摘要
目的探讨香菇多糖(Lentinan)对脓毒症引起的小鼠急性肺损伤的保护作用。方法采用噻唑蓝(MTT)法检测香菇多糖对RAW264.7细胞生长的影响。通过脂多糖LPS(Lipopolysaccharide)诱导RAW264.7细胞产生炎症反应,并检测培养基中白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)和一氧化氮(NO)的浓度,Western blot法检测细胞中环氧化酶(COX-2)、一氧化氮合酶(iNOS)、TLR4及NF-κB p65蛋白的表达水平。体内实验苏木精-伊红染色法(HE)观察小鼠肺组织的病理改变,免疫组化检测p65蛋白表达水平。结果 50μM香菇多糖可明显抑制炎症因子IL-6、IL-1β、NO的释放,且随香菇多糖浓度升高,炎症因子的释放量减少更显著(均P<0.05);100μM香菇多糖能下调COX-2、iNOS、TLR4及NF-κB p65蛋白表达水平。体内实验HE染色结果表明:对照组细胞完整,细胞核完整,细胞界限清晰,炎性细胞很少或不可见;而模型组中细胞核染不完整,细胞界限不清,有大量炎性细胞浸润;100μM香菇多糖组明显改善了模型组上述状况,表现出对小鼠肺部损伤的保护作用;免疫组化结果显示模型组p65蛋白表达较对照组升高,而100μM香菇多糖可明显下调p65蛋白表达水平。结论香菇多糖通过抑制IL-6、IL-1β和NO信号分子,下调iNOS、COX-2、TLR4、NF-κB p65蛋白的表达而发挥抗炎作用,对脓毒症诱导急性肺损伤有保护作用。
OBJECTIVE To investigate the protective effect of lentinan on the acute lung injury induced by sepsis in a mice model.METHODS The influence of lentinan on growth of RAW264.7 cell was detected by using methyl thiazolyl tetrazolium（MTT）.The RAW264.7 cell was induced for inflammatory reaction by lipopolysaccharide（LPS）,the concentrations of interleukin-6（IL-6）,interleukin-1β（IL-1β）and nitric oxide（NO）in medium were detected,the levels of expression of cyclooxygenase（COX-2）,nitric oxide synthase（iNOS）,TLR4 and NF-κB p65 in the cell were detected with the use of Western blot.The pathologic change of lung tissues of the mice was observed by means of in vivo experiment hematoxylin-eosin staining（HE）,and the level of expression of p65 protein was detected by using immunohistochemistry assay.RESULTS 50μM lentinan could obviously inhibit the release of inflammatory factors IL-6,IL-1βand NO,and released amount of the inflammatory factors was reduced more significantly with the increase of concentration of lentinan（all P〈0.05）;100μM lentinan could down-regulate the levels of expression of COX-2,iNOS,TLR4 and NF-κB p65.The result of the in vivo experiment HE staining indicated that the cells and cell nucleus were complete in the control group,with cellular boundaries clear,and there were few inflammatory cells or they could not be seen;the cell nucleus were incomplete in the model group,with the cellular boundaries unclear,and there were large number of inflammatory cells of infiltration.100μM lentinan significantly improved the above condition in the model group,showing the protective effect on the lung injury of the mice.The result of the immunohistochemistry assay showed that the level of expression of p65 protein was higher in the mode group than in the control group,however,100μM lentinan could remarkably down-regulate the level of expression of p65 protein.CONCLUSION Lentinan plays the anti-inflammatory effect by inhibiting the IL-6,IL-1βand NO signal molecule and down-reg
作者
章静
杨成伟
王云
汤文建
ZHANG Jing;YANG Cheng-wei;WANG Yun;TANG Wen-jian(Anhui Prevention and Treatment Center for Occupational Disease, Anhui No.2 Province People＇s Hospital, Hefei, Anhui 230022, Chin)
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2018年第10期1463-1468,共6页
Chinese Journal of Nosocomiology
基金
安徽省教育厅科研基金资助项目(12925KJ2017b03)
安徽省"十三五"医疗卫生重点培育实验室基金资助项目(皖卫科教2017-30)
