目的 基于CYP2J3/EETs介导脂肪酸代谢探讨化瘀祛痰方防治血脂异常的机制。方法 32只SPF级SD大鼠分为空白对照组、模型组、化瘀祛痰方组、辛伐他汀组，每组8只。除空白对照组外，其他各组给予高脂饲料喂饲，建立高脂血症模型。于第13周开始，化瘀祛痰方组给予化瘀祛痰方灌胃，辛伐他汀组给予辛伐他汀灌胃，空白对照组、模型组给予相应体积的生理盐水灌胃，每日1次，灌胃4周。采用全自动生化分析仪检测各组大鼠血清中总胆固醇(Total cho-lesterol，TC)、甘油三酯(Triglyceride，TG)、低密度脂蛋白胆固醇(Low density lipoprotein cholesterol，LDL-C)及高密度脂蛋白胆固醇(High density lipoprotein cholesterol，HDL-C)水平;HE染色观察肝脏组织病理变化;Elisa检测各组大鼠肝脏14，15-环氧二十碳三烯酸(14，15-epoxyeicosatrienoic acid，14，15-EET)水平;Real time RT-PCR 检测肝脏组织细胞色素 P450 表氧化酶 2J3(Cytochrome P450 oxidase 2J3，CYP2J3)、肉毒碱棕榈酰转移酶 1(Carnitine palmitoyltransferase-1，CPT-1)mRNA 水平;Western blot 法检测肝脏组织 CYP2J3、腺苷酸 活化蛋 白激酶(AMP-actived protein kinase，AMPK)、磷酸化腺苷酸活化蛋白激酶(p-AMPK)、乙酰辅酶A羧化酶(Acetyl CoA Carboxylase，ACC)、磷酸化乙酰辅酶A羧化酶(p-ACC)、CPT-1蛋白水平。结果 与空白对照组比较，模型组大鼠血清TC、TG和LDL-C水平显著升高(P＜0。01)，HDL-C水平显著降低(P＜0。05);肝脏脂质沉积显著，可见大量脂滴;肝脏组织内14，15-EET含量显著降低(P＜0。01);肝脏 CYP2J3、CPT-1 mRNA 和蛋白表达水平显著降低(P＜0。01)，p-AMPK/AMPK、p-ACC/ACC 显著降低(P＜0。01)。与模型组比较，经化瘀祛痰方及辛伐他汀干预后，大鼠血清TC、TG和LDL-C水平显著降低(P＜0。05或P＜0。01)，HDL-C水平显著升高(P＜0。05);肝脏脂质沉积情况明显改善，脂滴数明显减少;肝脏组织内14，15-EET水平显著升高(P＜0。01);肝脏CYP2J3、CPT-1 mRNA和蛋白表达水平显著升高(P＜0。05或P＜0。01)，p-AMPK/AMPK、p-ACC/ACC显著升高(P＜0。05或P＜0。01)。结论 化瘀祛痰方可有效改善高脂血症大鼠血脂异常，减轻肝脏脂质沉积情况，其机制可能与CYP2J3/EETs介导脂肪酸代谢密切相关。
Effects of Huayu Qutan Prescription(化瘀祛痰方)on Liver CYP2J3/EETs Mediated Fatty Acid Metabolism in Hyperlipidemia Rats
Objective To investigate the mechanism of Huayu Qutan Prescription(化瘀祛痰方)in the prevention and treat-ment of dyslipidemia based on cytochrome P450 2J3(CYP2J3)/EETs mediated fatty acid metabolism.Methods Thirty-two SD rats were divided into normal group,model group,Huayu Qutan Prescription group and simvastatin group,with 8 rats in each group.Except the normal group,the other groups were given high-fat feed to establish the hyperlipidemia model.From the 13th week,the Huayu Qutan Prescription group was given Huayu Qutan Prescription by gavage,the simvastatin group was given simv-astatin by gavage,and the normal group and the model group were given the corresponding volume of normal saline by gavage,once a day for 4 weeks.The serum levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C)were detected.Observation of pathological changes in liver tissue by HE staining.The levels of 14,15-epoxyeicosatrienoic acid(14,15-EET)in liver of rats in each group were detected by ELISA.The mRNA levels of CYP2J3 and carnitine palmitoyltransferase-1(CPT-1)in liver tissue were detected by RT-PCR,and the protein levels of CYP2J3,phosphorylation AMP-actived protein kinase(p-AMPK)/AMP-actived protein kinase(AMPK),phosphorylation acetyl CoA carboxylase(p-ACC)/acetyl CoA carboxylase(ACC)and CPT-1 in liver tissue were detected by Western blot.Results Compared with those of the normal group,the contents of TC,TG and LDL-C in the serum in the model group were significantly increased(P＜0.01),the HDL-C level was significantly decreased(P＜0.05),the lipid deposition was observed,the lipid droplets in the liver were significantly increased,the 14,15-EET content in the liver tissue was significantly decreased(P＜0.01),the mRNA expressions of CYP2J3 and CPT-1 in liver were significantly decreased(P＜0.01),the pro-tein expressions of CYP2J3 and CPT-1 were significantly decreased(P＜0.01),and the levels of p-AMPK/AMPK and p-ACC/ACC were significantly decreased(P＜0.01).Compared with those of the model group,the contents of TC,TG and LDL-C were significantly decreased(P＜0.05 or P＜0.01),the level of HDL-C was significantly increased(P＜0.05),the lipid deposition was significantly improved and the lipid droplets in the liver were significantly decreased by HE staining,the level of 14,15-EET in the liver was significantly increased after the intervention of Huayu Qutan Prescription and simvastatin(P＜0.01),the mRNA expressions of CYP2J3 and CPT-1 in liver were significantly increased(P＜0.05 or P＜0.01),the protein expressions of C YP2 J3 and CPT-1 were significantly increased(P＜0.05 orP＜0.01)and the levels of p-AMPK/AMPK and p-ACC/ACC were significantly increased(P＜0.05 or P＜0.01).Conclusion Huayu Qutan Prescription can effectively im-prove the dyslipidemia of hyperlipidemia rats and reduce the liver lipid deposition.Its mechanism may be closely related to the CYP2J3/EETs mediated fatty acid metabolism.
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