Huang Qing-Fang, Gong Meng-Juan, Chen Yan-Fen, Feng Cheng-En
School of Chemistry and Chemical Engineering, Guangdong Pharmaceutical University Zhongshan 528458, China.
School of Traditional Chinese Medicine, Guangdong Pharmaceutical University Guangzhou 510006, China.
Zhongguo Zhong Yao Za Zhi. 2020 May;45(9):2144-2150. doi: 10.19540/j.cnki.cjcmm.20200213.401.
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 Ca(2+)-Mg(2+)-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, Ca(2+)-Mg(2+)-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.
本文旨在探讨藿香正气口服液对湿阻脾胃证大鼠肠道屏障功能的影响，并初步探究其作用机制。建立湿阻脾胃证大鼠模型，然后将造模成功的大鼠按性别和体重随机分为模型对照组、藿香正气口服液高、低剂量组和自然恢复组，每组10只。另取10只大鼠作为空白对照组。各组给予相应处理7天后，分离大鼠血清。采用MTT法检测D-乳酸含量，采用速率法检测二胺氧化酶（DAO）活性。分离大鼠结肠组织，采用磷酸测定法检测Na⁺-K⁺-ATP酶活性和Ca²⁺-Mg²⁺-ATP酶活性，采用分光光度法检测谷胱甘肽过氧化物酶（GSH-Px）活性，采用钼酸铵法检测过氧化氢酶（CAT）活性，采用羟胺法检测超氧化物歧化酶（SOD）活性，采用免疫荧光法检测闭合蛋白（occludin）和紧密连接蛋白1（ZO-1）的表达，采用蛋白质印迹法检测occludin蛋白和ZO-1蛋白的表达水平。结果显示：低剂量藿香正气口服液能明显改善湿阻脾胃证大鼠的体重、饮食、大便及小便状态。湿阻脾胃证大鼠血清中D-乳酸含量及DAO活性明显降低。大鼠结肠组织中Na⁺-K⁺-ATP酶、Ca²⁺-Mg²⁺-ATP酶、GSH-Px、CAT及SOD活性明显升高。大鼠结肠组织中occludin蛋白和ZO-1蛋白表达水平明显升高。藿香正气口服液组与模型对照组上述指标比较，差异有统计学意义（P＜0.05）。藿香正气口服液能有效恢复湿阻脾胃证大鼠的肠道屏障功能，其机制可能与肠道机械屏障功能的修复有关。
