摘要
目的探讨逍遥散加减方治疗肝郁脾虚型高催乳素血症(HPRL)的可能作用机制。方法96只SD雌性大鼠随机分为正常组16只,造模组80只。造模组大鼠采用慢性不可预知应激联合甲氧氯普胺腹腔注射构建HPRL肝郁脾虚型模型大鼠。将80只造模成功的大鼠随机分为模型组,逍遥散加减方高、中、低剂量组和甲磺酸溴隐亭组,每组16只。逍遥散加减方高、中、低剂量组分别给予逍遥散加减方60、30、15 g/(kg·d)灌胃,甲磺酸溴隐亭组予以甲磺酸溴隐亭片1 mg/(kg·d)灌胃,正常组及模型组给予生理盐水10 ml/(kg·d)灌胃,连续给药14天。ELISA法检测血清催乳素(PRL)含量;免疫组化测定大鼠下丘脑肿瘤坏死因子α(TNF-α)和酪氨酸羟化酶(TH)表达;采用Western blot法检测下丘脑肿瘤坏死因子受体1(TNFR1)蛋白表达;RT-PCR法检测下丘脑受体相互作用蛋白激酶3(RIP3)mRNA表达;免疫荧光法检测RIP3和多巴胺能神经元的共表达情况。结果与正常组比较,模型组血清PRL含量升高,下丘脑TNF-α、TNFR1、RIP3mRNA及RIP3与多巴胺能神经元共表达阳性细胞数显著升高,TH表达降低(P<0.05或P<0.01);与模型组比较,甲磺酸溴隐亭组和逍遥散加减方低剂量组下丘脑TNF-α表达降低,甲磺酸溴隐亭组和逍遥散加减方中、高剂量组TH表达显著升高,逍遥散加减方各剂量组和甲磺酸溴隐亭组血清PRL含量,下丘脑TNFR1、RIP3 mRNA,RIP3与多巴胺能神经元共表达阳性细胞数均显著降低(P<0.05或P<0.01);与甲磺酸溴隐亭组比较,逍遥散加减方高、低剂量组血清PRL含量显著升高,逍遥散加减方中剂量组TH表达显著升高,逍遥散加减方低剂量组下丘脑RIP3 mRNA表达降低,逍遥散加减方高剂量组RIP3与多巴胺能神经元共表达阳性细胞数显著升高(P<0.01)。结论逍遥散加减方可调节下丘脑多巴胺能神经元细胞程序性坏死,影响DA表达,从而调节PRL水平,可能是其治疗HPRL肝郁脾虚型�
Objective To investigate the possible mechanisms of modified Xiaoyao Powder(逍遥散)in the treatment of hyperprolactinemia(HPRL)with liver constraint and spleen deficiency.Methods Ninety-six female SD rats were randomly divided into a normal group(n=16)and a modeling group(n=80).In the modeling group,rats were subjected to chronic unpredictable stress combined with intraperitoneal injection of metoclopramide to establish a rat model of HPRL with liver constraint and spleen deficiency.The 80 successfully modeled rats were randomly divided into a model group,a high,medium,and low-dose group of modified Xiaoyao Powder,and a bromocriptine group,with 16 rats in each group.The high,medium,and low-dose groups of modified Xiaoyao Powder were orally administered doses of 60,30,and 15 g(/kg·d)respectively,the bromocriptine group was orally administered bromocriptine tablets at a dose of 1 mg/(kg·d),and the normal group and model group were orally administered 10 ml/(kg·d)of normal saline for 14 consecutive days.ELISA was used to detect serum prolactin(PRL)level;immunohistochemistry was used to determine the expression of tumor necrosis factor-alpha(TNF-α)and tyrosine hydroxylase(TH)in the hypo⁃thalamus;Western blot was used to detect the protein expression of tumor necrosis factor receptor 1(TNFR1)in the hypothalamus;Real-time PCR was used to detect the mRNA expression of receptor interacting protein kinase 3(RIP3)in the hypothalamus;immunofluorescence was used to detect the co-expression of RIP3 and dopamine neurons in the hypothalamus.Results Compared with the normal group,the serum PRL levels were increased in the model group,and the expression of hypothalamic TNF-α,TNFR1,RIP3 mRNA,and the co-expression of RIP3 with dopamine neurons were significantly increased,while TH expression was decreased(P<0.05 or P<0.01).Compared with the model group,the expression of hypothalamic TNF-αwas decreased in the bromocriptine group and low-dose group of modified Xiaoyao Powder,and the expression of TH was significantly incre
作者
李燕
李燕
刘凯鑫
代康莉
刘晓
杨惠
罗丹
LI Yan;LI Yan;LIU Kaixin;DAI Kangli;LIU Xiao;Yang Hui;Luo Dan(Guizhou University of Traditional Chinese Medicine,Guiyang,550001)
出处
《中医杂志》
CSCD
北大核心
2024年第3期317-323,共7页
Journal of Traditional Chinese Medicine
基金
国家自然科学基金(81774361)。
关键词
高催乳素血症
逍遥散
下丘脑
多巴胺能神经元
疏肝健脾法
程序性坏死
hyperprolactinemia
Xiaoyao Powder
hypothalamus
dopaminergic neurons
soothe the liver and fortify the spleen
programmed cell death
