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目的研究四妙勇安汤含药血清对脂多糖(LPS)诱导的小鼠腹腔巨噬细胞释放NO、IL-1β表达的影响,初步探讨其抗动脉粥样硬化(AS)的作用机制。方法制备含药血清选用清洁级雄性SD大鼠50只,随机分为大、中、小剂量给药组,另设一个正常对照组、一个模型组,各10只。制备腹腔巨噬细胞选用清洁级KM小鼠30只。采用大、中、小剂量四妙勇安汤水煎剂灌胃干预SD大鼠,制备含药血清;经MTT法观察含药血清对细胞增殖的影响后,选择各剂量浓度为20%的含药血清干预经LPS活化的巨噬细胞,采用Griess法检测NO释放量;ELISA法检测细胞上清液中IL-1β含量。结果四妙勇安汤含药血清在5%~20%浓度范围内,与相同浓度正常对照组血清OD值比较,差异无统计学意义(P>0.05)。LPS刺激细胞后,模型组NO分泌量大于正常对照组,差异有统计学意义(P<0.05)。四妙勇安汤含药血清各剂量组NO分泌量小于模型组,差异有统计学意义(P<0.05)。用LPS刺激巨噬细胞后,模型组IL-1β分泌量大于正常对照组,差异有统计学意义(P<0.05)。四妙勇安汤含药血清各剂量组作用于细胞后,IL-1β的分泌量小于模型组,差异有统计学意义(P<0.05)。结论四妙勇安汤含药血清能够有效抑制LPS诱导的巨噬细胞炎性反应,这可能是该方防治AS及免疫炎性疾病的机制之一。
Objective To study the effect of Sijiaoyong’an Decoction-containing serum on lipopolysaccharide (LPS) -induced NO and IL-1β expression in mouse peritoneal macrophages and to explore its anti-atherosclerosis (AS) mechanism. Methods Fifty clean male Sprague-Dawley rats were randomly divided into large, medium and small dose groups, and another one normal control group and one model group, 10 rats each. Preparation of peritoneal macrophages use clean KM mice 30. Small, medium and small doses of four Miaoyongan decoction intragastric administration intervention SD rats prepared serum containing; MTT assay of drug-containing serum on cell proliferation, the choice of the dose of 20% of the drug Serum intervention of macrophages activated by LPS, NO release detected by Griess method; ELISA assay of IL-1β content in the cell supernatant. Results There was no significant difference in the serum OD value between the Simiao Yongan Decoction serum and the normal control group at the same concentration of 5% ~ 20% (P> 0.05). After stimulated by LPS, NO secretion in the model group was greater than that in the normal control group, with statistical significance (P <0.05). The quantity of NO secreted by each dose group of Si Miaoyong An Decoction serum was lower than that of model group, the difference was statistically significant (P <0.05). After stimulation of macrophages with LPS, the secretion of IL-1β in the model group was greater than that of the normal control group, the difference was statistically significant (P <0.05). Compared with the model group, the secretion of IL-1β in each dose group of SiMaoYongAnTang containing medicine serum had a significant difference (P <0.05). Conclusion SiMaoYongAnTang serum can effectively inhibit LPS-induced macrophage inflammatory reaction, which may be one of the mechanisms of prevention and treatment of AS and immune inflammatory diseases.