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目的:探讨参莲(SL)提取物在炎症调节中发挥的治疗性作用及其机制。方法:分别以小鼠巨噬细胞Raw264.7和小鼠腹腔巨噬细胞为实验对象,利用脂多糖(LPS)诱导炎症模型,SL提取物低、中、高剂量(5,10,20 mg·L~(-1))药物处理24 h,另设空白组,使用酶联免疫吸附测定(ELISA)法及实时荧光定量PCR(Real-time PCR)检测小鼠Raw264.7及腹腔巨噬细胞肿瘤坏死因子-α(TNF-α),白细胞介素-1β(IL-1β)含量及mRNA表达;使用蛋白质免疫印迹法(Western blot)对小鼠Raw264.7核转录因子-κB(NF-κB)信号通路中的p65及磷酸化p65(p-p65)蛋白表达情况进行检测,采用免疫荧光法对小鼠Raw264.7及腹腔巨噬细胞NF-κB p65分子进行亚细胞定位观察SL提取物是否能够影响其入核。结果:与空白组比较,模型组小鼠Raw264.7及腹腔巨噬细胞TNF-α,IL-1β含量及mRNA表达均显著升高,小鼠Raw264.7 p-p65/p65蛋白表达显著升高(P<0.01),免疫荧光观察显示模型组Raw264.7及腹腔巨噬细胞均可见明显入核行为;与模型组比较,SL提取物低、中、高剂量组明显降低小鼠Raw264.7及腹腔巨噬细胞TNF-α,IL-1β含量及mRNA表达(P<0.05,P<0.01),显著降低小鼠Raw264.7及p-p65蛋白(P<0.01),SL提取物中剂量组可明显减弱Raw264.7及腹腔巨噬细胞入核行为。结论:SL提取物能够抑制巨噬细胞的炎症反应,其机制可能与影响巨噬细胞炎性因子的分泌及NF-κB信号通路有关。
Objective: To investigate the therapeutic effect and mechanism of the extract of Senate lotus (SL) on inflammation. Methods: The mouse macrophages Raw264.7 and mouse peritoneal macrophages were used as experimental subjects. Lipopolysaccharide (LPS) -induced inflammation model was used. SL extract was administered at low, medium and high doses (5, 10, 20 mg · L ~ (-1)) for 24 h, and another blank group. The expression of Raw264.7 and peritoneal macrophage tumor were detected by ELISA and Real-time PCR (TNF-α), interleukin-1β (IL-1β) and mRNA expression were detected by Western blotting.Western blotting was used to detect the expression of nuclear factor-kappa B (NF- (P-p65) in the signal pathways of mice were detected by immunofluorescence method for the mouse Raw264.7 and peritoneal macrophages NF-κB p65 molecules subcellular localization SL extract whether Affect its entry into the nuclear. Results: Compared with the blank group, the content of Raw264.7 and the expression of TNF-α and IL-1β in peritoneal macrophages in model group were significantly increased, and the protein expression of Raw264.7 p-p65 / p65 in mouse was significantly increased (P <0.01). The results of immunofluorescence showed that the expression of NF-κB in Raw264.7 and peritoneal macrophages was significantly increased in the model group. Compared with the model group, the low, medium and high dose SL extracts significantly reduced the expression of Raw264.7 and The levels of TNF-α, IL-1β and mRNA in peritoneal macrophages were significantly decreased (P <0.05, P <0.01), and the protein levels of Raw264.7 and p-p65 in mice were significantly decreased Significantly reduced Raw264.7 and peritoneal macrophages into the nucleus. CONCLUSION: SL extract can inhibit the inflammatory response of macrophages, and its mechanism may be related to the secretion of inflammatory cytokines and the NF-κB signaling pathway.