胰岛素抵抗的慢性丙型肝炎患者外周血及肝组织内源性大麻素系统的变化

The change of endocannabinoid system in the peripheral circulation and liver in patients with chronic hepatitis C and insulin resistance

目的研究胰岛素抵抗(IR)的慢性丙型肝炎(CHC)患者外周血及肝组织内源性大麻素系统的变化。方法采用高效液相色谱-质谱联用技术检测CHC患者和CHC合并IR患者外周血内源性大麻素花生四烯酸乙醇胺(AEA)和花生四烯酰甘油(2-AG)水平,应用RT-PCR及Western印迹检测肝组织大麻素受体1(CB1R)及脂肪酸酰胺水解酶(FAAH)的表达,数据行t检验。结果与CHC患者相比,CHC合并IR患者外周血AEA水平升高[(6.80±1.70)pmol/mL比(3.88±0.63)pmol/mL,t=8.053,P<0.01];肝组织CB1R mRNA水平(1.7±0.4比1.0±0.2,t=4.696,P<0.01)和CB1R蛋白水平(0.8±0.2比0.4±0.1,t=5.367,P<0.01)增加;FAAH mRNA水平(0.5±0.1比1.0±0.3,t=4.743,P<0.01)和FAAH蛋白水平(0.5±0.1比0.8±0.3,t=2.846,P<0.01)下降。与胰岛素抵抗指数(HOMA-IR)<4的患者相比,HOMA-IR指数≥4的患者AEA水平升高[(7.53±1.85)pmol/mL比(5.56±1.40)pmol/mL(t=2.986,P<0.05)];肝组织CB1 mRNA水平(2.3±0.65比1.4±0.35,t=2.986,P<0.05)和CB1R蛋白水平(1.2±0.3比0.9±0.2,t=2.038,P<0.05)升高,FAAH mRNA水平(0.3±0.08比0.6±0.15,t=4.076,P<0.01)和FAAH蛋白水平(0.2±0.05比0.4±0.1,t=3.464,P<0.05)下降。结论随着IR程度加重,CHC患者外周血AEA水平升高;肝组织CB1R表达增加而FAAH表达减少。

Objective To study the change of endocannabinoid system in the peripheral circulation and liver in patients with chronic hepatitis C （CHC） and insulin resistance （IR） .Methods Circulating endocannabinoid （anandamide and 2-AG） levels were determined by liquid chromatography/mass spectrometry . The expression levels of cannabinoid receptor 1 （CB1R） and fatty acid amide hydrolase （FAAH） in liver was detected by RT-PCR and western blot .The data was checked by t test .Results Compared to patients with CHC ,in the patients with CHC and IR had increased ,the anandamide level was significantly increased in the peripheral circulation [（6 .80 ± 1 .70） vs （3 .88 ± 0 .63） pmol/ml ,t=8 .053 ,P〈0 .01] in patients with CHC and IR ,CB1R mRNA （1 .7 ± 0 .4 vs 1 .0 ± 0 .2 ,t=4 .696 ,P〈0 .01） and protein levels （0 .8 ± 0 .2 vs 0 .4 ± 0 .1 ,t=5 .367 ,P〈0 .01） in liver were significantly increased in patients with CHC and IR ,and FAAH mRNA level （0 .5 ± 0 .1 vs 1 .0 ± 0 .3 ,t=4 .743 ,P〈0 .01） and protein levels （0 .5 ± 0 .1 vs 0 .8 ± 0 .3 ,t=2 .846 , P〈 0 .01） were significantly decreased in patients with CHC and IR .Compared to patients with homeostasis model assessment of insulin resistance （HOMA-IR） 〈 4 ,in patients with HOMA-IR≥ 4 ,had increased the anandamide level increased[（7.53±1.85）vs（5.56±1.40）pmol/ml,t= 2.986,P〈 0.05）]wassignificantlyincreasedinpatientswith HOMA-IR≥4 ,CB1R mRNA level （2 .3 ± 0 .65 vs 1 .4 ± 0 .35 ,t=2 .986 ,P〈0 .05） and protein levels （1 .2 ± 0 .3 vs 0 .9 ±0 .2 ,t=2 .038 ,P〈0 .05） was significantly decreased in patients with HOMA-IR≥4 ,FAAH mRNA level （0 .3 ± 0 .08 vs 0 .6 ± 0 .15 ,t=4 .076 , P〈 0 .01） and protein levels （0 .2 ± 0 .05 vs 0 .4 ± 0 .1 , t= 3 .464 , P〈 0 .05） was significantly decreased in patients with HOMA-IR≥4 .Conclusion In patients with CHC and the development of IR ,the anandamide and CB1R levels were increasesd ,but level of FAAH was decreasesd in liver .