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目的观察粉防己碱(tetrandrine,Tet)对脂多糖(LPS)诱导RAW264.7细胞炎症模型促炎细胞因子和抗炎细胞因子的影响。方法 LPS(1μg/mL)刺激生长良好的RAW264.7细胞,建立细胞炎症模型。四甲基偶氮唑盐(MTT)法检测不同浓度Tet对RAW264.7细胞增殖的影响,激光共聚焦显微镜观察Tet对细胞核因子-κB(NF-κB)核转运的作用,酶联免疫吸附试验(ELISA)检测细胞上清中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)的变化。结果当Tet浓度<1μmol/L时,单独或与1μg/mL LPS共培养,均对RAW264.7细胞生长无影响;而当Tet浓度>10μmol/L时,单独或与LPS共培养均表现出明显的细胞生长抑制效应;Tet可使IL-6、TNF-α的释放受到抑制,相反可促进IL-10的表达。激光共聚焦显微镜观察Tet大剂量组细胞核红染的阳性细胞显著减少,以阴性细胞(红染p65亚基主要集中在胞浆部位,使胞浆染色较深,胞核染色较浅,整个细胞成空泡状)为主。结论 Tet可以抑制LPS所致的RAW264.7细胞炎症反应,其抗炎作用与抑制NF-κB活化,进一步减少炎性细胞因子IL-6、TNF-α的产生,并促进抗炎细胞因子IL-10的表达等有关。
Objective To investigate the effect of tetrandrine (Tet) on pro-inflammatory cytokines and anti-inflammatory cytokines induced by lipopolysaccharide (LPS) in RAW264.7 cells. Methods LPS (1μg / mL) stimulated the growth of well RAW264.7 cells and established a model of cellular inflammation. MTT assay was used to detect the effect of different concentrations of Tet on the proliferation of RAW264.7 cells. The effect of Tet on NF-κB nuclear translocation was observed by laser scanning confocal microscopy. (IL-6), tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in supernatants were detected by ELISA. Results When Tet concentration was less than 1μmol / L, either alone or in combination with 1μg / mL LPS had no effect on the growth of RAW264.7 cells. When the concentration of Tet> 10μmol / L, either alone or co-cultured with LPS Tet can inhibit the release of IL-6 and TNF-α, and on the contrary, promote the expression of IL-10. Laser confocal microscopy showed that there was a significant decrease in the number of positive cells in the Tet nucleus in the Tet high-dose group. Negative cells (the red staining of the p65 subunit mainly concentrated in the cytoplasm, deep staining of the cytoplasm, Bubble-shaped) based. CONCLUSION: Tet can inhibit the inflammatory response induced by LPS in RAW264.7 cells, and its anti-inflammatory effect and NF-κB activation further reduce the production of inflammatory cytokines IL-6 and TNF-α and promote the anti-inflammatory cytokines IL- 10 expression and so on.