舒胃汤对FD肝郁脾虚证模型大鼠胃肠平滑肌Beclin-1、mTOR、Cx43蛋白与mRNA表达的影响

Effects of Shuwei Decoction on the expressions of Beclin-1,mTOR,Cx43 Protein and mRNA in gastrointestinal smooth muscle of rats with functional dyspepsia due to liver depression and spleen deficiency

目的通过研究舒胃汤(柴胡、香附、白术,等)对功能性消化不良(FD)肝郁脾虚证模型大鼠胃窦和幽门区平滑肌中自噬调控基因Beclin-1、雷帕霉素靶蛋白mTOR、连接蛋白Cx43蛋白与mRNA表达的影响,从自噬与细胞间缝隙连接通道探讨舒胃汤对功能性消化不良的治疗作用及机制。方法取60只Wistar大鼠随机分为空白组,模型组,莫沙必利组(1.37 mg/kg),舒胃汤低、中、高剂量组(7.67、15.34、30.68 g/kg)。除空白组外,各组大鼠按复合病因造模法造模21 d复制FD肝郁脾虚证大鼠模型。于造模结束后第1天开始给药,空白组与模型组大鼠给予等量蒸馏水,其余各组大鼠按剂量要求灌胃给药,每天1次,连续给药14 d。末次给药2 h后,颈静脉放血处死大鼠,摘取各组大鼠胃窦与幽门组织,Western blot法检测Beclin-1、mTOR、Cx43蛋白表达,RT-qPCR法检测Beclin-1、mTOR、Cx43 mRNA表达。结果 Western blot结果显示,舒胃汤低、中、高剂量组Beclin-1表达明显降低(P<0.01),舒胃汤高剂量组Cx43表达明显升高(P<0.05);RT-qPCR结果显示,舒胃汤中、高剂量组Beclin-1表达明显降低(P<0.01),mTOR表达明显升高(P<0.01),舒胃汤低、中、高剂量组Cx43表达明显升高(P<0.01)。结论舒胃汤可通过上调FD模型大鼠胃肠平滑肌细胞mTOR mRNA、Cx43蛋白及mRNA表达,下调Beclin-1蛋白及mRNA表达,抑制细胞自噬及增强细胞间缝隙连接治疗功能性消化不良。

AIM To investigate the therapeutic action and mechanism of Shuwei Decoction（ Bupleuri Radix,Cyperi Rhizoma,Atractylodis macrocephalae Rhizoma,etc.） on functional dyspepsia（ FD） from the perspectives of autophagy and intercellular gap junction pathway through its impact on the expressions of Beclin-1,mTOR,Cx43 protein and mRNA in gastric antrum pyloric smooth muscle cells of FD model rats. METHODS Sixty Wistar rats were randomly divided into control（ blank） group,model group,mosapride group（ 1. 37 mg/kg）,low,medium and high dose Shuwei Decoction groups（ 7. 67 g/kg,15. 34 g/kg,30. 68 g/kg,respectively）. FD model rats were prepared according to compound etiological modeling method for 21 days. On the first day after modeling,the rats in each group were given the corresponding medicine for intragastric administration once a day for 14 days. Two hours after the last administration,we procured gastric antrum and pyloric tissues from the sacrificed rats. The expressions of Beclin-1,mTOR and Cx43 protein were detected by Western blot,and so were the expressions of Beclin-1,mTOR and Cx43 mRNA by quantitative real-time PCR（ RT-qPCR）. RESULTS We observed decreased expression of Beclin-1 protein in low,medium and high dose Shuwei Decoction groups（ P〈0. 01）,and an increased expression of Cx43 protein in high dose Shuwei Decoction group（ P〈0. 05）. In medium and high dose Shuwei Decoction groups,decreased expression of Beclin-1 mRNA（ P〈0. 01）,and increased expression of mTOR and Cx43 mRNA（ P〈0. 01） were detected as well. CONCLUSION Shuwei Decoction can improve FD by suppressing cell autophagy and enhancing the function of gap junction through up-regulating the expression of mTOR mRNA and Cx43 protein and mRNA,and down-regulating the expression of Beclin-1 protein and mRNA.