摘要
目的:以疏肝健脾为核心,运用逍遥散加减治疗高泌乳素血症(HPRL)模型大鼠,探讨逍遥散加减对HPRL大鼠模型中枢多巴胺受体细胞外调节蛋白激酶(ERK)信号转导通路的作用机制。方法:96只SD雌性大鼠,随机分为正常组,模型组,溴隐亭组(0. 001 g·kg-1),逍遥散加减高、中、低剂量组(60,30,15 g·kg-1),多巴胺第一受体(D1R)拮抗剂组(SCH23390组,SCH23390+逍遥散加减中剂量组,SCH23390+溴隐亭组),多巴胺第二受体(D2R)拮抗剂组(氟哌啶醇组,氟哌啶醇组+逍遥散加减中剂量组,氟哌啶醇组+溴隐亭组)。采用垂体移植法制作HPRL大鼠模型,给药30 d后,采用蛋白免疫印迹法(Western blot)检测大鼠下丘脑癌蛋白Ras,Raf蛋白的表达;采用实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠下丘脑丝裂原活化蛋白激酶(MAPK)激酶1/2(MEK1/2) mRNA,细胞外信号调节激酶1/2(ERK1/2) mRNA的表达;采用电镜观察卵巢颗粒细胞超微结构。结果:与正常组比较,模型组Ras,Raf蛋白,MEK1/2,ERK1/2 mRNA表达显著降低(P <0. 01);运用SCH23390拮抗DIR后,与模型组比较,SCH23390组Ras,Raf蛋白,MEK1/2,ERK1/2 mRNA表达有升高趋势,运用氟哌啶醇拮抗D2R后,与模型组比较,氟哌啶醇组Ras,Raf蛋白表达有下降趋势,MEK1/2,ERK1/2 mRNA表达显著下降(P <0. 01);运用逍遥散加减治疗后,能显著提高Ras,Raf蛋白,MEK1/2,ERK1/2 mRNA表达(P <0. 01),且逍遥散加减高、中剂量组表达增加最明显(P <0. 01),效果优于溴隐亭组。电镜下模型组卵巢颗粒细胞形态及线粒体数量、结构异常,运用逍遥散加减治疗后,颗粒细胞形态和线粒体结构趋于正常。结论:逍遥散加减通过激动D2R,抑制D1R,调节ERK/MAPK信号通路,并有效改善卵巢颗粒细胞线粒体功能,促进卵泡发育,说明逍遥散加减治疗HPRL是多靶点的作用机制。
Objective:To explore the mechanism of modified Xiaoyaosan on extracellular regulated protein kinase(ERK)signal transduction pathway of central dopamine receptor in hyperprolactinemia(HPRL)model rats by treating them with Shugan Jianpi as the core.Method:The 96 SD female rats were randomly divided into normal group,model group,Bromocriptine group(0.001 g·kg-1),modified Xiaoyaosan high,medium and low dose group(60,30,15 g·kg-1),dopamine D1 receptor(D1 R)antagonist group(SCH23390,SCH23390+modified Xiaoyaosan group,SCH23390+bromocriptine group),dopamine D2 receptor(D2R)antagonist group(haloperidol group,haloperidol+modified Xiaoyaosan group,haloperidol+bromocriptine group).HPRL rat model was established by pituitary transplanta-tion.After 30 days of administration,renin-angiotensinsystem Ras and Raf protein expressions in thypothalamus of rats were detected by Western blot,and mitogen-activated protein kinase(MAPK)kinase 1/2(MEK1/2)mRNA and extracellular signaling regulates kinase 1/2(ERK1/2)mRNA expressions in the hypothalamus of rats were detected by Real-time PCR.The microstructures of mitochondria in ovarian granulosa cells were observed by electron microscopy.Result:The Ras,Raf protein,MEK1/2 and ERK1/2 mRNA expression were significantly decreased in model group compared with normal group(P<0.01).After SCH23390 antagonistic DIR was applied,Ras,Raf pprotein,MEK1/2 and ERK1/2 mRNA expression in SCH23390 group showed an increased trend compared with model group,and when haloperidol was used to antagonistic D2R,Ras and Raf expression in haloperidol group showed a decreased trend compared with model group,and MEK1/2 and ERK1/2 mRNA expression significantly decreased(P<0.01).The expression of Ras,Raf protein,MEK1/2 and ERK1/2 mRNA was significantly increased after using modified Xiaoyaosan(P<0.01),and the expression was significantly increased in the high and medium-dose groups of prescription(P<0.01).The effect was better than that of bromocriptine group.The morphology of granulosa cells and the number and structur
作者
许露
李燕
张芳
陈婧
XU Lu;LI Yan;ZHANG Fang;CHEN Jing(Traditional Chinese Hospital of Liuan,Liuan 237000,China;The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2019年第15期70-76,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81260549)
