阿里红多糖对Aβ_(25-35)诱导的小胶质细胞炎症反应的影响

Effects of Fomes officinais Ames polysaccharides on Aβ_(25-35) induced inflammatory response of microglia

目的探讨阿里红多糖(Fomes Officinals polysaccharide,FOPS)及多糖组分(FOP-a)对Aβ_(25-35)诱导的BV-2小胶质细胞炎症反应的抑制作用。方法采用Aβ_(25-35)诱导BV-2小胶质细胞活化建立阿尔茨海默病(alzheimerdisease,AD)炎症细胞模型,将BV-2细胞分为空白组、炎症模型组(加40μg/mLAβ_(25-35))、不同浓度的FOPS及FOP-a干预组(6.25、12.5、25μg/mL)。在倒置显微镜下观察细胞形态,酶联免疫法(enzyme linked immunosorbentassay,ELISA)检测细胞上清中IL-6及单核细胞趋化因子(MCP-1)分泌量;Westernblotting法测定NF-κB及IκBα蛋白表达量。结果 Aβ_(25-35)诱导后小胶质细胞出现阿米巴样状态,通过不同剂量FOPS及FOP-a干预,能明显改善Aβ_(25-35)对BV-2细胞造成的损伤,与模型组比较, FOPS及FOP-a可提高细胞存活率。ELISA实验结果显示与空白组比较,模型组IL-6及MCP-1的分泌量增加,与模型组比较,FOPS及FOP-a均能减少IL-6及MCP-1的分泌量,差异有统计学意义(P<0.05)。Western blotting结果显示,与空白组比较,模型组细胞上调NF-κB-p65蛋白表达、减少IκBα的蛋白表达,与模型组比较FOPS及FOP-a均能减少NF-κB-p65和上调IκBα的蛋白表达。结论阿里红粗多糖及多糖组分均能减轻Aβ_(25-35)诱导的BV-2细胞的炎症作用,其中FOPS浓度为12.5μg/mL、FOP-a浓度为25μg/mL时效果最佳。

Objective To investigate the inhibitory effects of Fomes officinais Ames polysaccharides（FOPS） and polysaccharides components （FOP-a） on the inflammatory response in BV-2 microglial cells induced by Aβ25-35. Methods Alzheimer＇s disease （AD） inflammatory cell model was established by induction of BV-2 microglia activation by Aβ25-35.BV-2 cells were divided into blank group, inflammatory model group （40 μg/mL Aβ25-35）, different concentrations of FOPS and FOP-a intervention group （6.25, 12.5, 25 μg/mL）. The morphology of the cells was observed under an inverted microscope, the secretion of IL-6 and monocyte chemotactic factor （MCP-1） in the supernatant of the cells was detected by enzyme linked immunosorbent assay （ELISA）, and NF-κB and IκBα protein expression were detected by Western blotting. Results After Aβ25-35 induction, microglia showed amoebic-like state. Different doses of FOPS and FOP-a could significantly improve the damage of BV-2 cells induced by Aβ25-35. Compared with the model group, FOPS and FOP-a could increase cell survival rate. The results of ELISA showed that the secretion of IL-6 and MCP-1 increased in the model group compared with the blank group; compared with the model group, both FOPS and FOP-a could reduce the secretion of IL-6 and MCP-1, and the difference was statistically significant （P〈0.05）.Western blotting results showed that compared with the blank group, the model group could up-regulate the expression of NF-κB-p65 protein and decrease the protein expression of IκBα. Both FOPS and FOP-a could reduce NF-κB-p65 and up-regulate IκBα protein expression compared with the model group. Conclusion Polysaccharides components FOPS and FOP-a from Fomes officinais Ames can alleviate the inflammatory effects of Aβ25-35-induced BV-2 cells, and the best effect is when the FOPS concentration is 12.5 μg/mL and the FOP-a concentration is 25 μg/mL.