黄芪甲苷Ⅳ对TGF-β1/Smad2介导的癫痫合并抑郁小鼠的保护作用  

Protective effect of astragaloside Ⅳ on TGF-β1/Smad2-mediated epilepsy combined with depression in mice

作　　者：罗湘蓝 刘平 张望 苏雯靓 王苏亚 阎乃宽[2] 陈建国[2] 苏华龙[1] LUO Xiang-lan;LIU Ping;ZHANG Wang;SU Wen-liang;WANG Su-ya;YAN Nai-kuan;CHEN Jian-guo;SU Hua-long(Chongqing Mental Health Center,Chongqing 400000,China;Geleshan Campus,Chongqing Mental Health Center,Chongqing 400036,China)

机构地区：[1]重庆市精神卫生中心,重庆400000 [2]重庆市精神卫生中心歌乐山院区,重庆400036

出　　处：《实用药物与临床》2021年第4期301-306,共6页Practical Pharmacy and Clinical Remedies

基　　金：重庆市精神卫生中心院级科研项目(2018-yjkt-05);重庆市精神卫生中心医学特色专科和新兴培育学科建设项目(渝精卫发<2018>50号)。

摘　　要：目的探讨黄芪甲苷Ⅳ对癫痫合并抑郁小鼠的治疗和保护作用及机制。方法 60只C57/BL6小鼠,随机分为对照组、癫痫组、黄芪甲苷Ⅳ低剂量组(20 mg/kg)、黄芪甲苷Ⅳ高剂量组(40 mg/kg)、黄芪甲苷Ⅳ+LY2109761组(40 mg/kg+1μg/只),每组12只。以氯化锂(180 mg/kg)联合匹罗卡品(50 mg/kg)制造小鼠癫痫合并抑郁模型,造模前7 d,除对照组和癫痫组,其余各组连续灌胃给药对应剂量的黄芪甲苷,造模前3 d,黄芪甲苷Ⅳ+LY2109761组小鼠给予LY2109761,1μg/只,鼻滴。造模结束后30~90 min,观察小鼠癫痫发作次数与程度,造模结束后24 h,检测小鼠糖水偏好,游泳不动时间及悬尾不动时间。行为学检测结束后取脑组织用于Nissl染色、Western blot实验。结果行为学测试显示,氯化锂联合匹罗卡品成功点燃小鼠癫痫症状,并造成小鼠抑郁样行为。黄芪甲苷Ⅳ预给药能够显著改善小鼠癫痫症状与抑郁样症状,而转化生长因子-β1(Transforming growth factor-β1,TGF-β1)抑制剂逆转了黄芪甲苷Ⅳ的治疗效果。此外,氯化锂联合匹罗卡品造成TGF-β1表达下降,其下游Smad2磷酸化水平降低,海马组织中Caspase-3剪切增加,而Bcl-2/Bax表达比例降低,最终造成小鼠海马组织中神经元凋亡。黄芪甲苷组TGF-β1蛋白表达显著升高,p-Smad2表达增加,Caspase-3蛋白剪切抑制,Bcl-2/Bax表达比例增加,神经元损伤减少。结论氯化锂联合匹罗卡品造成小鼠出现癫痫与抑郁样症状。黄芪甲苷Ⅳ通过增加TGF-β1表达,提高Smad2磷酸化水平,减少Cleaved-caspase-3的表达,抑制神经元凋亡,最终改善小鼠癫痫和抑郁样症状。Objective To explore the protective effect and mechanism of astragaloside Ⅳ on epilepsy combined with depression in mice.Methods Sixty C57/BL6 mice were randomly divided into control group, epilepsy group, ASG Ⅳ-L group(20 mg/kg),ASG Ⅳ-H group(40 mg/kg),ASG Ⅳ+LY2109761 group(40 mg/kg+1 μg per mouse),12 in each group.Lithium chloride(180 mg/kg) combined with pilocarpine(50 mg/kg) were administered to create the model of epilepsy with depression.Seven days before drug administration, except for the control group and the epilepsy group, the other groups were given the corresponding dose of ASG Ⅳ.Three days before drug administration, ASG Ⅳ + LY2109761 group received a nasal drip of 1 μg LY2109761 per mouse.The times and level of seizure in mice were observed in 30~90 min after the establishment of model, and the sugar preference, the immobility time of forced swimming and the suspansion were measured 24 h after the establishment of model.After the behavioral test, brain tissues were taken for Nissl staining and Western blot.Results Behavioral tests showed that lithium chloride combined with pilocarpine successfully brought about epileptic symptoms and depression-like behavior in mice.Pre-administration of ASG Ⅳ could significantly improve epilepsy and depression-like symptoms in mice, while TGF-β1 inhibitors reversed the effect of ASG Ⅳ.In addition, lithium chloride combined with pilocarpine caused a decrease in TGF-β1 expression and Smad2 phosphorylation level in the downstream, resulting in an increase in Cleaved-Caspase-3 in the hippocampus and a decrease in the Bcl-2/Bax expression rate, which ultimately caused neuronal apoptosis in hippocampus of mice.In the ASG group, TGF-β1 protein expression was significantly increased, p-Smad2 expression was increased, Cleaved-Caspase-3 protein was inhibited, Bcl-2/Bax expression rate was increased, and neuronal damage was reduced.Conclusion Lithium chloride combined with pilocarpine can cause epilepsy and depression-like symptoms in mice.ASG Ⅳ increa

关 键 词：癫痫 抑郁症 黄芪甲苷Ⅳ 神经元凋亡 

分 类 号：R285.5[医药卫生—中药学]