槲皮素对非酒精性脂肪性肝炎大鼠的作用及其机制研究

Effect of the quercetin on the non-alcoholic steatohepatitis in rats and its mechanism

目的探讨槲皮素对非酒精性脂肪性肝炎(NASH)模型大鼠的干预作用及对大鼠肝组织中脂联素、脂联素受体的影响。方法按照体重将雄性SD大鼠随机分为4组:正常组、模型组和低、高2个剂量实验组,每组9只。正常组予普通饲料,其余各组予高脂饲料10周建模。造模第2周,低、高2个剂量实验组灌胃40,80 mg·kg^-1·d^-1槲皮素(用1%羧甲基纤维素钠助溶);正常组和模型组灌胃等体积1%羧甲基纤维素钠。以化学发光法检测各组大鼠谷丙转氨酶(GPT)、谷草转氨酶(GOT)、碱性磷酸酶(ALP)、谷氨酰转肽酶(GGT)、三酰甘油(TG)和总胆固醇(TC)水平,以免疫印迹法检测各组大鼠肝内脂联素(ADPN)、脂联素受体2(Adipo R2)蛋白表达水平,以荧光定量PCR检测各组大鼠肝内ADPN、Adipo R2 m RNA表达水平。结果正常组、模型组和低、高2个剂量实验组的GPT分别为(43.51±1.52),(81.60±3.49),(69.15±2.78)和(53.98±2.01)U·L^-1;上述4组的GOT分别为(216.46±11.79),(302.30±29.34),(225.32±11.28)和(215.07±15.69)U·L^-1;上述这4组的ALP分别为(88.58±3.92),(176.63±11.72),(138.88±8.34)和(119.17±17.36)U·L^-1;上述这4组的GGT分别为(0.42±0.09),(1.43±0.17),(1.12±0.17)和(0.88±0.12)U·L^-1;上述这4组的TG分别为(0.16±0.02),(0.47±0.09),(0.43±0.08)和(0.23±0.06)mmol·L^-1;上述这4组的TC分别为(0.93±0.04),(1.72±0.09),(1.60±0.18)和(1.45±0.15)mmol·L^-1。上述指标:模型组与正常组相比、或者高剂量实验组与模型组相比,差异均有统计学意义(均P<0.05)。正常组、模型组和低、高2个剂量实验组的ADPN蛋白表达水平(OD值)分别为0.73±0.05,0.24±0.01,0.33±0.14和0.58±0.07;上述这4组的Adipo R2蛋白表达水平(OD值)分别为0.91±0.06,0.30±0.02,0.45±0.11和0.88±0.15;高剂量实验组与模型组比较,肝组织ADPN和Adipo R2蛋白水平均明显升高,差异均有统计学意义(均P<0.05);基因结果的趋势与蛋白一致。结论槲皮素能通过上调NASH模型大鼠ADPN、Adipo R2表达,有效改善肝功能、血脂,改善NASH大鼠肝组织脂肪变程度,减轻肝炎症。

Objective To investigate the effects of quercetin on nonalcoholic steatohepatitis(NASH)model rats and the effects of quercetin on adiponectin and adiponectin receptors on rat liver tissue.Methods The male Sprague-Dawley rats were randomly divided into 4 groups:normal group,model group,and experimental-L,experimental-H groups,with 9 rats in each group.The normal group was given normal feed,and the other groups were modeled for high fat feed for a total of 10 weeks.In the second week of moulding,quercetin 40,80 mg·kg^-1·d^-1 was administered intragastrically in low and high dose experimental groups(solubilization with 1%sodium carboxymethyl cellulos);1%sodium carboxymethyl cellulos was administered intragastrically in normal group,model group.Glutamic-pyruvic transaminase(GPT),glutamic-oxaloacetic transaminase(GOT),alkaline phosphatase(ALP),glutamyl transpeptidase(GGT),triglyceride(TG)and cholesterol(TC)were detected by chemiluminescence method.Western-blot was used to detect the protein expression levels of Adiponectin(ADPN)and Adiponectin receptor 2(AdipoR2)in the liver of each group.ADPN,AdipoR2 m RNA expression level in rats was was detected by fluorescence quantitative real-time PCR.Results The GPT in normal group,model group,experimental-L group,experimental-H group respectively were(43.51±1.52),(81.60±3.49),(69.15±2.78)and(53.98±2.01)U·L^-1;GOT in the above 4 groups respectively were(216.46±11.79),(302.30±29.34),(225.32±11.28)and(215.07±15.69)U·L^-1;ALP in the above 4 groups respectively were(88.58±3.92),(176.63±11.72),(138.88±8.34)and(119.17±17.36)U·L^-1;GGT in the above 4 groups respectively were(0.42±0.09),(1.43±0.17),(1.12±0.17)and(0.88±0.12)U·L^-1;TG in the above 4 groups respectively were(0.16±0.02),(0.47±0.09),(0.43±0.08)and(0.23±0.06)mmol·L^-1;TC in the above 4 groups respectively were(0.93±0.04),(1.72±0.09),(1.60±0.18)and(1.45±0.15)mmol·L^-1;comparing between model group and normal group,or experimental-H group and model group,the difference of the factors were significant(all P<0.05).The protein expression(OD value)of ADPN in normal group,model group,experimental-L group,experimental-H group respectively were 0.73±0.05,0.24±0.01,0.33±0.14 and 0.58±0.07;and protein expression(OD value)of AdipoR2 in the above 4 groups respectively were 0.91±0.06,0.30±0.02,0.45±0.11 and 0.88±0.15;comparing between experimental-H group and model group,the levels of ADPN and AdipoR2 protein in the liver tissue of experimental-H group were increased,the difference of the factors were significant(all P<0.05).The trend of gene results is consistent with that of protein.Conclusion Quercetin can up-regulate the expression of ADPN and AdipoR2 in NASH model rats,effectively improve liver function and blood lipids,improve the degree of hepatic steatosis in NASH rats,and reduce liver inflammation.