姬松茸多糖酶降解对D-半乳糖诱导3T3细胞氧化损伤的保护作用

Enzymatic Degradation of Agaricus blazei and Its Protective Effect on Oxidative Damage Induced by D-gal in 3T3 Cells

目的优化并建立姬松茸多糖(Agaricus blazei polysaccharide,ABP)的酶解工艺,探讨其对D-半乳糖诱导小鼠胚胎成纤维细胞(3T3)衰老的保护作用.方法采用响应面法检测姬松茸多糖最佳酶解工艺;用D-半乳糖诱导氧化损伤;采用噻唑蓝(MTT)法检测各时期细胞活力;测定抗氧化指标.结果姬松茸多糖最佳酶解工艺:加酶量为2.9%,温度为59℃,时间为2.1 h,该酶解方法姬松茸多糖提取率为11.18%.选取培养48 h的3T3细胞进行自由基活性检测,其中模型组与对照组比较细胞存活率降低,差异具有统计学意义(P<0.01).给药组与对照组比较细胞存活率增加,在给药终浓度为100μg/mL时,细胞恢复效果最佳,细胞内SOD活力升高(P<0.05),CAT活力降低(P<0.01),MDA含量显著减少(P<0.01),差异具有统计学意义,并且呈现剂量依赖性.结论该方法建立了高效、稳定的姬松茸多糖酶解工艺,获得的姬松茸多糖能够抑制细胞氧化损伤,对D-半乳糖诱导的3T3细胞衰老具有保护作用.

Objective To optimize and establish the enzymatic hydrolysis process of Agaricus blazei polysaccharide(ABP)and explore its protective effect on D-galactose-induced mouse embryonic fibroblast(3T3)senescence.Method The optimal enzymatic hydrolysis process of Agaricus blazei Murill polysaccharide was detected by response surface methodology.The oxidative damage induced by D-galactose was induced.The cell viability was detected by MTT assay.The antioxidant index was determined.Results The optimum enzymatic hydrolysis process of Agaricus blazei polysaccharide was as follows:the enzyme dosage was 2.9%,the temperature was 59℃,and the time was 2.1 h.The extraction rate of Agaricus blazei polysaccharide was 11.1 8%.3T3 cells cultured for 48 hours were selected for free radical activity detection.Compared with the control group,the cell survival rate of the model group was decreased,which was statistically significant(P<0.0 1).Compared with the control group,the cell survival rate increased in the drug-administered group.At the final concentration of 100μg/mL,the cell recovery effect was the best,which could increase the intracellular SOD activity(P<0.05)and decrease the CAT activity(P<0.01),MDA content was significantly reduced(P<0.01),and both were statistically different and presented in a dose-dependent manner.Concl usion The high-efficiency and stable enzymatic hydrolysis process of Agaricus blazei Murill were established.Agaricus blazei polysaccharide obtained by this method can inhibit oxidative damage and protect 3T3 cell senescence induced by D-galactose.