颅内消瘀汤对大鼠颈动脉球囊损伤后血管内皮AMPK/eNOS信号的影响

Effect of Lunei Xiaoyu Decoction on Vascular Endothelial AMPK/eNOS Signal in Balloon Injury Model of Common Carotid Artery in Rats

目的观察颅内消瘀汤对大鼠颈动脉球囊损伤后血管内皮及AMPK、eNOS表达的影响,探讨其可能的作用机制。方法 32只SD大鼠随机分为假手术组、模型组、颅内消瘀汤低剂量(临床等效量)组和颅内消瘀汤高剂量(临床2倍量)组,除假手术组大鼠仅切开皮肤,其余大鼠均行颈总动脉球囊损伤术以成模。成模第4天灌胃给药,模型组及假手术组给予等体积生理盐水,连续给药14 d后取损伤侧颈总动脉,行HE染色观测血管内皮损伤及内膜增生情况,用免疫组化染色检测eNOS表达水平,并用Western blot检测总AMPK、eNOS蛋白水平。结果颈总动脉球囊损伤模型组大鼠血管内膜增生明显、管壁变厚,颅内消瘀汤低、高剂量组大鼠经治疗后损伤血管增生较模型组轻、管壁较薄;e NOS免疫组化染色:模型组阳性细胞较少,经颅内消瘀汤干预后,低、高剂量2组均有不同程度的表达,与模型组比较差异有统计学意义(P<0.01);Western blot检测结果显示模型组总AMPK、eNOS蛋白表达下降,经颅内消瘀汤干预后,均有不同程度增加,与模型组比较差异有统计学意义(P<0.05或P<0.01)。结论颅内消瘀汤可通过上调AMPK/eNOS信号表达,保护大鼠颈动脉球囊损伤血管内皮的作用。

OBJECTIVE To observe the effect of Lunei Xiaoyu Decoction on vascular endothelial AMPK, eNOS expression in ballon injury model of common carotid artery in rats, and to explore its possible mechanism. METHODS Thirty two SD rats were randomly divided into sham operation group, model group, low dose of Lunei Xiaoyu Decoction（dose in clinical equivalent） group and high dose of Lunei Xiaoyu Decoction（2 times the amount of clinical） group, except the sham operation group rats only skin incision, the other rats underwent carotid artery balloon injury surgery. On the 4th day after modeling, the rats were administrated orally for 14 d, model group and sham operation group were given equal volume of physiological saline. Then all the rats were killed, HE staining observation of vascular endothelial injury and intimal hyperplasia, immunohistochemistry staining was used to detect the expression level of eNOS, and Western-blot was used to detect the expression level of AMPK and eNOS. RESULTS The endometrial hyperplasia and thickness were obvious after carotid artery balloon injury in rats; and after Lunei Xiaoyu Decoction treatment, the rats＇ endometrial hyperplasia and thickness were improved. The expression of AMPKand eNOS was significantly higher in Lunei Xiaoyu Decoction groups compared with the model group（P〈0.05 or P〈0.01）. CONCLUSION Lunei Xiaoyu Decoction can upregulate the expression of AMPK/eNOS signal, in order to protecting vascular endothelial after balloon injured carotid artery in rats.