N-乙酰-L-半胱氨酸对子宫内膜异位症自噬及活性氧的影响

Effect of N-acetyl-L-cysteine on autophagy and reactive oxygen species in endometriosis

目的观察抗氧化剂N-乙酰-L-半胱氨酸(NAC)对子宫内膜异位症(EMs)异位病灶中活性氧(ROS)及自噬的影响,探讨其作用机制。方法将雌性成年Wistar大鼠随机分为空白组、假手术组、模型组、对照组和实验组,每组10只。空白组不予处理,假手术组开腹探查并关腹,模型组、对照组和实验组用EMs腹腔种植法造模。造模成功后,模型组无药物处理,对照组给予氯喹40mg·kg-1,腹腔灌注,实验组给予NAC 200 mg·kg-1,腹腔灌注。以免疫荧光法和蛋白质印迹(Western blot)法观察各组处理前后微管相关蛋白3-Ⅱ(LC3-Ⅱ)荧光斑点表达和蛋白水平情况,以酶联免疫吸附(ELISA)法检测各组ROS变化情况。结果模型组标本细胞内LC3-Ⅱ荧光表达密度增加,蛋白灰度明显升高(P <0. 05);对照组和实验组与空白组相比,LC3-Ⅱ荧光密度差异无统计学意义(P> 0. 05)。对照组和实验组标本细胞内ROS表达分别为102. 10±6. 03,97. 36±5. 17,与空白组(87. 75±4. 61)相比,差异均无统计学意义(均P> 0. 05)。结论抗氧化剂NAC可能降解EMs异位病灶中ROS,并抑制细胞自噬。

Objective To observe the effect of antioxidant N-acetyl-L-cysteine(NAC)on autophagy and reactive oxygen species(ROS)in ectopia lesions of endometriosis and its mechanism.Methods Fifty adult female Wistar rats were randomly divided into blank group,sham-operation group,model group,control group and experimental group,10 rats in each group.Blank group was given no treatment,sham operation group was opened for abdominal exploration and then closed,model group,control group and experimental group were modeled by EMs peritoneal implantation.After the animal model successfully made,model group was given no treatment,experimental group was given NAC 200 mg·kg-1 peritoneal perfusion,control group was given chloroquine 40 mg·kg-1 peritoneal perfusion.The express of light chain protein 3-Ⅱ(LC3-Ⅱ)fluorescence spots and protein were observed by immunofluorescence and Western blot.The enzyme-linked immunosorbent assay(ELISA)was use to obersve the ROS in each group.Results Density of LC3-Ⅱfluorescence spots and the gray intensity of LC3-Ⅱin model group increased obviously(P<0.05);the density of LC3-Ⅱfluorescence spots in control group and experimental group had no statistics difference compared with blank group(P>0.05).The expression of ROS in control and experimental groups were 102.10±6.03,97.36±5.17,had no significant difference compared with blank group,which was 87.75±4.61(P>0.05).Conclusion Antioxidant NAC may degraded ROS and inhibited autophagy in ectopia lesions of endometriosis.