藿香正气口服液对湿困脾胃证大鼠肠屏障功能的作用研究

研究藿香正气口服液对湿困脾胃证大鼠肠屏障功能的作用并初步探讨作用机制。建立湿困脾胃证大鼠模型,将成模大鼠按性别、体质量随机分为模型组,藿香正气口服液高、低剂量组,自然恢复组,每组10只,另取10只为空白组。每组给予相应治疗7 d后,分离大鼠血清,MTT法检测D-乳酸含量,速率法检测二胺氧化酶(DAO)活性;分离大鼠结肠组织,定磷法检测Na^+-K^+-ATP酶活性和Ca²⁺-Mg²⁺-ATP酶活性,比色法检测谷胱甘肽过氧化物酶(GSH-Px)活性,钼酸铵法检测过氧化氢酶(CAT)活性,羟胺法检测超氧化物歧化酶(SOD)活性,免疫荧光法检测occludin和紧密连接蛋白ZO-1的表达,Western blot法检测occludin蛋白和ZO-1蛋白的表达水平。结果显示,藿香正气口服液低剂量组能明显改善湿困脾胃证大鼠体质量、饮食、二便等状态,降低湿困脾胃证大鼠血清中D-乳酸含量和DAO活性,升高湿困脾胃证大鼠结肠组织Na^+-K^+-ATP酶活性、Ca²⁺-Mg²⁺-ATP酶活性、GSH-Px活性、CAT活性和SOD活性,升高湿困脾胃证大鼠结肠组织occludin蛋白和ZO-1蛋白的表达水平,上述指标与模型组相比差异有统计学意义(P<0.05)。藿香正气口服液能有效恢复湿困脾胃证大鼠肠屏障功能,可能通过修复肠机械屏障功能发挥作用。

Effects of Huoxiang Zhengqi Oral Liquid on intestinal barrier function in rats with dampness obstructing spleen-stomach syndrome

The aim of this paper was to investigate the effect of Huoxiang Zhengqi Oral Liquid on intestinal barrier functions in rats with dampness obstructing spleen-stomach syndrome and primarily explore the mechanism.The rat model of dampness obstructing spleen-stomach syndrome was established,and then the modeled rats were randomly divided into the model control group,Huoxiang Zhengqi Oral Liquid high and low dose groups,and natural recovery group according to gender and body weight,with 10 rats in each group.Another 10 rats were taken as blank control group.After each group received the corresponding treatment for 7 days,rat serum was isolated.D-lactic acid content was detected by the MTT method,and diamine oxidase(DAO)activity was detected by the rate method.Colon tissues of the rats were isolated to detect Na+-K+-ATPase activity and Ca2+-Mg2+-ATPase activity by phosphate determination method,glutathione peroxidase(GSH-Px)activity was detected by spectrophotometry,catalase(CAT)activity was detected by ammonium molybdate,superoxide dismutase(SOD)activity was detected by hydroxylamine,the expression of occludin protein and ZO-1 protein was detected by immunofluorescence,and the expression levels of occludin protein and ZO-1 protein were detected by Western blot.Results showed that low dose Huoxiang Zhengqi Oral Liquid could improve the body weight,diet,stool and urine state of rats with dampness obstructing spleen-stomach syndrome obviously.The D-lactic acid content and the DAO activity in the serum of rats with dampness obstructing spleen-stomach syndrome were reduced obviously.The activities of Na+-K+-ATPase,Ca2+-Mg2+-ATPase,GSH-Px,CAT and SOD in rat colon tissues were increased obviously.The occludin proteins and ZO-1 protein expression levels in rat colon tissues were raised obviously.The differences in the above indexes between Huoxiang Zhengqi Oral Liquid group and the model control group were statistically significant(P<0.05).Huoxiang Zhengqi Oral Liquid could effectively restore the intestinal barrier function in rats with dampness obstructing spleen-stomach syndrome and its mechanism may be related to the repair of intestinal mechanical barrier function.