文献类型：期刊文献

中文题名：香砂六君子汤调控RhoA/ROCK2/MYPT1信号通路改善功能性消化不良大鼠胃动力的机制

英文题名：Mechanism of Xiangsha Liujunzi Tang in Regulation of Rho A/ROCK2/MYPT1 Pathway to Improve Gastric Motility in Rats with Functional Dyspepsia

作者：刘梦雅[1,2];成映霞[3];白敏[1,2];赵琳娜[1,2];李润法[1,2];安耀荣[1];段永强[3];李亚荣[3]

第一作者：刘梦雅

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃省实验动物行业技术中心,兰州730000;[3]宁夏医科大学中医学院,银川750004

第一机构：甘肃中医药大学

年份：2023

卷号：29

期号：4

起止页码：1

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：宁夏回族自治区科技厅重点研发引才专项(2022BSB03080);2022年甘肃中医药大学研究生创新创业基金资助项目(2022CX30)。

语种：中文

中文关键词：香砂六君子汤;RAS同源基因家族成员A(RhoA);Rho关联卷曲螺旋蛋白激酶2(ROCK2);磷酸化肌球蛋白轻链磷酸酶靶向亚基1(p-MYPT1);功能性消化不良

外文关键词：Xiangsha Liujunzi Tang;spleen and stomach deficiency type;Ras homolog gene family member A(RhoA);Rho associated coiled-coil containing protein kinase 2(ROCK2);phosphorylated-myosin phosphatase target subunit 1(p-MYPT1);functional dyspepsia

