补肺健脾方含药血清对香烟烟雾提取物诱导L6细胞中ROS、MnSOD和CAT表达的影响

Effects of Serum Containing Bufei Jianpi Recipe(补肺健脾方) on Expression of ROS,MnSOD and CAT in L6 Cells Induced by Cigarette Smoke Extract

目的探讨补肺健脾方治疗慢性阻塞性肺疾病(COPD)骨骼肌功能障碍的可能作用机制。方法50只大鼠分为空白对照组、补肺健脾方组各25只。空白对照组大鼠给予生理盐水2 ml/只灌胃,补肺健脾方组给予补肺健脾方悬乳液4. 51 g/(kg·d)的10倍剂量灌胃,每日1次。于第7日灌胃后1 h腹主动脉取血制备含药血清。正常培养的L6细胞分为正常组(10%空白对照组血清)、模型组[10%空白对照组血清+10%香烟烟雾提取物(CSE)]、补肺健脾组(10%补肺健脾方组血清+10%CSE)、西药组[10%空白对照组血清+10%CSE+5 mmol/L抗氧化剂N-乙酰半胱氨酸(NAC)]、补肺健脾+西药组(10%补肺健脾方组血清+10%CSE+5 mmol/L NAC)分别加入相应药物后,培养24 h。检测各组细胞中活性氧(ROS)水平及锰超氧化物歧化酶(MnSOD)、过氧化氢酶(CAT) mRNA和蛋白表达。结果模型组细胞中ROS水平、Mn SOD和CAT mRNA和蛋白表达较正常组明显升高(P <0. 05或P <0. 01),补肺健脾组、西药组和补肺健脾+西药组ROS水平、Mn SOD mRNA和蛋白及补肺健脾组CAT mRNA的表达较模型组降低(P <0. 05或P <0. 01)。与补肺健脾组比较,西药组和补肺健脾+西药组MnSOD和CAT mRNA表达下降更为明显(P <0. 05或P <0. 01)。结论补肺健脾方和NAC均能降低L6细胞中ROS水平,减轻氧化应激,且两者联合运用效果更佳,这可能是其改善COPD骨骼肌功能障碍的机制之一。

Objective To investigate the possible mechanism of Bufei Jianpi Recipe(补肺健脾方)in the treatment of skeletal muscle dysfunction in chronic obstructive pulmonary disease(COPD).Methods Fifty rats were divided into blank control group and Bufei Jianpi Recipe group,with 25 rats in each group.Rats in the blank control group were given normal saline 2 ml/rat by gavage,and the Bufei Jianpi Recipe group was given 10 times Bufei Jianpi Recipe suspension 4.51 g/(kg·d)by gavage,once a day.On the 7 th day,1 hour after intragastric administration,blood was taken from the abdominal aorta to prepare drug-containing serum.Normal cultured L6 cells were divided into normal group(10%blank control serum),model group[10%blank control group serum+10%cigarette smoke extract(CSE)],the Bufei Jianpi Recipe group(10%Bufei Jianpi Recipe serum+10%CSE),the western medicine group[10%blank control group serum+10%CSE+5 mmol/L antioxidant N-acetylcysteine(NAC)],the Bufei Jianpi Recipe+western medicine group(10%Bufei Jianpi Recipe serum+10%CSE+5 mmol/L NAC)were added to the corresponding drugs and cultured for 24 h.The levels of reactive oxygen species(ROS),manganese superoxide dismutase(MnSOD)and catalase(CAT)mRNA and protein were detected in each group.Results The levels of ROS,MnSOD and CAT mRNA and protein in the model group were significantly higher than those in the normal group(P<0.05 or P<0.01).The levels of ROS,MnSOD mRNA and protein in the Bufei Jianpi Recipe group,the western medicine group and Bufei Jianpi Recipe+western medicine group,and the expression of CAT mRNA in the Bufei Jianpi Recipe group were decreased than those in the model group(P<0.05 or P<0.01).Compared with the Bufei Jianpi Recipe group,the expression of MnSOD and CAT mRNA in the western medicine group and the Bufei Jianpi Recipe+western medicine group decreased more significantly(P<0.05 or P<0.01).Conclusion Bufei Jianpi Recipe Both Bufei Jianpi Recipe and NAC can reduce ROS levels in L6 cells and reduce oxidative stress,and the combined use achieves better effects,which may be one of the mechanisms to improve skeletal muscle dysfunction in COPD.