补阳还五汤含药血清对缺氧缺糖损伤PC12细胞凋亡影响的正交试验研究

摘要

【目的】观察补阳还五汤对缺氧缺糖（oxygen-glucose deprivation， OGD）损伤PC12细胞凋亡的影响，筛选补阳还五汤中抑制OGD损伤PC12细胞凋亡的主要药味。【方法】将补阳还五汤中7种单味药即黄芪、赤芍、当归尾、地龙、川芎、桃仁、红花作为实验因素，每个因素选取有或无2个水平，按L8（27）正交试验表安排试验。采用OGD细胞模型，运用中药血清药理学方法和流式细胞术技术，以细胞凋亡率为评价指标，利用直观分析和方差分析法，比较空白血清组（有氧+含糖培养基+空白血清）、模型组（缺氧+无糖培养基+空白血清）和补阳还五汤（缺氧+无糖培养基+含药血清）以及各拆方对OGD损伤PC12细胞凋亡的影响。【结果】与空白血清组比较，模型组PC12细胞凋亡率显著增加，差异有统计学意义(P<0．01)；与模型组比较，补阳还五汤组的凋亡率显著降低，差异有统计学意义(P<0．01)。正交试验分析显示：补阳还五汤中对OGD损伤PC12细胞凋亡具有显著性影响的药味为桃仁、红花、赤芍(P<0．01)，其余药味作用不显著(P>0．05)，含赤芍的拆方细胞凋亡率低于不含赤芍的拆方。【结论】补阳还五汤具有抑制OGD损伤PC12细胞凋亡的作用，其中发挥作用的主要药味为赤芍。%Objective To investigate the protective effect of Buyang Huanwu Decoction ( BYHWD) on PC12 cell apoptosis induced by oxygen-glucose deprivation ( OGD) and to screen out the main ingredients of BYHWD on protecting PC12 cell from injury induced by OGD. Methods Seven herbs of BYHWD were selected as the influencing factors, and they were Radix Astragali, Radix Paeoniae Rubra, the carda part of Radix Angelicae Sinensis, Pheretima Asiatica, Rhizoma Chuanxiong, Semen Persicae, and Flos carthami. Two levels were designed for each factor, and the experiment was arranged according to L8 ( 27) orthogonal test table. Apoptosis of PC12 cells induced by OGD was detected by serum pharmacology method and flow cytometry techniques. With apoptotic rate as the evaluation index, visual analysis and variance analysis were used for the comparison of apoptosis of PC12 cells in the blank control group ( oxygen + sugary medium + blank serum) , model group (oxygen-glucose-free medium + blank serum) and drug groups (oxygen-glucose-free medium +serum containing BYHWD or the separate recipe of BYHWD ) . Results Compared with the blank control group, apoptotic rate was increased in PC12 cells of the model group, the difference was statistically significant ( P<0.01) . Compared with the model group, the apoptotic rate was decreased in BYHWD group, and the differences was statistically significant (P<0.01) . The results of orthogonal analysis showed that Semen Persicae, Flos carthami and Radix Paeoniae Rubra were the main active ingredients of BYHWD on depressing apoptosis of PC12 cell induced by OGD ( P<0.01). The apoptotic rate of serum containing the prescription with Radix Paeoniae Rubra was lower than that of the prescription without Radix Paeoniae Rubra. Conclusion BYHWD could inhibit OGD-induced PC12 cell apoptosis, and Radix Paeoniae Rubra is the main active herb on depressing apoptosis in BYHWD.