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目的:探讨姜黄素对大鼠气道平滑肌细胞(airway smooth muscle cells,ASMCs)增殖和凋亡的影响。方法:采用改良组织块消化法培养原代大鼠气道平滑肌细胞,以PDGF诱导ASMCs增殖建立模型。MTT法检测不同浓度姜黄素抑制ASMCs增殖情况,Hoechst33342染色和DNA Ladder检测细胞凋亡,Western Blot检测ERK1/2和磷酸化ERK1/2的表达。结果:①MTT检测给予姜黄素处理12h后,与模型组相比较,10μmol/l组、20μmol/l组和40μmol/l组的细胞平均抑制率均增加显著,P<0.05;48h后各浓度组抑制率均升高。②Hoechst33342观察到10μmol/l、20μmol/l和40μmol/l姜黄素组中强荧光细胞比例随姜黄素剂量增大而增多,细胞核内多个不均一蓝染现象。③DNALadder观察到40μmol/l组姜黄素处理组出现梯状分布。④姜黄素(40μmol/l)与PDGF(20ng/ml)共同处理30min和60min后p-ERK1/2蛋白表达水平显著降低。结论:姜黄素对ASMCs增殖有抑制作用,同时高浓度的姜黄素可促进ASMCs凋亡,可能与下调ERK1/2的表达有关。
Objective: To investigate the effects of curcumin on proliferation and apoptosis of airway smooth muscle cells (ASMCs) in rats. Methods: Primary rat airway smooth muscle cells were cultured with modified tissue block digestion method, and the proliferation of ASMCs was induced by PDGF. MTT assay was used to detect the proliferation of ASMCs with different concentrations of curcumin. Hoechst33342 staining and DNA Ladder were used to detect the apoptosis. The expressions of ERK1 / 2 and phosphorylated ERK1 / 2 were detected by Western Blot. Results: (1) MTT assay showed that the average inhibitory rates of cells in 10μmol / l group, 20μmol / l group and 40μmol / l group increased significantly after treated with curcumin for 12h, P <0.05; Rates are higher. ②Hoechst33342 observed that the proportion of strong fluorescent cells in 10μmol / l, 20μmol / l and 40μmol / l curcumin groups increased with the increase of curcumin dose, and there were many non-uniform blue staining in the nucleus. ③ DNALadder observed 40μmol / l group curcumin treatment group appeared ladder-like distribution. ④ The expression of p-ERK1 / 2 in curcumin (40μmol / l) and PDGF (20ng / ml) for 30min and 60min significantly decreased. CONCLUSION: Curcumin can inhibit the proliferation of ASMCs. At the same time, high concentrations of curcumin can promote the apoptosis of ASMCs, which may be related to the down-regulation of ERK1 / 2 expression.