摘要
目的:探讨柚皮苷能否通过抑制瘦素通路保护H9c2心肌细胞,对抗高糖HG引起的损伤。方法:应用Westernblotting法检测瘦素及瘦素受体(LEPR)的表达水平;细胞计数盒(CCK-8)测定细胞存活率;Hoechst33258核染色荧光显微镜照相测定凋亡细胞的形态及数量的变化;双氯荧光素(DCFH-DA)染色荧光显微镜照相法检测胞内活性氧(ROS)水平;罗丹明123染色法测定线粒体膜电位(MMP)。结果:应用35mmol/L葡萄糖(HG)处理H9c2心肌细胞6~24h能上调瘦素表达,其中9h时表达最多;HG处理心肌细胞1-24h也能促进LEPR表达,其中12h时表达最多;在HG处理心肌细胞前,80μmol/L柚皮苷预处理2h能明显抑制HG对瘦素及LEPR表达的上调作用;柚皮苷预处理2h、瘦素拮抗剂预处理24h或瘦素受体拮抗剂预处理2h均能抑制HG引起的心肌细胞损伤,使细胞存活率增多,凋亡细胞数量和胞内ROS生成减少,MMP回升。结论:柚皮苷可通过抑制瘦素通路保护H9c2心肌细胞,对抗HG引起的损伤。
AIM: To study whether naringin protects H9c2 cardiac cells against high glucose (HG)-induced injury by inhibiting the leptin pathway. METHODS: The expression levels of leptin and leptin receptor (LEPR) were de- tected by Western blotting. The cell viability was analyzed by CCK-8 assay. The changes of the morphology and the number of apoptotic cells were tested by Hoeehst 33258 nuclear staining. The intracellular levels of reactive oxygen species (ROS) were measured by DCFH-DA staining. Mitoehondrial membrane potential (MMP) was determined by rhodamine 123 stai- ning. RESULTS: Treatment of the ceils with 35 mmol/L glucose (HG) for 6 ~ 24 h up-regulated the expression of leptin in H9e2 cardiac cells with the peak value at 9 h. Treatment of the cells with HG for 1 - 24 h also enhanced the expression of LEPR, peaking at 12 h. Pretreatment with 80 Ixmol/L naringin for 2 h before exposure of the H9c2 cardiac cells to HG significantly inhibited the up-regulation of both leptin and LEPR induced by HG. Pretreatment of the cells with naringin for 2 h, leptin antagonist for 24 h, or leptin receptor antagonist for 2 h attenuated HG-induced injury in the cardiomyocytes, evidenced by an increase in cell viability, decreases in the number of apoptotie cells and intracellular ROS production as well as a recovery of MMP. CONCLUSION: Naringin may protect the cardiomyocytes against the HG-induced injury by inhibition of the leptin pathway.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2014年第3期423-429,共7页
Chinese Journal of Pathophysiology
基金
湛江市财政资金科技专项竞争性分配项目(No.湛科[2011]79号)
广东省重大科技专项(No.2012A080202020)
关键词
柚皮苷
瘦素
受体
瘦素
高血糖
心肌细胞
Naringin
Leptin
Receptors, leptin
Hyperglycemia
Cardiomyocytes