复方地黄颗粒对阴虚动风证帕金森病模型大鼠神经营养因子及其受体的影响

The compound Rehmannia granule upregulates neurotrophic factors and their receptors in the rat model of Parkinson's disease with endogenous wind syndrome caused by YIN deficiency

目的探讨复方地黄颗粒治疗阴虚动风证帕金森病(PD)的作用机制,为临床PD治疗提供新的思路。方法制备6-羟基多巴胺(6-OHDA)阴虚动风证PD大鼠模型,将造模成功的50只大鼠随机分为模型组,美多芭组和复方地黄颗粒低、中、高剂量组;另取不造模大鼠10只不作处理设为正常对照组,10只只注射抗坏血酸设为假手术组。美多芭组给予美多芭150mg/kg,复方地黄颗粒低、中、高剂量组分别给颗粒剂3.5、7、14g/kg灌胃;正常对照组、假手术组和模型组以等体积蒸馏水(2ml/只)灌胃,每日1次,连续用药6周。分别采用RT-q PCR及Western blotting检测神经生长因子及其受体(NGF)、脑源性神经营养因子(BDNF)、胶质细胞源性神经营养因子(GDNF)、原肌球蛋白受体激酶(Trk)A、Trk B、GDNF家族受体(GFR)-α1以及Ret的m RNA及蛋白在损毁侧纹状体的表达,酪氨酸羟化酶(TH)m RNA及蛋白在损毁侧黑质及纹状体的表达。结果与正常对照组比较,模型组大鼠NGF、BDNF、GDNF、Trk A、Trk B、GFR-α1、Ret、TH的m RNA及蛋白表达均明显降低(P<0.01)。与模型组比较,美多芭组以及复方地黄颗粒各剂量组神经营养因子及其受体的表达均明显升高(P<0.01),但仅中剂量组表达水平与美多芭组比较差异无统计学意义(P>0.05)。结论复方地黄颗粒可能通过调控神经营养因子及其受体的表达来发挥抗PD作用,其中以中剂量复方地黄颗粒的疗效为佳。

Objective To investigate the mechanism of the compound Rehmannia granule in treating Parkinson disease （PD） of endogenous wind syndrome caused by deficiency of YIN, and to provide potential treatments to the Parkinson＇s disease. Methods To establish PD rat model with endogenous wind syndrome caused by deficiency of YIN, rats were treated with 6-Hydroxy-dopamine （6-OHDA）. A total of 50 rats that showed symptoms of PD were randomly divided into model group, Madopar group, and compound Rehmannia granule group （low, middle, high dose）. Ten rats without modeling were used as the normal control （NC） and rats received ascorbic acid were used as sham operation （SO） control. Madopar was given at a dosage of 150mg/kg. Compound Rehmannia granule was given at 3.5, 7, 14g/kg as low, middle and high dose. Both Madopar and compound Rehmannia granule was given using oral gavage. The NC group, SO group and model group were oral gavaged with the same volume of distilled water （2ml/rat） at a frequency of 1 times a day, for 6 weeks. The expression of levels of nerve growth factor （NGF）, brain derived neurotrophic factor （BDNF）, glial cell line-derived neurotrophic factor （GDNF）, tropomyosin receptor kinase A （TrkA）, TrkB, GDNF family receptor α1（GFR-α1）, RET in the damage side striatum and the expression levels of TH from the damage side substantia nigra striatum and striatum were quantified using RT-qPCR, Western blotting. Their expressions were further verified using immunofluorescence. The content of MDA, GSH,GSH-Px, and SOD in the damage side substantia nigrachemical were detected using the chemical colorimetric method. Results Comparing to the NC group, the mRNA and protein expression of NGF, BDNF, GDNF, TrkA,TrkB, GFR-α1, Ret, TH decreased in model group （P〈0.01）. Comparing to model rats, PD rats that were treated with Madopar and compound Rehmannia granule showed higher expression of neurotrophic genes （P〈0.01）. There was no statistic difference between Madopar group and compound Rehmannia granule middle dosage group （P〉0.05）. Conclusion Compound Rehmannia granule may play a role in anti Parkinson disease by improving the expression of neurotrophic factors and receptors.