一株小麦内生链格孢霉代谢产物对兔膝软骨细胞的影响

发布时间：2018-03-25 07:00

  本文选题：大骨节病　切入点：内生真菌　出处：《基因组学与应用生物学》2016年04期

【摘要】：本研究选择分离自青海大骨节病区小麦麦穗中的一株优势内生链格孢霉,探究其代谢产物对兔软骨细胞的影响。通过显微形态学观察和ITS序列分析鉴定菌种;其代谢产物采用75%乙醇提取,然后依次用石油醚、乙酸乙酯、乙醚萃取;methylthiazolyldiphenyl-tetrazoliumbromide(MTT)法检测兔软骨细胞存活率;反转录-聚合酶链式反应(reverse transcription and polymerase chain reaction)检测软骨细胞蛋白聚糖和Ⅱ型胶原蛋白的mRNA表达量;甲苯胺蓝染色法(toluidine blue,TB)和免疫组化染色法(immunological histological chemistry,IHC)分别检测蛋白聚糖和Ⅱ型胶原蛋白的表达。经鉴定实验菌株(编号13-8-A)为互隔链孢霉(Alternaria alternate)。与对照组相比,实验组中的乙醇粗提物能抑制软骨细胞生长,对蛋白聚糖和Ⅱ型胶原蛋白的表达及其mRNA的表达量均有抑制作用;其他萃取分部提取物都能不同程度地损伤软骨细胞,但它们对这两种蛋白的表达及其mRNA表达量的抑制均弱于相同浓度下乙醇粗提物的处理。互隔链孢霉13-8-A代谢产物中应该存在多种可以损伤软骨细胞且可能具有协同作用的物质。
[Abstract]:In this study, a dominant endophytic streptomyces was isolated from the wheat panicle of Kashin-Beck disease area in Qinghai Province, and the effect of its metabolites on rabbit chondrocytes was investigated. The strain was identified by microscopic observation and ITS sequence analysis. The metabolites were extracted by 75% ethanol, followed by petroleum ether, ethyl acetate and methyl thiazolyldiphenyl-tetrazoliumbromide MTT to determine the survival rate of rabbit chondrocytes. Reverse transcription and polymerase chain reactionation was used to detect the mRNA expression of proteoglycan and type 鈪，

本文编号：1662026