外源性硫化氢对高糖诱导的肾小管上皮细胞凋亡的影响

Influence of exogenous hydrogen sulfide on cell apoptosis induced by high glucose in human proximal renal tubular epithelial cells

目的观察硫氢化钠(NaHS,外源性的H2S供体)对低糖和高糖环境下的人肾小管上皮细胞(HK-2)凋亡的影响。方法传代培养人近曲肾小管上皮细胞,细胞用无血清培养基同步化24 h后,将细胞分为8组:低糖对照组(5.5 mmol/L D-葡萄糖、低糖+不同剂量NaHS组(50、100、200μmol/L)、高糖组(40 mmol/L D-葡萄糖)、高糖+不同剂量NaHS组(50、100、200μmol/L)。将细胞置于37℃,5%CO_2培养箱中刺激培养24、48 h,收集细胞。用流式细胞仪测定细胞凋亡率。结果 (1)刺激培养24、48 h,低糖+不同剂量NaHS组HK-2细胞凋亡率明显低于低糖对照组(P<0.05),以NaHS中剂量组降低明显。(2)高糖刺激培养24、48 h,高糖组细胞HK-2细胞凋亡率明显高于低糖对照组(P<0.01)。(3)高糖+不同剂量NaHS组HK-2细胞凋亡率明显低于高糖组(P<0.05),以NaHS中剂量组降低明显。结论 NaHS能抑制低糖条件下肾小管上皮细胞的凋亡以及高糖诱导的肾小管上皮细胞的凋亡,有助于糖尿病肾病的治疗。

Objective To investigate the influence of sodium hydrosulfide （NariS, was taken as a donor of H2 S） on cell apoptosis induced by high glucose in human proximal renal tubular epithelial cells （HK -2 ）. Methods Human renal proximal tubular cells were cultured and divided into eight groups：low glucose as control group （5.5 mmol/L D -Glucose） ,low glucose ＋ NariS pretreatment group with dose of 50,100 and 200 txmol/L,high glu- cose group（40 mmol/L D -Glucose） ,high glucose ＋ Naris pretreatment group with dose of 50,100 and 200 μmol/L. Each group has three samples,cultured in 37 ℃ and 5% CO2incubator for 24 and 48 h, collected cells and cell supernatant fluid. Apoptosis rate was measured by flow cytometry. Results When cells were cultured 24 and 48 h, the apoptosis rate of low glucose ＋ NariS pretreatment groups were significantly lower than low glucose group （P 〈 0.05） ,and decreased significantly in low glucose ＋ NariS （100txmol/L） dose group. The apoptosis rate of high glucose group was significantly higher than low glucose group （ P 〈 0.05 ）, high glucose ＋ NariS pre- treatment groups were significantly lower than high glucose group （P 〈 0.05 ） , and decreased significantly in high glucose ＋ NariS（ 100 txmol/L）dose group. Conclusion NariS can suppress cell apoptosis induced by low glu- cose and high glucose in human proximal renal tubular epithelial cells, which could be a novel approach of trea- ting diabetic nephropathy.