不同硒浓度培养的一株链格孢霉产物对兔软骨细胞的影响  

Effect of Products Extracted from an Alternaria alternate Strain Incubated with Different Selenium Concentrations on Rabbit Chondrocytes

作　　者：张枫[1,2] 张燕 赵玉濛[1] 胡锦涛 殷红 Zhang Feng;Zhang Yan;Zhao Yumeng;Hu Jintao;Yin Hong(The College of Life Science,Northwest University,Xi'an,710069;Hanzhong Center for Disease Control and Prevention,Hanzhong,723000;Baoji Center for Disease Control and Prevention,Baoji,721006;Key Laboratory of Resource Biology and Biotechnology in Western China(Northwest University),Ministry of Education,Xi'an,710069)

机构地区：[1]西北大学生命科学学院,西安710069 [2]汉中市疾病预防控制中心,汉中723000 [3]宝鸡市疾病预防控制中心,宝鸡721006 [4]西部资源生物与现代生物技术教育部重点实验室,西安710069

出　　处：《基因组学与应用生物学》2019年第8期3566-3571,共6页Genomics and Applied Biology

基　　金：陕西省卫生厅疾控项目(2010/2012);陕西省自然科学基础研究计划项目(2014JM4170)共同资助

摘　　要：本实验研究了加硒培养的一株链格孢霉(Alternaria alternate)菌株的产物对兔软骨细胞毒损作用的影响。采用合成培养液,添加不同浓度硒元素后进行摇瓶培养链格孢霉菌株;发酵产物用75%乙醇提取后减压浓缩;将不同硒处理的发酵产物提取物加入兔软骨细胞培养体系,采用四氮唑蓝(MTT)法测定软骨细胞的生长速率,甲苯胺蓝染色法检测其蛋白聚糖的表达,免疫组化法检测Ⅱ型胶原表达量,半定量RT-PCR法检测细胞蛋白聚糖和Ⅱ型胶原mRNA的表达。结果显示,在实验所设浓度范围内,加硒对该链格孢霉菌株的生长有一定影响,但不显著。加不同浓度硒培养所得发酵产物提取物对兔软骨细胞生长速率的影响各异,多表现为抑制作用。菌株发酵产物的提取物对Ⅱ型胶原和蛋白聚糖mRNA的表达量有些微促进作用,尤其是加一定浓度硒的处理;但对这两种蛋白的表达都有抑制作用,不过和未加硒的处理相比,加一定浓度硒培养所得提取物的抑制效果有所减轻。本研究结果表明,硒对该链格孢霉菌株的生长有一定影响;加不同浓度硒培养的菌株提取物对兔软骨细胞损伤有关的指标影响不同;说明环境硒水平-真菌产物毒性-软骨细胞损伤之间可能存在一定关系,值得进一步研究。We studied the effects of the product of an Alternaria alternate strain cultured with selenium on rabbit knee cartilage cells. The A. alternate strain was cultured with shaking in flasks containing chemically defined medium and various concentrations of selenium. Fermentation products were extracted with 75% ethanol and concentrated by decompression. Extracted fermentation products were then incubated with rabbit cartilage cell cultures. Growth rate of cartilage cells was determined by tetrazolium blue(MTT) assay, the expression of proteoglycan was detected by toluidine blue staining, the expression of type Ⅱ collagen by immunohistochemically staining, the proteoglycan and type Ⅱ collagen m RNA expression levels were detected by semi-quantitative RT-PCR. Results showed that the addition of selenium resulted in minor changes in mycelia growth of the A. alternate strain. Effects of the extracted fermentation products cultured with different concentrations of selenium on the growth rate of cartilage cells mainly inhibitory. Although both the proteoglycan and type Ⅱ collagen mRNA expression levels were elevated to some extent, their protein levels were inhibited to varying degrees compared with untreated control cells. However, the inhibitory effect was relatively low when the fungal culture medium was supplemented with certain selenium concentrations. The results indicated that selenium had a certain effect on the mycelial growth of the A. alternate strain. Indices of cartilage cell injury were altered by the fermentation products of A. alternate treated with different selenium concentrations. Further study of the relationship between selenium, toxic metabolites produced by fungi, and cartilage cell damage is warranted.

关 键 词：硒 链格孢霉 发酵产物 软骨细胞 

分 类 号：R28[医药卫生—中药学]