Vasostatin-1对缺氧复氧肾小管上皮细胞炎性损伤的影响

Effect of vasostatin-1 on inflammatory damage of hypoxic reoxygenation in renal tubular epithelial cells

目的探讨血管生成抑制因子vasostatin-1(VS-1)对缺氧复氧肾小管上皮细胞的炎性损伤作用及机制。方法采用缺氧复氧方式制造肾小管上皮细胞炎性损伤。构建慢病毒载体并以脂质体法将shVS-1转染NRK-52E细胞。ELISA法检测细胞中LDH和IL-1β含量;Western blot检测细胞中VS-1、pro-caspase-1和caspase-1蛋白表达。结果与对照组相比,缺氧复氧组细胞LDH和IL-1β含量显著升高,且VS-1蛋白表达显著升高。成功构建沉默VS-1的慢病毒载体,且转染缺氧复氧NRK-52E细胞,可降低细胞中LDH和IL-1β含量、降低caspase-1蛋白表达,过表达VS-1则具有相反的功能。结论沉默VS-1可保护缺氧复氧肾小管上皮细胞的炎性损伤,可能与下调caspase-1有关,可为急性肾损伤的临床治疗提供依据。

Objective To study the inflammatory injury mechanism of vasostatin-1(VS-1)in renal tubular epithelial cells by hypoxic reoxygenation.Methods Inflammatory injury of kidney tubular epithelial cells was established by hypoxia reoxygenation.The lentiviral vector of shVS-1 was transfected into NRK-52E cells by liposome method.LDH and IL-1βlevels in cells were detected by ELISA.The protein expression of VS-1,pro-caspase-1 and caspase-1 in cells were detected by Western blot.Results Compared with the control group,the levels of LDH and IL-1βin the hypoxia reoxygenation group were significantly increased,and the protein expression of VS-1 was significantly increased.The lentiviral vector was successfully constructed.Transfection of silencing VS-1 lentiviral vector into NRK-52E cells with hypoxic reoxygenation could decrease LDH and IL-1βlevels and downregulate caspase-1 expression,but overexpression of VS-1 possessed the opposite function.Conclusion Silencing VS-1 could protect the kidney tubular epithelial cells with hypoxic reoxygenation from inflammatory injury,which may be related to the down-regulation of caspase-1,which will provide a basis for clinical treatment of acute kidney injury.