复方地黄颗粒对阴虚动风证帕金森病大鼠纹状体细胞凋亡及PI3K/Akt信号通路的影响

Effect of compound rehmannia granule on apoptosis of striatum cell and PI3K/Akt signal pathway in Parkinson's disease rats with endogenous wind syndrome caused by YIN Deficiency

目的探讨复方地黄颗粒(CRG)对阴虚动风证帕金森病(PD)大鼠纹状体细胞凋亡及对PI3K/Akt信号通路的影响。方法采用6-羟基多巴胺(6-OHDA)注射损毁黑质法制备阴虚动风证PD大鼠模型,造模成功30只大鼠。随机分为模型组、美多芭(MAD)组、复方地黄颗粒低(L-CRG)、中(M-CRG)、高(H-CRG)剂量组,每组6只。另纳入正常对照组和假手术组各6只。美多芭组予美多芭150 mg/(kg·d)灌胃; L-CRG、M-CRG、H-CRG组予复方地黄颗粒3. 5 g/(kg·d)、7 g/(kg·d)、14 g/(kg·d)灌胃;正常对照组、假手术组及模型组予等量生理盐水灌胃。用药6周后处死大鼠,取纹状体。采用TUNEL法检测纹状体细胞凋亡情况;采用透射电子显微镜观察纹状体细胞超微结构;采用蛋白印迹(WB)法检测Bcl-2、Bax、PI3K、Akt、p-Akt蛋白水平。结果与对照组和假手术组比较,模型组细胞凋亡指数明显升高,差异有统计学意义(P <0. 05),细胞凋亡特征明显,Bax蛋白水平明显升高(P <0. 05),Bcl-2、PI3K、p-Akt蛋白水平降低(P <0. 05);与模型组比较,低中高剂量复方地黄颗粒组细胞凋亡指数显著下降(P <0. 05),细胞形态趋于正常,Bax蛋白水平明显降低(P <0. 05),Bcl-2、PI3K、p-Akt蛋白水平升高(P <0. 05),且呈剂量依赖效应。结论复方地黄颗粒可能通过PI3K/Akt信号通路调节凋亡相关蛋白表达,抑制阴虚动风证PD大鼠纹状体细胞凋亡,呈剂量依赖效应。

Objective To investigate the effect of Compound Rehmannia Granule(CRG)on striatum cell apoptosis and PI3K/Akt signal pathway in Parkinson's disease(PD)rats with endogenous wind syndrome caused by YIN deficiency.Methods The model of PD rats with endogenous wind syndrome caused by YIN deficiency was prepared by injecting 6-hydroxydopamine(6-OHDA)for the damaging of the substantia nigra method,and 30 rats were successfully built.Divided them randomly into model group,Madopar group(MAD),Compound Rehmannia Granule low(L-CRG),Medium(M-CRG),high(H-CRG)dose group,6 rats in each group.Another 6 rats were also included in the normal control group and the sham operation group respectively.The Madopar group was give 150 mg/(kg·d)Madopar by intragastric administration,the L-CRG group,M-CRG group,H-CRG group were given 3.5 g/(kg·d),7 g/(kg·d),14 g/(kg·d)Compound Rehmannia Granule respectively by intragastric administration,the normal control group,the sham operation group and the model group were given the same amount of normal saline by intragastric administration.After 6 weeks of medication,the rats were killed and the striatum was taken.The apoptosis of striatum cells was detected by TUNEL;the ultrastructure of the striatum cells was observed by transmission electron microscope;the levels of Bcl-2,Bax,PI3K,Akt and p-Akt protein were detected by Western blot(WB).Results Compared with the control group and the sham operation group,the cell apoptosis index of the model group was significantly higher,and the difference was statistically significant(P<0.05).The characteristics of cell apoptosis were obvious,and the level of Bax protein was significantly increased(P<0.05),the levels of Bcl-2,PI3K and p-Akt protein were decreased(P<0.05).Compared with the model group,the cell apoptosis index of the L-CRG,M-CRG,H-CRG compound rehmannia granule group was significantly decreased(P<0.05).The cell morphology tended to normal,the level of Bax protein decreased significantly(P<0.05).The levels of Bcl-2,PI3K and p-Akt protein were increased(P<0.05)in a dose-dependent manner.Conclusion Compound Rehmannia Granule can inhibit the cell apoptosis of PD rats with endogenous wind syndrome caused by YIN deficiency in a dose-dependent manner.It may achieve by regulating the expression of apoptosis related proteins through PI3K/Akt signal pathway.