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补阳还五汤对动脉粥样硬化模型大鼠主动脉Rho激酶和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].

作者信息

Zhang Hong-zhen, Li Li, Jiao Rui, Zhang Ying, Qian Yan

出版信息

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2015 Dec;35(12):1495-500.

Abstract

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.

摘要

目的

观察益气活血法代表方剂补阳还五汤(BYHWD)对动脉粥样硬化(AS)模型大鼠主动脉中Rho相关卷曲螺旋形成蛋白丝氨酸/苏氨酸激酶(罗激酶,ROCK)及核转录因子κB(NF-κB)p65 mRNA表达和血脂水平的影响。

方法

采用维生素D3联合高脂饮食制备AS大鼠模型。将60只大鼠随机分为6组,即正常对照组、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)、辛伐他汀对照组(0.6 mg/kg)、补阳还五汤预防组(10 g/kg),每组10只。造模成功后所有药物干预28天。采用ELISA法检测氧化型低密度脂蛋白(ox-LDL)表达水平。采用酶法测定甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)水平。光镜下观察主动脉组织病理变化。采用实时荧光定量聚合酶链反应(RT-PCR)检测主动脉中Rho激酶和NF-κB p65的mRNA表达。

结果

高脂饮食联合腹腔注射维生素D3可诱导AS大鼠模型。AS模型大鼠主动脉形成典型动脉粥样硬化斑块。与正常对照组比较,模型组TC、TG、LDL-C及ox-LDL水平显著升高,HDL-C水平降低(P<0.01)。与模型组比较,补阳还五汤低、高剂量组、辛伐他汀对照组及补阳还五汤预防组TC、TG、LDL-C及ox-LDL水平均降低,HDL-C水平升高(P<0.05,P<0.01)。与补阳还五汤低剂量组比较,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上述指标降低更明显(P<0.05)。与正常对照组比较,模型组Rho激酶和NF-κB p65的mRNA表达水平显著升高(P<0.01)。与模型组比较,补阳还五汤低、高剂量组、辛伐他汀对照组及补阳还五汤预防组Rho激酶和NF-κB p65的mRNA表达明显降低(P<0.01)。与补阳还五汤低剂量组比较,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上述两个指标降低更明显(P<0.05)。但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血脂水平、Rho激酶或NF-κB p65的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Rho激酶和NF-κB p65的mRNA表达水平,降低血脂水平,对抗AS。抑制Rho激酶通路可能是其作用机制之一。

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