青海大骨节病区小麦内生镰刀菌发酵产物和硒对兔膝软骨细胞的影响

发布时间：2018-05-07 19:42

本文选题：大骨节病 + 硒元素　；参考：《西北大学》2015年硕士论文

【摘要】：大骨节病是一种地方性、变形性骨关节病,对儿童的生长和发育有严重的影响,具有相当高的患病率和致残率,目前其病因假说主要集中于环境因素探讨,包括真菌毒素中毒说、低硒学说等,具体致病机制目前还不清楚。本实验采用从青海大骨节病区穗期小麦中分离出的优势内生镰刀菌,联合硒对镰刀菌的影响,将不同硒水平下培养镰刀菌得到的发酵产物作用于体外培养的软骨细胞,通过MTT法、甲苯胺蓝和免疫组化染色法、RT-PCR法检测软骨细胞病变的相关生理指标。结果显示,三株镰刀菌发酵产物均能明显降低细胞存活率；同时也显著减少Ⅱ型胶原和蛋白聚糖的合成及抑制其mRNA的转录水平；适宜浓度硒条件下培养菌株所获发酵产物对蛋白聚糖和Ⅱ型胶原蛋白合成的抑制作用比无硒的小。对于5-5-1菌株和6-5-1菌株,当硒浓度高于0.5mg/L时抑制反而增大；对于1-8-B菌株,硒浓度高于0.5mg/L时,相比较低硒浓度时,发酵产物对这两种蛋白合成的抑制作用大,但相比无硒的抑制作用略小。取上述三株菌中产T-2毒素的5-5-1菌株,将不同浓度的T-2毒素和5-5-1菌株发酵产物作用于体外培养的兔软骨细胞。实验结果显示,二者均能降低软骨细胞存活率,明显抑制蛋白聚糖和Ⅱ型胶原的合成及其mRNA的转录水平。相比纯T-2毒素,含有类似T-2毒素含量的5-5-1菌株发酵产物对细胞存活率、抑制蛋白聚糖和Ⅱ型胶原的合成及mRNA的转录水平的抑制作用更大。结果显示三株实验菌的发酵产物都能够使软骨细胞发生类似大骨节病的病变特征,相比无硒条件培养的镰刀菌,添加适宜浓度硒培养菌株得到的发酵产物对软骨细胞相关指标的抑制作用相对较小,且含有同样T-2毒素含量的5-5-1菌株发酵产物比纯T-2毒素对软骨细胞的抑制作用更大,结果提示,适宜浓度硒可能抑制了菌株的产毒能力,从而减轻其发酵产物对细胞的毒性作用；镰刀菌毒素对软骨细胞的毒害可能是多种真菌毒素综合作用的结果。
[Abstract]:Kashin Beck disease is a local, deformable osteoarthropathy, which has a serious influence on the growth and development of children. It has a high morbidity and disability rate. At present, the hypothesis of etiology is mainly focused on the environmental factors, including the theory of mycotoxin poisoning, the theory of low selenium and so on. The pathogenesis of the disease is not yet clear. This experiment was used from Qinghai. The predominant endophytic Fusarium in the ear stage of the Kaschin Beck disease area, combined with the effect of selenium on the Fusarium, acted on the cultured chondrocytes cultured in vitro by the fermentation products obtained from different selenium levels, by MTT method, toluidine blue and immunohistochemical staining, and RT-PCR method to detect the related physiological indexes of chondrocyte disease. The results showed that the fermentation products of three Fusarium strains could obviously reduce the cell survival rate, and also significantly reduce the synthesis of type II collagen and proteoglycan and inhibit the transcriptional level of mRNA. The inhibitory effect of fermentation products on the synthesis of proteoglycan and type II collagen egg white under the suitable concentration of selenium condition is smaller than that of the selenium free. For 5- 5-1 strains and 6-5-1 strains, when the selenium concentration is higher than 0.5mg/L, the inhibition increases; for the 1-8-B strain, the selenium concentration is higher than the 0.5mg/L, when the selenium concentration is higher than the selenium concentration, the inhibition effect of the fermentation product on the two proteins is large, but the inhibition effect is slightly smaller than that of the selenium free. The 5-5-1 strain of the T-2 toxin of the three strains of the above three strains will have different concentrations. The fermentation products of T-2 toxin and 5-5-1 strain acted on the rabbit chondrocytes cultured in vitro. The results showed that the two groups could reduce the survival rate of cartilage cells, inhibit the synthesis of proteoglycan and type II collagen and the transcriptional level of mRNA. Compared with pure T-2 toxin, the fermentation products containing 5-5-1 strain, similar to T-2 toxin, survived the cell survival. Rate, inhibition of the synthesis of proteoglycan and type II collagen and the inhibition of the transcriptional level of mRNA, the results showed that the fermentation products of three strains of experimental bacteria could make chondrocytes similar to the disease characteristics of Kaschin Beck disease. Compared with the selenium free Fusarium, the fermentation products with suitable concentration selenium culture strain were fine to the cartilage. The inhibitory effects of cell related indexes were relatively small, and the fermentation products containing the same T-2 toxin content were more inhibited than the pure T-2 toxin on chondrocytes. The results suggest that the suitable concentration of selenium may inhibit the toxicity of the strain, thus reducing the toxicity of the fermentation products to the cells, and the Fusarium toxin is fine to the cartilage. Cytotoxicity may be the result of the combined action of various mycotoxins.

【学位授予单位】：西北大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R684.1

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