基于cAMP/PKA/CREB信号通路探讨柴胡皂苷对多发性抽动症小鼠的治疗作用  

Therapeutic effect of saikosaponin on Tourette syndrome mice based on cAMP/PKA/CREB signaling pathway

作　　者：陈羽 王佳斌[2] 颜建宏 张颖 CHEN Yu;WANG Jiabin;YAN Jianhong;ZHANG Ying(The First Department of Pediatrics,Hebei Cangzhou Integrated Traditional Chinese and Western Medicine Hospital,Cangzhou 061000,China;不详)

机构地区：[1]河北省沧州中西医结合医院儿一科,河北沧州061000 [2]河北省沧州中西医结合医院影像科 [3]任丘市人民医院儿科

出　　处：《山东医药》2024年第19期25-29,共5页Shandong Medical Journal

基　　金：河北省中医药类科学研究课题计划项目(2023251)。

摘　　要：目的基于环磷酸腺苷/蛋白激酶A/环磷酸腺苷反应成分结合蛋白(cAMP/PKA/CREB)信号通路探讨柴胡皂苷(SS)对多发性抽动症(TS)模型小鼠的治疗作用。方法将小鼠随机分为对照组、TS组、SS低剂量(SS-L)组、SS高剂量(SS-H)组、阳性药物组、SS-H+PKA抑制剂(H-89)组,每组10只。除对照组外,其他各组经腹腔注射亚氨基二丙腈溶液建立TS小鼠模型。造模后,SS-L组、SS-H组分别给予25、100 mg/kg SS腹腔注射,并以生理盐水灌胃;阳性药物组以0.5 mg/kg氟哌啶醇灌胃,并以生理盐水腹腔注射;SS-H+H-89组以100 mg/kg SS、5 mg/kg H-89腹腔注射,并以生理盐水灌胃;TS组及对照组分别以等体积的生理盐水腹腔注射、灌胃。每天1次,连续干预3周。对小鼠运动行为、刻板行为进行评分,用ELISA法检测纹状体组织中5-羟色胺(5-HT)、去甲肾上腺素(NE)以及多巴胺(DA),用苏木精-伊红法观察脑组织形态学变化,用免疫组化法检测黑质中酪氨酸羟化酶(TH)阳性神经元,用Western blotting法检测cAMP/PKA/CREB通路相关蛋白表达。结果与对照组比较,TS组脑细胞形态被破坏,干预1、2、3周小鼠运动行为评分、刻板行为评分及纹状体组织中NE、DA水平、TH阳性神经元数量高(P均<0.05),纹状体组织中5-HT水平及cAMP、PKA、CREB蛋白表达低(P均<0.05);与TS组比较,SS-L组、SS-H组、阳性对照组脑细胞形态得到改善,干预1、2、3周小鼠运动行为评分、刻板行为评分及纹状体组织中NE、DA水平、TH阳性神经元数量低(P均<0.05),纹状体组织中5-HT水平及cAMP、PKA、CREB蛋白表达高(P均<0.05);S-H+H-89组逆转SS-H组各指标的变化(P均<0.05)。结论SS可能通过调节cAMP/PKA/CREB信号通路对TS小鼠发挥治疗作用。Objective To investigate the therapeutic effect of saikosaponin(SS)on Tourette syndrome(TS)model mice based on cyclic adenosine monophosphate/protein kinase A/cAMP response element binding protein(cAMP/PKA/CREB)signaling pathway.Methods We randomly divided the mice into the control group,TS group,low-dose SS(SS-L)group,high-dose SS(SS-H)group,positive drug group,and SS-H+PKA inhibitor(H-89)group,with 10 mice in each group.Except for the control group,TS mouse models were established by intraperitoneal injection of iminodiacetonitrile solution in all other groups.After modeling,mice in the SS-L group and SS-H group were given intraperitoneal injection of 25 and 100 mg/kg SS,respectively,and gavaged with normal saline.Mice in the positive drug group were gavaged with 0.5 mg/kg haloperidol and intraperitoneally injected with normal saline.Mice in the SS-H+H-89 group were injected intraperitoneally with 100 mg/kg SS and 5 mg/kg H-89,and were given physiological saline by gavage.Mice in the TS group and control group were intraperitoneally injected with and gavaged with equal volume of normal saline,respectively,once a day,for three consecutive weeks.The motor behavior and stereotyped behavior of mice were scored,5-hydroxytryptamine(5-HT),norepinephrine(NE),and dopamine(DA)in striatal tissues were detected by ELISA,the morphological changes in the brain tissues were observed by hematoxylin eosin method,tyrosine hydroxylase(TH)positive neurons in substantia nigra were detected by the immunohistochemical method,and the cAMP/PKA/CREB pathway-related protein expression was detected by Western blotting.Results Compared with the control group,the cell morphology of the TS group was disrupted,at 1,2,and 3 weeks after intervention,the motor behavior,stereotypical behavior scores,the levels of NE and DA,and the number of TH positive neurons increased in mice,but the expression of 5-HT,cAMP,PKA,and CREB were reduced(all P<0.05).Compared with the TS group,the cell morphology of the SS-L group,SS-H group,and positive control grou

关 键 词：多发性抽动症 柴胡皂苷 环磷酸腺苷/蛋白激酶A/环磷酸腺苷反应成分结合蛋白信号通路 

分 类 号：R748[医药卫生—神经病学与精神病学]