Netrin-1对高糖诱导的肾小管上皮细胞损伤的影响  

Effect of netrin-1 on high glucose induced injury of renal tubular epithelial cells

作　　者：赵静[1] 翟铁[2] 梁宏霞[1] ZHAO Jing;ZHAI Tie;LIANG Hong-xia(Department of Clinical Medicine,Tangshan Vocational and Technical College,Tangshan 063000,China;Department of Endocrinology,Chengde Central Hospital,Chengde 067000,China)

机构地区：[1]唐山职业技术学院临床医学系,河北唐山063000 [2]承德市中心医院内分泌科,河北承德067000

出　　处：《中国病理生理杂志》2019年第2期231-236,共6页Chinese Journal of Pathophysiology

基　　金：河北省医学科学研究重点课题(No.20160307)

摘　　要：目的:研究netrin-1对高糖诱导的肾小管上皮细胞损伤的影响。方法:用高糖培养人肾小管上皮HK-2细胞,real-time PCR和Western blot测定细胞中netrin-1的表达水平。用过表达netrin-1慢病毒感染HK-2细胞,检测其对高糖环境下HK-2细胞中netrin-1表达的影响。流式细胞术测定细胞凋亡,Western blot测定cleavedcaspase-3蛋白水平,2,4-二硝基苯肼法检测培养液中乳酸脱氢酶(LDH)活性,硫代巴比妥酸法检测培养液中丙二醛(MDA)含量,ELISA法测定培养液中白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)含量。结果:高糖处理后的HK-2细胞中netrin-1的mRNA和蛋白水平均明显下调(P <0.05)。过表达netrin-1慢病毒能够上调高糖环境下HK-2细胞中netrin-1的表达水平(P <0.05)。高糖可促进肾小管上皮细胞分泌IL-1β和TNF-α,提高细胞培养液中LDH和MDA的水平(P <0.05),使肾小管上皮细胞的caspase-3活化并诱导细胞凋亡;上调netrin-1的HK-2细胞经高糖诱导后,细胞分泌的IL-1β和TNF-α减少,细胞中活化的caspase-3蛋白水平降低,培养液中LDH和MDA水平降低,细胞凋亡减少(P <0.05)。结论:上调netrin-1减轻高糖诱导的肾小管上皮细胞氧化损伤和炎性损伤,减少细胞凋亡。AIM:To study the effect of netrin-1 on the damage of renal tubular epithelial cells induced by high glucose.METHODS:Human renal tubular epithelial HK-2 cells were treated with high glucose.Real-time PCR and Western blot were used to detect the expression level of netrin-1 in the cells.HK-2 cells were infected with netrin-1-over-expressing lentivirus,and the effect of netrin-1 over-expression on the HK-2 cells treated with high glucose was observed.The apoptosis rate was analyzed by flow cytometry.The protein level of cleaved caspase-3 was determined by Western blot.lactate dehydrogenase(LDH)activity in the culture medium was measured by 2,4-binitrobenzene hydrazine method.The content of malondialdehyde(MDA)in the culture medium was detected by thiobarbituric acid method.The concentrations of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the culture medium were measured by ELISA.RESULTS:The expression of netrin-1 at mRNA and protein levels in the HK-2 cells after high glucose treatment was significantly lower than that in the control cells(P<0.05).Infection with netrin-1-over-expressing lentivirus up-regulated the expression of netrin-1 in the HK-2 cells treated with high glucose.High glucose promoted the secretion of IL-1β and TNF-α,decreased the levels of LDH and MDA in the cell culture supernatant,and induced apoptosis and activation of caspase-3 in renal tubular epithelial cells(P<0.05).After the HK-2 cells with up-regulation of netrin-1 were induced by high glucose,the IL-1β and TNF-α secretion,the levels of LDH and MDA in the culture medium,the apoptosis,and the level of activated caspase-3 protein in the cells were all decreased,as compared with the control cells(P<0.05).CONCLUSION:Up-regulation of netrin-1 expression attenuates oxidative damage and inflammatory injury,and reduces apoptosis induced by high glucose in renal tubular epithelial cells.

关 键 词：肾小管上皮细胞 NETRIN-1 糖尿病肾病 氧化损伤 细胞凋亡 

分 类 号：R587.2[医药卫生—内分泌] R363.2[医药卫生—内科学]