红鱼眼不同提取物含药血清对脂多糖诱导RAW264.7细胞炎症的影响

Drug Serum From Different Extracts of Phyllanthus Reticulatus Poir.on Inflammatory Factors in LPS-induced Macrophage RAW264.7

[目的]研究红鱼眼不同极性部位提取物含药血清对脂多糖(LPS)诱导的RAW264.7细胞炎症的影响。[方法]采用噻唑蓝(MTT)法检测红鱼眼不同极性部位提取物含药血清对RAW264.7细胞活力的影响;采用脂多糖(LPS)诱导RAW264.7细胞建立炎症模型,采用MTT法检测红鱼眼不同极性部位提取物含药血清对炎症细胞增殖的影响,并用酶联免疫吸附测定(ELISA)法检测细胞上清液中一氧化氮(NO)、白介素1β(1L-1β)、白介素6(1L-6)及肿瘤坏死因子-α(TNF-α)的含量。[结果]在6~12 h培养时间内,除红鱼眼水提取物含药血清外,其余不同极性部位提取物含药血清对RAW264.7细胞活力均无明显影响;与LPS模型组比较,红鱼眼不同极性部位提取物组各培养时间段吸光度(OD)值和细胞存活率明显提高(P<0.01);LPS刺激细胞后可引起NO、IL-1β、IL-6、TNF-α的高表达(P<0.05或P<0.01),经红鱼眼不同极性部位提取物含药血清干预后,乙酸乙酯提取物组、石油醚提取物组、水提取物组的NO、IL-1β、IL-6、TNF-α含量显著下降(P<0.01);80%乙醇提取物组的IL-6、IL-1β含量显著下降(P<0.01);水饱和正丁醇提取物组的IL-1β、NO、TNF-α含量显著下降(P<0.01)。[结论]红鱼眼不同极性部位(水部位除外)提取物含药血清在12 h内对RAW264.7细胞活力无影响,但对LPS诱导的RAW264.7炎症细胞具有一定的保护作用,其机制可能是通过抑制LPS诱导巨噬细胞炎症反应而发挥抗炎作用。

[Objective]To study the effect of drug serum form different extracts of Phyllanthus reticulatus Poir.on inflammatory factors on lipopolysaccharide(LPS)-induced RAW264.7 inflammation.[Methods]To detect the effect of different extracts of Phyllanthus reticulatus Poir.drug serum on the proliferation activity of RAW264.7 cells by MTT method.LPS was applied to induce RAW264.7 cells to establish an inflammatory model.MTT assay was used to detect the effect of serum containing different extracts of redfish eyes on the activity of inflammatory cells.The contents of nitric oxide(NO),interleukin 1β(1L-1β),interleukin 6(1L-6)and tumor necrosis factor-α(TNF-α)in cell supernatant were detected by enzyme-linked immunosorbent assay(ELISA).[Results]Within the incubation time range of 6~12 hours,the drug serum form different extracts of Phyllanthus reticulatus Poir.did not show significant cell death to RAW264.7 cells.Compared with the LPS model group,the OD value and cell survival rate of the extracts of different polar parts of Phyllanthus reticulatus Poir.were significantly increased(P<0.01),and the expression of NO,IL-1β,IL-6 and TNF-αwas increased after LPS stimulation(P<0.05 or P<0.01).After the intervention of medicated serum containing extracts from different polar parts of Phyllanthus reticulatus Poir.the contents of NO,IL-1β,IL-6 and TNF-αin ethyl acetate extract group,petroleum ether extract group and water extract group were significantly decreased(P<0.01).The content of IL-6 and IL-1βin 80%ethanol extract group decreased significantly(P<0.01).The contents of IL-1β,NO and TNF-αin the water-saturated n-butanol extract group were significantly decreased(P<0.01)[.Conclusion]The drug serum from different extracts of Phyllanthus reticulatus Poir.has no effect on the viability of RAW264.7 cells within 12 hours,and it has a certain protective effect on LPS-induced RAW264.7 inflammatory cells,and its mechanism may play an anti-inflammatory role by inhibiting LPS-induced macrophage inflammatory response.