氧化应激和P53参与波动高糖诱导的肾小管上皮细胞凋亡

目的: 探讨波动性高糖引起的肾小管上皮细胞凋亡的机制。方法: 体外培养人肾小管上皮细胞 (HK-2),给予稳定高糖或波动高糖干预,并使用抗氧化剂和P53特异性抑制剂验证氧化应激和P53在波动高糖诱导的肾小管上皮细胞凋亡中的作用;此外,SD大鼠随机分为正常对照组(A)、糖尿病稳定高血糖组(B)和糖尿病波动高血糖组(C)。采用链脲佐菌素(STZ)65 mg/kg腹腔注射诱发糖尿病,血糖波动组每天定时腹腔注射速效胰岛素,并错时给予葡萄糖,造成一天中血糖浓度大幅波动模型。采用比色法检测超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,免疫组化法和Western blotting检测NADPH氧化酶4(Nox4)和P53蛋白表达,流式细胞术和原位缺口末端标记法(TUNEL)检测肾脏细胞凋亡。结果: (1)与稳定高糖比较,波动高糖引起更加明显的HK-2细胞凋亡,P53蛋白表达明显上调,同时培养上清液中MDA含量增加和SOD活性下降,抗氧化剂和P53抑制剂可明显抑制磷酸化P53蛋白表达和细胞凋亡。(2)在体动物实验制模12周后,与B组比较,C组肾组织Nox4表达明显增加,肾小管磷酸化P53蛋白表达上调,细胞凋亡数增加。结论: 氧化应激和P53参与波动性高糖诱导的肾小管上皮细胞凋亡。

Oxidative stress and P53 involve in fluctuant high glucose-induced apoptosis of renal tubular epithelial cells

AIM: To explore the mechanism of fluctuant high blood glucose-induced apoptosis of renal tubular epithelial cells.METHODS: Cultured human renal tubular epithelial cells(HK-2) were treated with stable high glucose or fluctuant high glucose.Antioxidant and specific inhibitor of P53 were applied for identifying the role of oxidative stress and P53 in fluctuant high glucose-induced apoptosis of renal tubular epithelial cells.Additionally,SD rats were randomly divided into normal control group(A),stable high blo...