靈芝多糖拮抗前列腺素e2對小鼠脾細(xì)胞ifn-γ和tnf-α-全文預(yù)覽
【正文】 (20 μmol/L)。靈芝多糖拮抗PGE2對小鼠混合淋巴細(xì)胞培養(yǎng)反應(yīng)中脾細(xì)胞IFNγ mRNA表達(dá)的抑制作用 5: PGE2 (40 μmol/L)。1: Control。 PGE2 inhibits murine splenocyte IFNγ mRNA expression in mixed lymphocyte culture reaction (MLR)圖1 PGE2對小鼠混合淋巴細(xì)胞培養(yǎng)反應(yīng)中脾細(xì)胞IFNγ mRNA表達(dá)的抑制作用 不同濃度PGE2連續(xù)作用4 h后，MLR中小鼠脾細(xì)胞IFNγ mRNA表達(dá)受到不同程度的抑制，隨著PGE2濃度的增加，抑制作用不斷增強(qiáng)。PGE2對混合淋巴細(xì)胞培養(yǎng)反應(yīng)(MLR)中IFNγ mRNA表達(dá)的抑制作用2相應(yīng)βactin擴(kuò)增產(chǎn)物熒光強(qiáng)度值)100%。循環(huán)完畢，72 ℃延伸5 min。ggg gtg ttg aag gtc tca aa339。PCR擴(kuò)增產(chǎn)物為422 bp。小鼠TNFα上游引物：539。ttt gag gtc aac aac cca ca339。PCR擴(kuò)增吸取上述混合液20 μl加入AccuPower RT PreMix管中，振蕩混勻，快速離心數(shù)秒，置PCR循環(huán)儀中反應(yīng)。然后固定PGE2濃度為20 μmol/L，在其中再加入不同濃度的靈芝多糖(100、200 mg/L)，觀察靈芝多糖拮抗PGE2對IFNγ或TNFα mRNA表達(dá)的抑制作用，共設(shè)5組，即：空白對照組(加等體積培養(yǎng)液)，靈芝多糖(200 mg/L)促進(jìn)表達(dá)組，PGE2(20 μmol/L)抑制表達(dá)組，PGE2(20 μmol/L)+靈芝多糖(100 mg/L)組，PGE2(20 μmol)+靈芝多糖(200 mg/L)組。實驗方案儀器靈芝多糖參照文獻(xiàn)[6]方法提取，為多糖組分，呈淺棕色，易溶于水，相對分子質(zhì)量7000～9000。DEPC、PGE2均為Sigma產(chǎn)品。動物1 材料和方法其中，多糖是靈芝中主要的生物活性成分之一。 mRNA收稿日期：20051107基金項目：廣東省自然科學(xué)基金(04020361)Sponsored by Natural Science Foundation of Guangdong Province (04020361)作者簡介：張文章編號：16734254(2006)06078004Ganoderma polysaccharides antagonize prostaglandin E2induced suppression of murine splenocyte IFNγ and TNFα mRNA expressionZHANG Qun；LEI Linsheng；ZHU Zhengguang；YU Chuanlin；WU ShuguangInstitute of Pharmaceutical Sciences, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, ChinaAbstract: Objective To determine if Ganoderma polysaccharides can antagonize prostaglandin E2 (PGE2)induced suppression of murine splenocyte interferon γ (IFNγ) and tumor necrosis factor α (TNFα) mRNA expression. Methods Mixed lymphocyte culture reaction was used as the experimental model. The expressions levels of IFNγ and TNFα mRNA were measured by semiquantitative reverse transcriptasepolymerase chain reaction (RTPCR). Results After the cultures were treated with PGE2 for 4 h, IFNγ mRNA expression was reduced as pared with the control, which was especially obvious when PGE2 concentrations exceeded 10 μmol/L (P). Ganoderma polysaccharides above
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