虎杖苷调控JAK-STAT3信号通路对心肌缺血再灌注损伤的保护作用及机制

Protective effect and mechanism of polydatin on myocardial ischemia-reperfusion injury by regulating JAK-STAT3 signal

目的研究虎杖苷对氧糖剥夺/再复氧(oxygen-glucosed/deprivation,OGD/R)条件下对心肌细胞的保护作用及机制研究。方法H9C2 细胞在常氧和OGD/R 条件下培养,采用20、40、80 μmol/L 虎杖苷和渥曼青霉素(Wortmannin)处理。细胞计数试剂盒(cell counting kit,CCK)-8检测H9C2细胞在不同条件下的存活能力,酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测培养细胞上清肿瘤坏死因子(tumornecrosis factor,TNF-α)和白细胞介素6(interleukin-6,IL-6)的浓度,蛋白免疫印迹(Western Blot)检测总STAT3、JAK、磷酸化STAT3和磷酸化JAK 的蛋白浓度。结果(1)虎杖苷明显改善了OGD/R 条件下细胞的存活能力,并且抑制了OGD/R 条件下诱导产生的TNF-α和IL-6 的分泌;(2)虎杖苷诱导H9C2 细胞JAK-STAT3信号通路;(3)抑制JAK-STAT3信号通路会抵消虎杖苷抗炎和促进细胞生存能力的作用。结论OGD/R条件下,虎杖苷提高了H9C2 细胞的存活率,同时其可以通过磷酸化JAK 和STAT3 蛋白表达激活JAK-STAT3 信号通路,从而对心肌缺血再灌注损伤起到保护作用。

Objectives To study the protective effect and mechanism of polydatin on myocardial cells under oxygen glucose deprivation/reoxygenation(OGD/R)conditions. Methods H9C2 cells were cultured under normoxia and OGD/R conditions and treated with 20,40,80 μmol/L of polydatin and wortmannin. Cell counting kit(CCK)-8 was used to detect the survival ability of H9C2 cells under different conditions. The concentration of tumor necrosis factor(TNF)-α and interleukin-6(IL-6)were detected by enzyme-linked immunosorbent assay(ELISA). The total STAT3,JAK and phosphorylation of JAK and STAT3 protein expression were detected by Western Blot(WB). Results (1)Polydatin significantly improved the viability of cells under OGD/R conditions and inhibited the secretion of TNF-α and IL-6 induced by OGD/R conditions.(2)JAK-STAT3 signaling path way was induced by polydatin.(3)Inhibition of JAKSTAT3 signaling pathway by Hooks in the H9C2 cells counteracted the anti-inflammatory and cell viability of polydatin. Conclusions Under OGD/R conditions,polydatin increases the survival rate of H9C2 cells,and can activate JAKSTAT3 signaling through phosphorylation of JAK and STAT3 protein expression,thus protecting myocardial ischemia/ reperfusion injury.