补阳还五精简方的体外筛选及体内验证研究

摘要

Objective To screen the optimized Buyang Huanwu Decoction (BYHWD);To verify it. Methods H2O2 was used to induce PC12 cell oxidative stress models. MTT method was used to determine the prevention effects of BYHWD at different concentrations (0.1, 0.2, 0.5, 1.0, 2.0, 3.5 mg/mL) on in vitro oxidative stress cell models to define the optimized concentration. Orthogonal design was used to divide BYHWD single medicine into decomposed BYHWD groups, control group (only with DMEM), normal group (without H2O2 and medicine processing), and model group, to investigate the protective effects on PC12 cells. Optimized BYHWD was screened to decide the compatibility ratio of each medicine. MTT was used to detect the cell survival rate in each group. Middle cerebral artery occlusion was used to replicate MACO rat models. SD rats were randomly divided into sham-operation group, model group, BYHWD group and optimized BYHWD high-, medium-and low-dose groups. Each medication group was given relevant medicine for gavage. The screened results were verified. Results Compared with other decomposed BYHWD groups, the protective effects of the compatibility of Astragali Radix+Chuanxiong Rhizoma+Pheretima on PC12 cells was the best (P<0.05), which was nearly equaled to BYHWD. Compared with the model group, BYHWD and the optimized one could evidently reduce cerebral cortex infarction area and improve the impaired brain edema (P<0.05), and the medium-dose group was the best. Conclusion The optimized BYHWD ratio is:Astragali Radix:Chuanxiong Rhizoma:Pheretima=10:3:1.%目的：筛选补阳还五精简方的最佳配伍，并进行验证。方法采用H2O2诱导建立PC12细胞氧化应激模型。考察不同浓度0.1、0.2、0.5、1.0、2.0、3.5 mg/mL补阳还五汤对体外氧化应激模型细胞的保护作用，确定最佳作用浓度。采用正交试验设计，将补阳还五汤各单味药随机配伍分成补阳还五汤不同拆方组及对照组（仅加细胞培养基）、正常组（无H2O2和药物处理）、模型组,考察对PC12细胞的保护作用，筛选精简方，确定精简方各单味药的配伍比例，MTT 法测定各组细胞存活率。采用大脑中动脉线栓法复制局灶性脑缺血模型，SD大鼠随机分为假手术组、模型组、补阳还五汤组和精简方高、中、低剂量组，各给药组给予相应药物灌胃，验证筛选结果。结果与其他拆方组比较，黄芪＋川芎＋地龙对PC12细胞的保护作用最佳（P＜0.05），且与补阳还五汤原方相当；与模型组比较，补阳还五汤组及精简方各剂量组均能改善模型大鼠大脑皮层梗死面积、降低脑组织水肿程度（P＜0.05），且以精简方中剂量组最佳。结论补阳还五精简方最佳配伍为黄芪、川芎、地龙，其比例为10∶3∶1。