左归丸含药血清对大鼠海马神经元和胶质细胞氧糖剥夺损伤的保护作用及机制研究

Protective effect and mechanism of Zuogui pill-containing serum on oxygen and glucose deprivation injury of hippocampal neurons and glial cells in rats

目的:探讨左归丸含药血清对大鼠海马神经元和胶质细胞氧糖剥夺损伤的保护作用及机制。方法:原代培养大鼠海马神经元和胶质细胞混合细胞,分为对照组、模型组、含药血清组和抑制剂组。对照组细胞不做特别处理。模型组细胞给予氧糖剥夺。含药血清组细胞除给予氧糖剥夺+左归丸含药血清。抑制剂组细胞给予氧糖剥夺+左归丸含药血清+雌激素受体(ER)非特异性拮抗剂ICI182780。采用CCK-8法检测各组细胞存活率。采用酶联免疫吸附(ELISA)法检测各组细胞氧化应激和炎症因子谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)表达水平。采用Western blot检测TNF-α、IL-1β、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关x蛋白(Bax)、半胱氨酸蛋白酶-3(Caspase-3)及磷脂酰肌醇激酶(PI3K)/蛋白激酶B(AKT)通路相关蛋白表达水平。结果:与对照组比较,模型组细胞存活率、GSH-Px水平下降,MDA、TNF-α、IL-1β水平升高,Bcl-2、磷酸化磷脂酰肌醇激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)蛋白表达下降,Bax、Caspase-3蛋白表达增加(均P<0.05)。与模型组比较,含药血清组细胞存活率、GSH-Px水平增加,MDA、TNF-α、IL-1β水平下降,Bcl-2、p-PI3K、p-AKT蛋白表达显著上调,Bax、Caspase-3蛋白表达显著下调(均P<0.05)。结论:左归丸含药血清对大鼠海马神经元和胶质细胞氧糖剥夺损伤具有保护作用,其机制与调控氧化应激、炎性反应及PI3K/AKT通路有关。

Objective:To investigate the protective effect and mechanism of Zuogui pill-containing serum on oxygen and glucose deprivation injury of hippocampal neurons and glial cells in rats.Methods:The mixed cells of hippocampal neurons and glial cells were primarily cultured and divided into control group,model group,drug containing serum group and inhibitor group.The cells in the control group were not specially treated.The cells in the model group were given oxygen and glucose deprivation.The cells in the drug containing serum group were given oxygen and glucose deprivation+Zuogui pill-containing serum.The cells in the inhibitor group were given oxygen and glucose deprivation+Zuogui pill-containing serum+estrogen receptor(ER)nonspecific antagonist ICI182780.The cell survival rate of each group was detected by CCK-8 method.The expression levels of oxidative stress and inflammatory factors such as GSH-Px,MDA,TNF-α,and IL-1βwere detected by ELISA.The expression levels of TNF-α,IL-1β,Bcl-2,Bax,Caspase-3 and PI3K/AKT pathway related proteins were detected by Western blot.Results:Compared with the control group,in the model group,the cell survival rate and GSH-Px level decreased,the levels of MDA,TNF-αand IL-1βincreased,the levels of Bcl-2,p-PI3K and p-AKT proteins decreased,the levels of Bax and Caspase-3 proteins increased(all P<0.05).Compared with the model group,in the drug containing serum group,the cell survival rate and GSH-Px level increased,the levels of MDA,TNF-αand IL-1βdecreased,the levels of Bcl-2,p-PI3K and p-AKT proteins increased,the levels of Bax and Caspase-3 proteins decreased(all P<0.05).Conclusion:Zuogui pill-containing serum has a protective effect on oxygen and glucose deprivation injury of hippocampal neurons and glial cells in rats,and its mechanism is related to the regulation of oxidative stress,inflammatory response and PI3K/AKT pathway.