补阳还五汤对动脉粥样硬化模型大鼠主动脉Rho激酶及NF-κB p65 mRNA表达的影响

Effect of Buyang Huanwu Decoction on mRNA Expressions of Aorta Rho Kinase and NF-κB p65 in Atherosclerosis Model Rats

目的观察益气活血法代表方剂补阳还五汤对动脉粥样硬化(atherosclerosis,AS)模型大鼠主动脉组织Rho激酶及核转录因子-κB(nuclear transcription factor kappa B,NF-κB)p65 mRNA表达及血脂指标的影响。方法采用维生素D3联合高脂饮食法制备大鼠AS模型。将60只大鼠随机分为正常对照组、模型组、补阳还五汤低剂量组(10 g/kg)、补阳还五汤高剂量组(20 g/kg)、辛伐他汀组(0.6 mg/kg)及补阳还五汤预防组(10 g/kg)。造模成功后给药干预28天,采用ELISA法检测氧化型低密度脂蛋白(ox-LDL)水平,酶法测定血清TG、TC、低密度脂蛋白胆固醇(LDL-C)及高密度脂蛋白胆固醇(HDL-C)水平。光镜下观察主动脉组织AS病理变化。采用实时荧光定量PCR法检测主动脉Rho激酶及NF-κB p65 mRNA表达水平。结果高脂饮食加维生素D3腹腔注射可诱导大鼠形成AS模型,模型组大鼠主动脉有典型的粥样斑块形成。与正常对照组比较,模型组TC、TG及LDL-C及ox-LDL水平明显升高,HDL-C下降(P<0.01);与模型组比较,补阳还五汤高、低剂量组、辛伐他汀组及补阳还五汤预防组TC、TG、LDL-C及ox-LDL水平均下降,HDL-C升高(P<0.05,P<0.01);补阳还五汤高剂量组、辛伐他汀组及补阳还五汤预防组上述指标均较补阳还五汤低剂量组改善更明显(P<0.05)。与正常对照组比较,模型组Rho激酶及NF-κB p65 mRNA表达水平明显升高(P<0.01);与模型组比较,补阳还五汤高、低剂量组、辛伐他汀组及补阳还五汤预防组Rho激酶及NF-κB p65 mRNA表达水平明显降低(P<0.01);补阳还五汤高剂量组、辛伐他汀对照组及补阳还五汤预防组两项指标均较补阳还五汤低剂量组降低更明显(P<0.05),但3组间血脂、Rho激酶及NF-κB p65 mRNA表达水平比较,差异无统计学意义(P>0.05)。结论补阳还五汤可下调主动脉Rho激酶及NF-κB p65mRNA表达水平,降低血脂,具有抗AS作用,抑制Rho激酶通路可能是其作用机制之一。

Objective To observe the effect of Buyang Huanwu Decoction（ BYHWD）,a representative formula of qi benefiting blood activating method on aorta Rho associated coiled-coil forming protein serine / threonine kinase（ Rhokinase,ROCK） and nuclear transcription factor kappa B（ NF-κB） p65 mRNA expressions and levels of blood lipids in atherosclerosis（ AS） model rats. Methods The AS rat model was prepared by vitamin D3 and high fat diet. Totally 60 rats were randomly divided into 6 groups,i. e.,the normal control group,the model group,the low dose BYHWD group（ 10 g / kg）,the high dose BYHWD group（ 20 g / kg）,the Simvastatin control group（ 0. 6 mg / kg）,and the BYHWD prevention group（ 10 g/kg）,10 in each group. After successful modeling all medication was intervened for 28 days. Expression levels oxidized low density lipoprotein（ ox-LDL） were detected by ELISA. Levels of TG,TC,LDL-C,HDL-C were determined by enzyme method. Pathological changes of aortic tissue were observed under light microscope. mRNA expressions of Rho kinase and NF-κB p65 in aorta were detected by real time（ RT） PCR. Results High fat diet and peritoneal injection of vitamin D3 could induce AS rat model. Typical atheromatous plaque formed in aorta of AS model rats. Compared with the normal control group,levels of TC,TG,LDL-C,and ox-LDL significantly increased in the model group,but the HDL-C level decreased（P〈0. 01）. Compared with the model group,levels of TC,TG,LDL-C,and ox-LDL all decreased,but HDL-C increased in low and high dose BYHWD groups,the Simvastatin control group,and the BYHWD prevention group（P〈0. 05,P〈0. 01）. Compared with the low dose BYHWD group,above-mentioned indices were more obviously lowered in the high dose BYHWD group,the Simvastatin control group,and the BYHWD prevention group（P〈0. 05）. Compared with the normal control group,mRNA expression levels of Rho kinase and NF-κB p65 significantly increased in the model group（P〈0. 01）. Compared with the model group,mRNA expressions of Rho kinase and NF-κB p65 obviously decreased in low and high dose BYHWD groups,the Simvastatin control group,and the BYHWD prevention group（P〈0. 01）. Compared with the low dose BYHWD group,the two indicators were more obviously lowered in the high dose BYHWD group,the Simvastatin control group,and the BYHWD prevention group（P〈0. 05）. But there was no statistical difference in blood lipids levels,mRNA expression levels of Rho kinase or NF-κB p65 among the high dose BYHWD group,the Simvastatin control group,and the BYHWD prevention group（P〈0. 05）. Conclusions BYHWD could down-regulate mRNA expression levels of Rho kinase and NF-κB p65,lower levels of blood lipids,and fight against AS. Suppressing Rho kinase pathway might be one of its mechanisms.