复方半枝莲通过调控自噬对肺癌大鼠氧化应激水平及解聚素金属蛋白酶10蛋白的作用机制

Action mechanism of compound scutellaria biloba on the level of oxidative stress and ADAM10 protein in lung cancer rats by regulating autophagy

目的探讨复方半枝莲通过调控自噬对肺癌大鼠氧化应激水平及解聚素金属蛋白酶10(ADAM10)蛋白的作用机制。方法将50只大鼠随机分为正常组(NC组),肺癌模型大鼠组(LC组),低剂量复方半枝莲组(CSB1组),中剂量复方半枝莲组(CSB2组),高剂量复方半枝莲组(CSB3组),每组10只。检测肿瘤体积和肿瘤抑制率,肺组织病理学变化,氧化应激指标;蛋白质印迹检测自噬相关蛋白LC3B、p62蛋白及ADAM10蛋白表达(P<0.05)。结果NC组大鼠没有肿瘤形成。LC组大鼠肿瘤体积明显增加,CSB1组、CSB2组和CSB3组大鼠肿瘤体积在治疗后均显著小于LC组(P<0.05)。和NC组相比,LC组大鼠氧化应激指标SOD活性和GSH-Px含量均显著降低(P<0.05),MDA含量明显升高(P<0.05);和LC组相比,CSB1组、CSB2组和CSB3组大鼠氧化应激指标SOD活性和GSH-Px含量均显著增加(P<0.05),MDA含量明显降低(P<0.05),且三组大鼠氧化应激指标变化成浓度依赖(P<0.05),和NC组相比,LC组ADAM10蛋白表达显著增加(P<0.05);和模型组相比,CSB1组、CSB2组和CSB3组中ADAM10蛋白表达均显著降低(P<0.05),CSB3组肺组织中ADAM10蛋白表达明显低于CSB1组、CSB2组(P<0.05)。自噬相关蛋白表达比较,和NC组相比,LC组中LC3B-Ⅱ/LC3B-Ι比值显著降低(P<0.05),p62蛋白表达明显升高(P<0.05);CSB1组、CSB2组和CSB3组中LC3B-Ⅱ/LC3B-Ι比值较LC组均不同程度升高(P<0.05),p62蛋白表达较LC组均显著降低(P<0.05);其中CSB3组中LC3B-Ⅱ/LC3B-Ι比值明显高于CSB1组、CSB2组(P<0.05),p62蛋白表达明显低于CSB1组、CSB2组(P<0.05)。结论复方半枝莲可通过调控自噬改善肺癌大鼠氧化应激水平,抑制大鼠组织中ADAM10表达,进而抑制肺癌发展。

Objective To explore the action mechanism of compound scutellaria barbata(CSB)on oxidative stress level and disintegrin and metalloproteinase 10(ADAM10)protein in lung cancer rats by regulating autophagy.Methods Fifty rats were randomly divided into normal control group(NC group),lung cancer(LC)model group,CSB1(low-dose)group,CSB2(middle-dose)group,CSB3(high-dose)group,10 in each group.Tumor volume and tumor inhibition rate,lung histopathological changes,oxidative stress indexes were detected.The expression of autophagy-related proteins LC3B,P62 and ADAM10 were detected by western blotting(WB).Results There was no tumor formation in NC group.Compared with NC group,the tumor volume in model group was significantly increased,while that was significantly lower in the CSB groups than in the model group after treatment(P<0.05).Compared with NC group,superoxide dismutase(SOD)activity and glutathione peroxidase(GSH-Px)level in model group were significantly decreased,while malondialdehyde(MDA)level was significantly increased(P<0.05).Compared with model group,the SOD activities and GSH-Px levels in CSB groups were significantly increased,while MDA levels were significantly decreased(P<0.05),and oxidative stress indexes in the CSB groups were concentration dependent(P<0.05).Compared with NC group,the protein expression of ADAM10 in model group were significantly increased(P<0.05).Compared with the model group,the protein expressions of ADAM10 in CSB groups were significantly decreased,and the protein expression of ADAM10 in lung tissues of CSB3 group was significantly lower than that of CSB1 and CSB2 groups(P<0.05).Compared with NC group,the ratio of LC3B-Ⅱ/LC3B-Ιin model group was significantly decreased,while the expression of p62 protein was significantly increased(P<0.05).Compared with the model group,the ratio of LC3B-Ⅱ/LC3B-Ιin CSB groups was significantly higher,and the expression of p62 protein was significantly lower(P<0.05).The LC3B-Ⅱ/LC3B-Ιin CSB3 group was significantly higher than that in CSB1 group and CSB2 group,while the protein expression of P62 was significantly lower(P<0.05).Conclusion CSB can improve the level of oxidative stress in LC rats by regulating autophagy,inhibit the expression of ADAM10 in lung tissues,and then inhibit the development of lung cancer.