大鼠肾周脂肪组织趋化因子样受体1过表达对非酒精性脂肪性肝炎的影响

Role of CMKLR1 over-expression in adipose tissues in rat with nonalcoholic steatohepatitis

目的探讨慢病毒介导的趋化因子样受体1(CMKLR1)在大鼠肾周脂肪组织中过表达对非酒精性脂肪性肝炎的影响。方法将42只雄性SD大鼠随机分为对照组、模型组、转染组各12只和空转染组6只。给予对照组普通饮食喂养,给予其余组高脂饮食喂养。在造模开始时给予各组经尾静脉注射溶媒和真病毒或空病毒。在实验第8 w和12 w,分批处死大鼠,取肝组织行苏木精-伊红染色,观察组织病理学变化;采用酶联免疫吸附法测定血清凯莫瑞和脂联素水平;分别采用RT-PCR法和Western blot法测定肾周脂肪组织CMKLR1和脂联素mRNA和它们的蛋白表达。结果模型组大鼠肝组织炎症程度明显于对照组和转染组;在实验8 w和12 w,模型组血清凯莫瑞水平分别为(4.65±0.48)mmol/L和(4.47±0.37)mmol/L,与对照组水平无明显差异[(4.38±0.43)mmol/L和(4.16±0.27)mmol/L],但均显著低于转染组水平[(7.66±0.53)mmol/L和(6.74±0.59)mmol/L,P<0.05];在8 w时,模型组脂肪组织CMKLR1 mRNA及其蛋白相对量分别为(0.235±0.008)和(0.206±0.005),与对照组水平无显著差异[(0.226±0.008)和(0.21±0.117)],但显著低于转染组水平[(0.579±0.016)和(0.504±0.121),P<0.05];第12 w时,脂肪组织CMKLR1 mRNA及其蛋白表达水平与第8 w时结果相似;在8 w时,模型组大鼠血清脂联素水平、脂肪组织脂联素mRNA及其蛋白表达水平分别为[【(31.82±1.51)mmol/L、(0.126±0.005)和(0.14±0.008)],显著低于对照组[(37.92±4.09)mmol/L、(0.262±0.007)和(0.35±0.016),P<0.05]和转染组水平[(36.50±2.43)mmol/L、(0.193±0.058)和(0.232±0.012),P<0.05];在12 w时,所测血清脂联素水平、脂肪组织脂联素mRNA水平及其蛋白表达水平与第8 w时结果相似。结论慢病毒介导的大鼠脂肪组织内过表达CMKLR1可显著改善大鼠非酒精性脂肪性肝炎时肝组织病理学损害。

Objective To study the role of ehemokine-like receptor 1 （CMKLR1） over-expression evoked by lentivirus in the adipose tissues of rats with nonalcoholic steatohepatitis （NASH）. Method 42 male SD rats were randomly divided into four groups. Rats in control group （n=6） fed with normal diet,and rats in other three fed with high fat diet. Vehicle,CMKLR1 virus （2×10^9pfu） or control virus （2×10^9pfu） was injected via tail vein at the first day of experiment. Rats were sacrificed at the end of 8 and 12 weeks. The pathological changes of liver tissues were observed. Kay Murray and adiponeetin in serum were measured, and the levels of adiponectin in adipose tissues and the mRNA and its protein levels of CMKLR1 in adipose tissues were detected by RT-PCR or Western blot. Results The pathological inflammation in liver tissues of rats with CMKLR1 overexpression was significantly improved as compared with that in the controls or rats with control virns;The serum levels of ehemerin at 8 and 12 weeks were（4.65±0.48） mmol/L and（4.47±0.37）mmol/L）,respectively,similar to those in the controls [（4.38±0.43） mmol/L and （4.16±0.27）mmol/L],and were significantly lower than those in rats with CMKLR1 overexpression [（7.66±0.53）mrnol/L and （6.74±0.59）mnmol/L,respeetively,P〈0.05];CMKLR1 mRNA and its protein in model group at 8 weeks were （0.235±0.008） and （0.206±0.005）,respectively,similar to those in the controls [（0.226±0.008） and （0.21±0.117）],and they were significantly lower than those in rats with CMKLR1 overexpression [（0.579±0.016） and （0.504±0.121）,respectively,P〈0.05];A similar changes in CMKLR1 mRNA and its protein were observed at 12 weeks;At 8 weeks,serum adiponectin,adiponectin mRNA and its protein in model group were [（31.82±1.51）mmol/L, （0.126±0.005） and （0.14±0.008）],which were significantly lower than those in the controls at the same period [ （37.92±4.09）mmol/L, （0.262±0.007） and（0.35±0.016）, P〈0.05 ], and those in the transfection group [（36.50±2.43）mmol/L, （0.193±0.058） and （0.232±0.012）,P〈0.05];A similar changes in serum adiponectin,adiponectin mRNA and its protein were observed at 12 weeks. Conclusion CMKLRI over- expression evoked by lentivirus transfection could improve pathological inflammation in rats with NASH.