逍遥散对肝郁脾虚证大鼠海马组织RhoA/ROCK2通路的影响  

Effect of Xiaoyao Powder on RhoA/ROCK2 Pathway in Hippocampus of Rats with Liver Depression and Spleen Deficiency Syndrome

作　　者：郑于林 陈嫒 李熠毅[1] 高琳[1] 李晓红[1,2] 杨力强[1,2] ZHENG Yulin;CHEN Ai;LI Yiyi;GAO Lin;LI Xiaohong;YANG Liqiang(Guangxi University of Chinese Medicine,Nanning 530200,China;Guangxi Key Laboratory of Chinese Medicine Foundation Research,Guangxi University of Chinese Medicine,Nanning 530200,China)

机构地区：[1]广西中医药大学,南宁530200 [2]广西中医药大学广西中医基础研究重点实验室,南宁530200

出　　处：《中国中医基础医学杂志》2023年第6期929-932,共4页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金项目(81860817);国家自然科学基金项目(81560750);广西中医药大学广西中医基础研究重点实验室自主研究课题(20-065-53-06);广西中医药大学研究生教育创新计划项目(YCXJ2021002)。

摘　　要：目的 探究肝郁脾虚证的生物学基础及逍遥散的作用机制。方法 采用慢性束缚应激方法建立肝郁脾虚证大鼠模型。72只SD雄性大鼠随机分为6组,正常组,模型组,氟西汀组(0.0018 g/kg),逍遥散高(16.7 g/kg)、中(8.35 g/kg)、低(4.175g/kg)剂量组,每组12只。造模结束后,ELISA法测定大鼠血清单丝氨酸蛋白激酶1(single serine protein kinase 1,LIMK1)的含量,荧光定量PCR、 Western blot检测海马组织RAS同源基因家族成员A(RAS homologous gene family member A,RhoA)、Rho相关螺旋卷曲蛋白激酶2(Rho-related spiral coiled protein kinase 2,ROCK2)的m RNA及蛋白表达。结果 模型组大鼠血清LIMK1含量,海马组织中RhoA、ROCK2的mRNA和蛋白表达水平均明显高于正常组(P<0.01);各药物组大鼠血清LIMK1含量明显低于模型组(P<0.01),逍遥散高、中剂量组和氟西汀组大鼠海马组织中RhoA、ROCK2的mRNA和蛋白表达明显低于模型组(P<0.05,P<0.01)。结论 慢性束缚应激肝郁脾虚证大鼠海马组织RhoA/ROCK2通路被激活,逍遥散可抑制RhoA/ROCK2通路激活,并且作用呈剂量依赖性。Objective To explore the biological basis of liver depression and spleen deficiency syndrome and the mechanism of Xiaoyao Powder.Methods The rat model of liver stagnation and spleen deficiency syndrome was established by chronic restraint stress.72 male SD rats were randomly divided into 6 groups:normal group,model group,fluoxetine group(0.0018g·kg-1),Xiaoyao Powder high(16.7g·kg-1),medium(8.35g·kg-1)and low(4.175g·kg-1)dose group,with 12 rats in each group.After modeling,the content of single serine protein kinase 1(LIMK1)in rat serum was measured by ELISA,and the gene and protein expressions of Ras homologous gene family member A(RhoA)and Rho related spiral coiled protein kinase 2(Rock2)in hippocampus were detected by real-time qPCR and Western blot.Results The serum LIMK1 content in the model group was significantly increased,and the gene and protein expressions of RhoA and ROCK2 in the hippocampus were significantly increased in comparison with the normal group(P<0.01).The serum LIMK1 content of each drug group was significantly lower,and the mRNA and protein expressions of RhoA and ROCK2 in hippocampus of high and medium dose Xiaoyao Powder groups and fluoxetine group were significantly lower in comparison with the model group(P<0.05,P<0.01).Conclusion RhoA/ROCK2 pathway was activated in hippocampus of rats with liver stagnation and spleen deficiency syndrome under chronic restraint stress.Xiaoyao Powder can inhibit RhoA/ROCK2 pathway activation in a dose-dependent manner.

关 键 词：肝郁脾虚证 慢性束缚应激 LIMK1 RhoA/ROCK2通路 逍遥散 

分 类 号：R289.5[医药卫生—方剂学]