龙生蛭胶囊抑制大鼠动脉粥样硬化形成保护血管内皮功能的分子机制研究

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目的研究龙生蛭胶囊对动脉粥样硬化(atherosclerosis,AS)大鼠动脉血管内皮的保护作用,并对其作用机理进行初步探讨。方法正常对照组给予普通饲料,其余大鼠连续给予高脂饲料12周,同时于实验开始时1次性腹腔注射维生素D60万IU·kg~(-1)复制AS模型。于开始造模4周后各组大鼠分别灌胃龙生蛭(LSZ)胶囊低、中、高3个剂量(0.72,1.44,2.88 g·kg~(-1)),辛伐他汀5.5 mg·kg~(-1),正常对照组给予等体积生理盐水。采用生物化学法检测血脂总胆固醇(TC)、甘油三脂(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)水平及一氧化氮(NO)含量;敏感肌张力描计技术检测动脉血管收缩/舒张功能;Western blot检测内皮型一氧化氮合酶(e NOS)及血管细胞黏附分子~(-1)(VCAM~(-1))的蛋白活性及表达;ELASA测定血清白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)的水平。结果与模型组比较,龙生蛭胶囊中、高剂量组均能降低血液中TC、TG、LDL-C并升高HDL-C,降低血清IL-6、TNF-α水平,增加血清NO含量并提高血管内皮e NOS活性(P<0.05,P<0.01);龙生蛭胶囊高剂量组可明显改善高脂饲料诱导的动脉血管环对苯肾上腺素(PE)和5-羟色氨(5-HT)的收缩反应,及对内皮依赖性舒张剂乙酰胆碱(ACh)的舒张反应,并且舒张作用可被N-硝基-L-精氨酸甲酯(L-NAME)阻断(P<0.05,P<0.01)。实验结果还发现龙生蛭胶囊中、高剂量组均能降低由高脂诱导的VCAM~(-1)表达升高(P<0.05,P<0.01)。结论龙生蛭胶囊可显著性降低AS大鼠血脂水平;改善大鼠血管内皮依赖性血管舒张功能;抑制系统炎性反应,其机制可能与上调e NOS活性、抑制VCAM~(-1)表达相关。表明龙生蛭胶囊不仅可用于防治脑梗的发生,还对AS早期病变具有良好的治疗作用。 Objective To study the protective effect of “Longshengzhudi Capsule” on the vascular endothelium of atherosclerosis (AS) rats and its mechanism of action. Methods The normal control group was given normal diet, and the other rats were given continuous high fat diet for 12 weeks. At the same time, the rats were injected intraperitoneally with 6 million IU · kg -1 of vitamin D for AS model at the beginning of the experiment. Four weeks after the beginning of modeling, the rats in each group were given low, medium and high doses of 0.32, 1.44 and 2.88 g · kg -1 of LSZ capsule respectively, simvastatin 5.5 mg · Kg ~ (-1), the normal control group was given equal volume of normal saline. The levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) and nitric oxide (NO) The contractile / diastolic function of arterial vessels was detected by sensitive muscle tension meter. The protein activities of endothelial nitric oxide synthase (eNOS) and vascular cell adhesion molecule (-1) (VCAM -1) were detected by Western blot and The levels of serum IL-6 and TNF-α were measured by ELASA. Results Compared with the model group, Longshengzhidong Capsule could lower TC, TG and LDL-C in blood and increase HDL-C, lower the level of serum IL-6 and TNF-α and increase the content of serum NO (P <0.05, P <0.01). The high-dose Longshenzhili Capsule group could significantly improve the effect of high-fat diet on arterial blood vessel rings of phenylephrine (PE) and 5- HT) and the relaxation response to acetylcholine (ACh), an endothelium-dependent relaxation agent, and the diastolic action was blocked by N-nitro-L-arginine methyl ester (L-NAME) P <0.01). The experimental results also found that both Longshengzhidi capsule and high-dose group can reduce the high-fat-induced VCAM -1 expression (P <0.05, P <0.01). Conclusion Longshengzhirong Capsule can significantly reduce the blood lipid level of AS rats, improve vascular endothelium-dependent vasodilatation and inhibit systemic inflammatory response in rats, which may be related to up-regulation of eNOS activity and inhibition of VCAM-1 expression . That Longsheng Leech Capsule can be used not only to prevent the occurrence of cerebral infarction, AS also has a good early treatment of disease.
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