黄连解毒汤对MPP+诱导的PC12细胞损伤的保护作用

 目的观察黄连解毒汤(HJD)对1-甲基-4-苯基-1,2,3,6-四氢吡啶离子(MPP⁺)所致大鼠嗜铬神经瘤细胞(PC12)损伤的保护作用并探讨其可能的作用机制。方法在生长状态良好的PC12细胞中加入终浓度为200μmol·L^-1的MPP⁺,造成帕金森病的离体实验模型。观察HJD全方,单味药及药对的大鼠含药血清对PC12细胞的保护作用。用四甲基偶氮唑盐(MTT)法检测细胞活力,用吖啶橙(AO)溴化乙锭(EB)荧光染色法观察细胞的形态改变,应用流式细胞仪检测细胞周期以及细胞线粒体膜电位。结果HJD的含药血清和单味药栀子的含药血清能明显提高MPP⁺损伤的PC12细胞活力,且全方的作用大于单味药栀子的作用。在200μmol·L^-1MPP⁺作用下,PC12细胞形态发生了凋亡和坏死,而HJD的含药血清能明显改善受损的细胞形态。流式细胞仪检测结果表明,HJD的含药血清能促进细胞从G₁期向S期和G₂期的过度,并能对抗MPP⁺所致的PC12细胞的线粒体膜电位的降低。结论HJD对MPP⁺所致的PC12细胞的损伤具有保护作用,它能减少细胞的凋亡和坏死,提高细胞存活率。调节细胞周期和维护线粒体膜电位可能是HJD保护受损PC12细胞的作用机制之一。

Protective Effect of Huanglian Jiedu Decoction on the Injury of PC12 Cells Induced by MPP+

OBJECTIVE To observe the protective effect of Huanglian jiedu decoction(HJD)on the PC12 cell injury induced by 1-methyl-4-phenylpyridinium(MPP~+)and study the mechanism.METHODS Cell model of Parkinson's disease was established by adding 200μmol·L^-1MPP⁺ into PC12 cell.The protective effects of rat serum concentrating HJD or its components on PC12 cell were assessed.The cell viability was analyzed by methyl thiazolyl tetrazolium(MTT)method,the morphological changes were observed by acridine orange(AO)and ethidium bromide(EB)staining assay.Cell cycle and mitochondrial transmembrane potential were meas- ured by flow cytometry.RESULTS The serum concentrating HJD or Gardenia jasminoides Ellis can significant enhance PC12 cell's viability,the effect of former was stronger.Under 200μmol·L^-1MPP⁺,the morphological of PC12 cell was changed,including ap- optosis and necrosis.The serum concentrating HJD can notable protect these injuries of PC12 cell.Using flow cytometry,we can ob- serve the serum concentrating HJD can decrease the percentage of G1 phase and increase the percentage of S phase and G2 phase.The protection on the mitochondrial membrane potential of PC12 was also observed.CONCLUSION HJD has the potential to protect PC12 cell which injured by MPP⁺.It can decrease the cell apoptosis and necrosis,raising the cell viability.Regulating the cell cycle and maintenancing the mitochondrial transmembrane potential may be the possible mechanism of HJD's protection.