Effect Of Shenmai Injection On Expression Of TRPA1 And Proliferation In Airway Smooth Muscle Cells Of Asthmatic Rats

ObjectiveTo investigate the effect of Shenmai injection on the expression of transient receptor potential channel protein A1(TRPA1)in airway smooth muscle cells(ASMCs)of asthmatic rats and its possible mechanism of proliferation regulation of ASMCs in asthmatic rats.MethodsA total of 30 six-week-old male SPF SD rats were randomly divided into three groups according to the random number table: control group,asthma group and Shenmai injection treatment group,with 10 rats in each group.In the asthma group and the Shenmai injection group,the asthma model was established by using ovalbumin.In the Shenmai injection group,2 ml of Shenmai injection was injected into the peritoneal cavity every day,and the lung tissue and tail vein blood were taken after the last stimulation operation.The changes of airway structure in each group were detected by HE staining,and the wall area(WA)/intraluminal circumference(Pi),bronchial smooth muscle area(S)/Pi,bronchial smooth muscle cell number(N)/Pi were calculated.ASMCs were identified by immunocytochemistry and immunofluorescence;TRPA1 and PCNA m RNA expression was detected by RT-PCR;TRPA1 protein expression was detected by Western blotting;PCNA protein expression was detected by immunocytochemistry;the concentration of Ca2+ in ASMCs was detected by laser confocal microscopy;the proliferation of ASMCs in each group was detected by CCK-8 method;the levels of IL-4,IL-5 and IL-13 in tail vein blood were detected by ELISA.The SPSS 22.0 was used for all statistical analysis,one-way ANOVA was used for comparison among multiple groups,and LSD-t method was used for pairwise comparison among groups.ResultsCompared with the control group,the airway structure of the asthma group was remodeled,and the WA/Pi,S/Pi,and N/Pi of the trachea were significantly increased(P<0.05).Compared with the asthma group,airway remodeling was inhibited in the Shenmai injection group,and the WA/Pi,S/Pi,and N/Pi of the trachea were significantly decreased(P<0.05).Compared with the control group,the expression of TRPA1 and PCNA m RNA and protein in ASMCs of asthmatic rats was increased,intracellular Ca2+ concentration was increased,proliferation was increased,and levels of IL-4,IL-5 and IL-13 in tail vein blood were increased(all P<0.05).Compared with the asthma group,the expression of TRPA1 and PCNA m RNA and protein in ASMCs of Shenmai injection treatment group was significantly reduced,intracellular Ca2+ concentration was decreased,proliferation was decreased,and levels of IL-4,IL-5 and IL-13 in tail vein blood were decreased(all P<0.05).ConclusionsShenmai injection may prevent the excessive proliferation of ASMCs in asthma by inhibiting the expression of TRPA1 channel which regulates intracellular Ca2+ concentration and levels of IL-4,IL-5,IL-13.