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目的探讨乙酰辅酶A羧化酶2(ACC2)表达下调对高糖培养的人肾小管上皮细胞(HKC)脂质沉积及小管上皮细胞间充质转化(EMT)的影响及可能机制。方法构建ACC2基因特异性的短发夹双链RNA(shRNA)慢病毒感染HKC,将其分为5组:正常糖组(5.5 mmol/L葡萄糖,NG组)、高糖组(30 mmol/L葡萄糖,HG组)、高渗对照组(5 mmol/L葡萄糖+25 mmol/L甘露醇,HO组)、ACC2干扰组(感染含ACC2干扰序列慢病毒,ACC2-shRNA组)、干扰对照组(感染携带绿色萤光蛋白的阴性对照慢病毒,NC-shRNA组),分别处理96 h,Western blotting检测ACC2干扰效率及E-钙黏蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)表达,油红O染色检测细胞内脂质沉积,光镜下观察细胞形态变化。结果与HO组相比,HG组HKC细胞内脂质沉积增加,细胞拉长,E-cadherin表达减少,α-SMA表达增多(P<0.05);干扰ACC2基因后,高糖诱导的细胞内脂质沉积减少,细胞形态改善,E-cadherin和α-SMA表达趋于正常。结论下调ACC2表达可抑制高糖诱导的HKC脂质沉积和小管上皮细胞EMT。
Objective To investigate the effects of down-regulation of acetyl-CoA carboxylase 2 (ACC2) on lipid deposition and tubular epithelial cell mesenchymal transition (EMT) in cultured human renal tubular epithelial cells (HKC) and its possible mechanism. Methods ACC2 gene-specific short hairpin RNA (shRNA) lentivirus was transfected into HKC and divided into 5 groups: normal glucose group (5.5 mmol / L glucose, NG group), high glucose group (30 mmol / L HG group), hypertonic control group (5 mmol / L glucose + 25 mmol / L mannitol, HO group), ACC2 interference group (ACC2-shRNA group infected with ACC2 interference RNA interference group) Infected with green fluorescent protein-negative control lentivirus, NC-shRNA group) for 96 h respectively. Western blotting was used to detect the ACC2 interference efficiency and the expression of E-cadherin, α-smooth muscle actin (α- SMA) expression, lipid red O staining intracellular lipid deposition, morphological changes observed under light microscope. Results Compared with HO group, the deposition of lipid in HKC cells of HG group was prolonged, the expression of E-cadherin and the expression of α-SMA were increased in the HG group (P <0.05). After interference with ACC2 gene, the intracellular lipid Deposition decreased, cell morphology improved, E-cadherin and α-SMA expression tended to be normal. Conclusion Down-regulation of ACC2 expression can inhibit high glucose-induced HKC lipid deposition and tubular epithelial cell EMT.