绞股蓝乙醇提取物通过抑制细胞凋亡改善小鼠帕金森样行为  

Gynostemma pentaphyllum ethanol extract ameliorates motor dysfunction in a Parkinson’s disease mouse model through inhibiting neuronal apoptosis

作　　者：赵婷婷 何岚峤 严森 樊鹏雨 张翀 曾玲晖 ZHAO Tingting;HE Lanqiao;YAN Sen;FAN Pengyu;ZHANG Chong;ZENG Linghui(School of Medicine,Hangzhou City University,Zhejiang Provincial Key Laboratory of Novel Targets and Drug Study for Neural Repair,Hangzhou 310015,China;Institute of Brain and Cognitive Science,Hangzhou City University,Hangzhou 310015,China)

机构地区：[1]浙大城市学院医学院,浙江省神经损伤修复新靶点及药物研究重点实验室,浙江杭州310015 [2]浙大城市学院脑与认知研究院,浙江杭州310015

出　　处：《浙江大学学报(医学版)》2025年第1期49-57,共9页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省自然科学基金(LTGD23C090001);国家自然科学基金(31800888);浙大城市学院科研培育基金(X-202104,X-202304)。

摘　　要：目的:探究绞股蓝乙醇提取物(本文简称GP)对6-羟基多巴胺诱导的帕金森病小鼠的保护作用及潜在机制。方法:将C57BL/6雄性小鼠随机分为空白对照组、模型对照组、阳性对照组、GP小剂量组和GP大剂量组,每组16只。通过在小鼠黑质网状部区域定位注射6-羟基多巴胺的方式建立帕金森病小鼠模型。造模两周后,阳性对照组、GP小剂量组和GP大剂量组分别给予左旋多巴(10 mg/kg,腹腔注射)、GP(100 mg·kg^(-1)·d^(-1),口服)和GP(200 mg·kg^(-1)·d^(-1),口服)三周。通过旷场实验和CatWalk步态分析实验评估GP对帕金森病小鼠运动功能障碍的影响;通过抓力仪检测小鼠的肌肉力量;通过免疫荧光染色检测酪氨酸羟化酶(TH)阳性神经元数变化;通过酶联免疫吸附试验检测脑内多巴胺和血清素水平;通过蛋白质印迹法分析GP对磷酸化胞外信号调节激酶(ERK)1/2、磷酸化p38、磷酸化c-Jun氨基端激酶(JNK)等丝裂原活化蛋白激酶(MAPK)家族蛋白和B细胞淋巴瘤蛋白(Bcl)-2、Bcl-2关联X蛋白(Bax)、cleaved-胱天蛋白酶(caspase)-3等线粒体凋亡相关蛋白的影响。结果:行为学实验结果显示,与模型对照组比较,GP各剂量组移动距离、正常步序比、四肢步周长度增加,前肢抓力增大(均P<0.05)。进一步检测发现,与模型对照组比较,GP组TH阳性细胞比例增加,脑组织中多巴胺和血清素含量增加,磷酸化ERK1/2表达增加,磷酸化p38和磷酸化JNK1/2表达减少,Bax/Bcl-2和cleaved-caspase-3/caspase-3下降(均P<0.05)。结论:GP可通过调节磷酸化MAPK家族和线粒体凋亡相关蛋白的表达提高中脑组织内多巴胺和血清素水平,促进黑质网状部区域多巴胺能神经元存活,改善帕金森病小鼠的运动障碍。Objective:To investigate the protective effects and underlying mechanisms of Gynostemma pentaphyllum(GP)ethanol extract on motor dysfunction in a mouse model of Parkinson’s disease(PD).Methods:Eighty C57BL/6 male mice were randomly divided into five groups:control group,model group,levodopa group(positive control group),lowdose GP group,and high-dose GP group,with 16 mice per group.The PD model was induced by injection of 6-hydroxydopamine into the substantia nigra pars reticulata of the mice.Two weeks after 6-hydroxydopamine,positive control group received intraperitoneal injection of levodopa 10 mg·kg^(-1)·d^(-1),while low-dose GP and high-dose GP groups received GP extract 100 or 200 mg·kg^(-1)·d^(-1) orally for three weeks.After a 3-weektreatment,the effects of GP on motor dysfunction in 6-hydroxydopamine-induced PD were assessed using open field and CatWalk gait tests,while the effects on muscle strength were evaluated by forelimb grip strength.Immunofluorescence staining was used to detect the number of tyrosine hydroxylase(TH)positive neurons.The levels of dopamine and serotonin in the midbrain were determined by enzyme-linked immunosorbent assay.In addition,Western blotting was performed to detect the expression of mitogen-activated protein kinase(MAPK)family proteins such as p-extracellular signal-regulated kinase(ERK)1/2,p-p38 and p-c-Jun N-terminal kinase(JNK)1/2,and mitochondrial apoptosis pathway proteins such as B-cell lymphoma(Bcl)-2,Bcl-2 associated X protein(Bax),and cleaved-cysteine aspartic acid specific protease(caspase)-3.Results:Behavioral experiments showed that GP significantly improved the spontaneous activity and motor coordination of PD mice(P<0.05).The forelimb grip strength was also increased by GP treatment(P<0.05),compared to the PD model group.In addition,compared with the model group,the number of TH-positive neurons in substantia nigra pars reticulata region,the levels of dopamine and serotonin in midbrain and the expression of p-ERK1/2 were significantly increased by GP tr

关 键 词：帕金森病 绞股蓝 行为学实验 多巴胺能神经元 丝裂原活化蛋白激酶家族蛋白 线粒体凋亡相关蛋白 小鼠 

分 类 号：R285[医药卫生—中药学]