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目的:研究回心草水提取液及其含药血清对人脐静脉内皮活细胞数、内皮细胞一氧化氮(NO)及一氧化氮合酶(NO合酶)的影响,探讨回心草抗动脉硬化的药理作用机制。方法:采用H2O2建立体外培养的人脐静脉血管内皮细胞(HUVEC)损伤模型。采用MTT法测定回心草提液对H2O2损伤的内皮细胞的吸光度(OD);采用经典Griess Reagent检测各剂量组细胞上清液中NO浓度,采用一氧化氮荧光检测探针DAF-FMDA(3-amino,4-aminomethyl-2′,7′-difluorescein,diacetate)检测细胞内NOS的活性。结果:H2O212.5 mmol.L-1为最合适的细胞刺激浓度;回心草药液与H2O2损伤的血管内皮细胞共同孵育24 h后,2.50 mg.mL-1、3.33 mg.mL-1浓度组与模型组比较有显著差异(P<0.05,P<0.01);回心草水提液作用于内皮细胞,进行NO,NOS含量的测定,其中回心草2.50 mg.mL-1剂量组有显著作用(P<0.05)。结论:12.5 mmol.L-1的H2O2作用24 h,可成功建立内皮细胞氧化应激损伤模型;回心草水提液直接作用于H2O2损伤的内皮细胞,2.50,3.33 mg.mL-1两个浓度组可以减轻内皮细胞的损伤;回心草2.50 mg.mL-1可增加NOS活性,促进NO的分泌。
OBJECTIVE: To investigate the effects of aqueous extracts of Radix edulis and its drug-containing serum on the number of viable cells in human umbilical vein endothelial cells, nitric oxide (NO) and nitric oxide synthase (NO synthase) in endothelial cells, and to investigate the resistance of ricinus. The pharmacological mechanism of atherosclerosis. METHODS: Human umbilical vein endothelial cells (HUVEC) injury model was established with H2O2 in vitro. MTT assay was used to determine the absorbance (OD) of H2O2-damaged endothelial cells. The concentration of NO in cell supernatants was measured by classical Griess Reagent. The nitric oxide fluorescence probe DAF-FMDA was used. -amino, 4-aminomethyl-2’, 7’-difluorescein, diacetate was used to detect the activity of NOS in cells. RESULTS: H2O212.5 mmol.L-1 was the most suitable cell stimulating concentration. The concentration of 2.50 mg.mL-1, 3.33 mg.mL-1 after 24 h incubation of the herbal extracts of H. erythraea with H2O2-injured vascular endothelial cells. There was a significant difference between the model group and the model group (P<0.05, P<0.01). The aqueous extracts of Russulae acted on the endothelial cells, and the contents of NO and NOS were measured. The concentration of the 2.50 mg.mL-1 of R. officinale was significant. Effect (P<0.05). CONCLUSION: The oxidative stress injury model of endothelial cells can be successfully established by the treatment of 12.5 mmol.L-1 H2O2 for 24 h. The water extract of R. officinalis acted directly on H2O2 injured endothelial cells, 2.50, 3.33 mg.mL-1. Concentration group can reduce endothelial cell injury; Concentrate 2.50 mg.mL-1 can increase NOS activity and promote NO secretion.