摘要：目的:基于RAS同源基因家族成员A(RhoA)/Rho关联卷曲螺旋蛋白激酶2(ROCK2)/肌球蛋白轻链磷酸酶靶向亚基1(MYPT1)信号通路探讨香砂六君子汤对功能性消化不良(FD)大鼠胃动力障碍的干预机制。方法:将60只SPF级雄性SD乳鼠随机分为正常组(n=10)和造模组(n=50),造模组大鼠采用综合造模法复制FD大鼠模型。模型复制成功后将造模组大鼠随机分为模型组、莫沙必利组和香砂六君子汤高、中、低剂量组,共5组,每组10只。正常组和模型组大鼠给予10 mL·kg^(-1)·d^(-1)生理盐水灌胃,莫沙必利组给予1.35 mg·kg^(-1)·d^(-1)莫沙必利灌胃,香砂六君子汤高、中、低剂量组分别给予12、6、3 g·kg^(-1)·d^(-1)香砂六君子汤汤剂灌胃,持续干预14 d。造模和给药前后观察大鼠一般生存状况,测定大鼠体质量、3 h平均进食量。给药干预结束后,测定大鼠肠推进率,采用苏木素-伊红(HE)染色观察胃组织病理改变,采用酶联免疫吸附测定法(ELISA)测定各组大鼠延髓和胃组织匀浆液中胆碱乙酰转移酶(ChAT)、血管活性肠肽(VIP)含量,采用冰冻切片染色技术观察各组大鼠胃窦平滑肌中腺嘌呤核苷三磷酸(ATP)酶分布,采用蛋白免疫印迹法检测胃组织中RhoA、ROCK2、磷酸化(p)-MYPT1蛋白表达水平。结果:与正常组比较,模型组大鼠毛发枯乱、倦懒怠动、行动缓迟,一般生存状况较差,体质量、3 h平均进食量以及肠推进率均明显下降(P<0.05);胃组织病理未见明显异常;延髓和胃组织中ChAT含量减少,胃组织中VIP含量增多,胃窦平滑肌中ATP酶分布明显减少,胃组织中RhoA、ROCK2、p-MYPT1蛋白表达水平均明显降低(P<0.05)。与模型组比较,各给药组大鼠一般生存状况明显好转,体质量、3 h平均进食量及肠推进率明显增加(P<0.05);胃组织病理未见明显差异;延髓和胃组织中ChAT含量明显增加,胃组织中VIP含量减少,胃窦平滑肌中ATP酶分布明显增加,胃组织中RhoA、ROCK2、p-MYPT1蛋白表达水平明显升高(P<0.05),上述指标香砂六君子汤组干预作用呈剂量依赖性。结论:香砂六君子汤能够有效改善FD大鼠一般生存状况及胃动力,其具体机制可能与香砂六君子汤激活胃组织中RhoA/ROCK2/MYPT1信号通路调控平滑肌舒缩,促进胃动力有关。
Objective:To explore the intervention mechanism of Xiangsha Liujunzi Tang in rats with functional dyspepsia(FD)based on the Ras homolog gene family member A(RhoA)/Rho-associated coiled-coil containing protein kinase 2(ROCK2)/Myosin phosphatase target Subunit 1(MYPT1)pathway.Method:Sixty male SD suckling rats in SPF grades were randomly divided into blank group(n=10)and model group(n=50).The comprehensive modeling method(gavage administration of iodoacetamide+exhaustion of swimming+disturbance of hunger and satiety)was used to replicate the rat model of FD.After successful replication of the model,the rats in the model group were randomly divided into model group,mosapride group,and high,middle,and low-dose Xiangsha Liujunzi Tang groups,with 10 rats in each group.Rats in the blank group and model group were given 10 mL kg^(-1)·d^(-1)normal saline,those in the mosapride group were given 1.35 mg·kg^(-1)·d^(-1)mosapride,and those in the high,middle,and low-dose Xiangsha Liujunzi Tang groups were given 12,6,and 3 g·kg^(-1)·d^(-1)Xiangsha Liujunzi Tang,respectively.The intervention lasted 14 days.The general living conditions of rats were observed before and after modeling and administration,and the 3-hour food intake and body mass of rats were measured.After intervention,the intestinal propulsion rate of rats was measured,and the pathological changes in the gastric tissue were observed by hematoxylin-eosin(HE)staining.The content of choline acetyl transferase(ChAT)and vasoactive intestinal peptide(VIP)in the medulla oblongata and gastric tissue homogenate was determined by enzyme-linked immunosorbent assay(ELISA),the distribution of adenosine triphosphate(ATP)enzyme in gastric antrum smooth muscle was observed by frozen section staining,and the protein expression levels of RhoA,ROCK2,and phosphorylated-myosin phosphatase target subunit 1(pMYPT1)in the gastric tissue were detected by Western blot.Result:Compared with the blank group,the model group had withered hair,lazy movement,slow action,poor general living condition,lower 3-hour food intake,body mass,and lower intestinal propulsion rate(P<0.05),whereas no obvious abnormality in gastric histopathology.In the model group,the content of ChAT in the medulla oblongata and gastric tissue decreased,the content of VIP in gastric tissue increased,the distribution of ATP enzyme in gastric antrum smooth muscle decreased significantly,and the protein expression levels of RhoA,ROCK2,and p-MYPT1 in the gastric tissue decreased significantly(P<0.05).As compared with the model group,the general living condition of rats in each intervention group was significantly improved,and the 3-hour food intake,body mass,and intestinal propulsion rate were significantly increased(P<0.05).There was no significant difference in gastric pathology in the intervention groups.The content of ChAT in the medulla oblongata and gastric tissue increased significantly,the content of VIP in the gastric tissue decreased,the distribution of ATP enzyme in gastric antrum smooth muscle increased significantly,and the protein expression levels of RhoA,ROCK2,and p-MYPT1 in the gastric tissue increased significantly(P<0.05).The intervention effect of Xiangsha Liujunzi Tang group on the above indexes was dose-dependent.Conclusion:Xiangsha Liujunzi Tang can effectively improve the general living condition and gastric motility of rats with FD,and its specific mechanism may be related to the activation of the RhoA/ROCK2/MYPT1 pathway in the gastric tissue to regulate smooth muscle relaxation and contraction and promote gastric motility.