Inhibitor),traditional Chinese medicine group(containing 10%Banxia Xiexin Decoction serum culture)and normal group(containing 10%deionized water serum culture);The expressions of SCF,c-kit,Bcl-2,Bax,Bad,PI3K,Akt and Caspase-9 were detected by Western Blot;The mRNA expressions of SCF,c-kit,Bcl-2,Bax,PI3K and Akt were detected by RT-PCR.[Results](1)Compared with the blank group,the apoptosis rate of ICC in traditional Chinese medicine group was significantly decreased(P<0.05),and the expression of miR-451-5p was significantly increased(P<0.05).(2)Compared with the blank group,the apoptosis rate of Inhibitor group was significantly increased(P<0.05),the protein expressions of SCF,c-kit,Bcl-2,PI3K and Akt were significantly decreased(P<0.05),and the protein expressions of Bax,Bad and Caspase-9 were increased(P<0.05);In Mimics group,the apoptosis rate was significantly decreased(P<0.05),the protein expression of SCF,c-kit,Bax and Akt had no significant difference(P>0.05),the protein expression of Bcl-2 and PI3K was increased(P<0.05),and the protein expression of Bad and Caspase-9 was decreased(P<0.05).(3)Compared with the control group,the apoptosis rate of ICC in traditional Chinese medicine group was significantly decreased(P<0.05),the protein expressions of SCF,c-kit,Bcl-2,PI3K and Akt were significantly increased(P<0.05),and the protein expressions of Bax,Bad and Caspase-9 were significantly decreased(P<0.05).The mRNA expressions of SCF,c-kit,PI3K,Akt,Bcl-2 and Bax in traditional Chinese medicine group were significantly increased(P<0.05).[Conclusion]Banxia Xiexin Decoction can inhibit the expression of miR-451-5p,inhibit the apoptosis of ICC with inhibit the expression of miR-451-5p,enhance the expression of SCF/c-kit signaling pathway,PI3K/Akt signaling pathway and Bcl-2 protein,and inhibit the expression of Bax,Bad and Caspase-9 protein.miR-451-5p may regulate the cascade reaction between SCF/c-kit-PI3k/Akt-Bcl-2/Bax signaling pathway and intervene the apoptosis of ICC.