论文部分内容阅读
目的 探究疏肝理气法对胃Cajal间质细胞凋亡的调控作用及可能的作用机制.方法 选用120只Wist-ar大鼠,平均分为三组进行处理:对照组,阳明腑实证组和疏肝理气法治疗组.阳明腑实证造模成功后,疏肝理气法治疗组每日灌服柴胡疏肝散水煎剂,连续灌胃处理5 d,阳明腑实证组和对照组每日灌服等量的0.9％氯化钠,连续处理5 d.处理20 d后,观察各组大鼠的存活率,检测存活大鼠的平均体重;利用放射免疫法检测血浆P物质(SP)水平;分离各组大鼠胃Cajal间质细胞,利用流式细胞仪检测各组细胞凋亡情况;Western-blot检测胃Cajal间质细胞中凋亡相关蛋白和PI3K/PDK1/Akt信号通路相关蛋白表达情况.结果 处理20 d后,阳明腑实证组大鼠存活率、存活大鼠平均体重、SP含量均显著低于对照组(P<0.05),疏肝理气法治疗组大鼠存活率、存活大鼠平均体重、SP含量显著高于阳明腑实证组(P<0.05).阳明腑实证组细胞凋亡率及Bax、Cleaved-caspase3、Cleaved-caspase9蛋白表达显著高于对照组,Bcl-2、PI3K、PDK1、Akt、p-Akt蛋白表达显著低于对照组(P<0.05),疏肝理气法治疗组细胞凋亡率及Bax、Cleaved-caspase3、Cleaved-caspase9蛋白表达均显著低于阳明腑实证组,Bcl-2、PI3K、PDK1、Akt、p-Akt蛋白表达显著高于阳明腑实证组(P<0.05).结论 疏肝理气法可能通过PI3K/PDK1/Akt信号通路减少由阳明腑实证引起的胃Cajal间质细胞凋亡,有效改善胃肠功能.“,”Objective To investigate the effect of soothing liver and regulating the qi method on gastric Cajal interstitial cell apoptosis and its possible mechanism. Methods 120 Wistar rats were divided into three groups:control group,yangmingfushi syndrome group and soothing liv-er and regulating the qi method group. After the success of the model,the patients in the treatment group were treated with Chaihu Shugan San De-coction,and the rats in the control group and the control group were fed with the same amount of 0. 9％ Sodium chloride,continuous treatment for 5 d. After 20 days treatment,the survival rate of rats in each group was observed,and the average weight of the surviving rats was measured. Plas-ma substance P (SP)levels were measured by radioimmunoassay. The Cajal interstitial cells in the stomach of each group rats were separated,and the apoptosis of each group was detected by flow cytometry. The expression of apoptosis related proteins and PI3K/ PDK1 / Akt signaling pathway re-lated proteins in stomach Cajal cells of was detected by Western-blot. Results After 20 days,the survival rate,the average body weight and SP content of the rats in the yangmingfushi group were significantly lower than those in the control group (P < 0. 05). The survival rate,the average body weight and SP content of the rats treated with soothing liver and regulating qi method were significantly higher than those of the yangmingfushi Group (P < 0. 05). The apoptosis rate and Bax,Cleaved-caspase3,Cleaved-caspase9 protein expression in the soothing liver and regulating the qi method group were significantly higher than those in the control group,and the expression of Bcl-2,PI3K,PDK1,Akt and p-Akt protein was significantly lower than that in the control group (P < 0. 05);cell apoptosis rate and Bax,Cleaved-caspase3 and Cleaved-caspase9 protein expression in soothing liver and regulating the qi method group were significantly lower than that of yangmingfushi syndrome group,the expression of Bcl-2,PI3K,PDK1,Akt,p-Akt protein was significantly higher than yangmingfushi syndrome group (P < 0. 05). Conclusion Soothing liver and regulating the qi method may reduce the apoptosis of Cajal interstitial cells which induced by the yangmingfushi syndrome through the PI3K/ PDK1 / Akt signal pathway,and effectively improve the gastrointestinal function.