摘要
目的观察炎症状态下高脂介导的内质网应激在人肾脏系膜细胞(HMCs)损伤中的作用,并探讨高脂在肾脏损伤中的作用机制。方法体外培养HMCs,分为对照组、高脂组、炎症组、高脂+炎症组、高脂+炎症+4-苯基丁酸(4-PBA)干预组(4-PBA干预组),分别培养24、48和72h。用油红O染色观察HMCs胞内脂质沉积水平;四甲基偶氮唑盐(MTT)法检测细胞增殖变化;实时荧光定量PCR检测GRP78、纤连蛋白(FN)mRNA水平;免疫细胞化学半定量分析糖调节蛋白78(GRP78)蛋白水平。结果 (1)与对照组比较,高脂组、炎症组、高脂+炎症组HMCs胞内脂质沉积增多(P均<0.01),细胞增殖增快,且呈时间依赖性(P均<0.01),同时FN mRNA表达升高(P均<0.01);与高脂+炎症组比较,4-PBA干预组胞内脂质沉积减少(P均<0.01),细胞增殖减慢(P均<0.01),且FN mRNA表达降低(P<0.05,P<0.01)。(2)与对照组比较,高脂组、炎症组、高脂+炎症组GRP78mRNA和蛋白表达升高(P<0.05,P<0.01);而4-PBA干预组GRP78mR-NA和蛋白表达较高脂+炎症组降低(P均<0.01)。(3)在24、48及72h,细胞脂质沉积水平与其增殖、GRP78蛋白及FN mR-NA水平均呈正相关(P均<0.01),GRP78蛋白表达水平与细胞增殖和FN mRNA水平呈正相关(P均<0.05)。结论炎症状态下,高脂可通过启动HMCs内质网应激致细胞损伤。
Objective To study the role of high lipid-mediated endoplasmic reticulum stress in human glomerular mesangial cells （HMCs） injury under inflammation condition, and to explore the mechanism of lipid-induced kidney injury. Methods HMCs were divided into control group, high lipid group, inflammatory stress group, high lipid plus inflammatory stress group, and high lipid plus inflammatory stress plus 4-phenylbutyrie acid （4-PBA） group. After culture for 24 h, 48 h, and 72 h, Oil red O staining was used to evaluate lipid droplet accumulation in the cells; cell proliferation was assessed by MTT assay; expression of GRP78 protein was measured by immunocytochemistry method; and the levels of GRP78 and FN mRNA were examined by real-time PCR. Results （1） Compared with control group, the high lipid group, inflammatory stress group and high lipid plus inflammatory stress group had significantly increased intraeellular cholesterol levels （all P 〈 0. 01）, significantly higher FN mRNA levels （all P〈0.01）, and significantly faster cell proliferation （all P〈0.01）, which was in a time-dependent manner. Compared with high lipid plus inflammatory stress group, 4-PBA group had significantly reduced lipid deposition （all P〈0.01）, slower cell proliferation （all P〈0. 01） and lower FN mRNA expression （P〈0.05, P〈0. 01） induced by high lipid. （2） Compared with control group, high lipid group, inflammatory group and high lipid plus inflammatory stress group had significantly increased expression of GRP78 mRNA and protein （P〈0.05 ,P〈0.01）, and the expression in 4- PBA treatment group was significantly lower than that in the high lipid plus inflammatory stress group （all P〈0.01）. （3）After treatment with high lipid or inflammation for 24 h, 48 h and 72 h, the intracellular cholesterol level was positively correlated with mesangial cell proliferation and expression of GRP78 protein and FN mRNA （all P〈0.01）. The cellular expression o〈 GRP78 protein was positively cor
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2013年第4期375-381,共7页
Academic Journal of Second Military Medical University
基金
国家自然科学基金(81270802)
重庆市自然科学基金(CSTC
2009BA5080)~~
关键词
内质网应激
肾小球系膜细胞
高脂
炎症
endoplasmic reticulum stress
mesangial cells
high lipid
inflammation
