褪黑素对脂多糖诱导心肌肥厚的保护作用

发布时间：2019-05-22 17:56

【摘要】：目的研究探讨褪黑素对脂多糖(LPS)诱导心肌肥厚的保护作用及机制。方法 (1)培养的心肌细胞分为五组:对照组、LPS组、LPS+褪黑素(1.5mg/L)组、LPS+褪黑素(3mg/L)组、LPS+褪黑素(6mg/L)组,在倒置相差显微镜下观察心肌细胞形态学改变;(2)通过RC DCTM Protein Assay Kit检测各组心肌细胞蛋白水平。(3)通过酶联免疫吸附试验(ELISA)检测各组心肌细胞肿瘤坏死因子(TNF-α)的分泌水平;(4)利用Realtime PCR和Western blot检测各组心肌细胞肿瘤坏死因子受体1(TNFR1)基因和蛋白表达水平变化;(5)采用Ca2+探针Fura-2/AM在影像系统下记录各组心肌细胞Ca2+内流情况。(6)Western blot检测各组心肌细胞内CaN和CaMKⅡδB蛋白水平变化。结果 LPS能够诱导心肌肥厚、促进TNF-a的分泌和TNFR1的表达及增加Ca2+内流。在LPS诱导损伤之前预先给予不同浓度的褪黑素,能够减弱LPS诱导的心肌肥厚,降低心肌细胞TNF-a的分泌和TNFR1,Ca N和Ca MKⅡδB的表达和Ca2+内流,并且在浓度(1.5mg/L、3mg/L、6mg/L)范围内呈剂量依赖性。结论 在体外试验中褪黑素通过下调TNF-α和TNFR1的表达和维持Ca2+内稳态拮抗LPS诱导的心肌肥厚,发挥保护作用。
[Abstract]:Objective to study the protective effect and mechanism of melatonin on myocardial hypertrophy induced by lipopolysaccharide (LPS). Methods (1) the cultured cardiomyocytes were divided into five groups: control group, LPS melatonin (1.5mg/L) group, LPS melatonin (3mg/L) group, LPS melatonin (6mg/L) group. The morphological changes of cardiomyocytes were observed under inverted phase contrast microscope. (2) the protein level of cardiomyocytes in each group was detected by RC DCTM Protein Assay Kit. (3) the secretion of tumor necrosis factor 伪 (TNF- 伪) in cardiomyocytes was detected by enzyme-linked immunosorbent assay (ELISA). (4) the expression of tumor necrosis factor receptor 1 (TNFR1) gene and protein in cardiomyocytes of each group were detected by Realtime PCR and Western blot. (5) the Ca2 influx of cardiomyocytes in each group was recorded by Ca2 probe Fura-2/AM under imaging system. (6) the levels of CaN and CaMK 鈪，

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