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小麦内生镰刀菌5-5-1菌株是从青海大骨节病区穗期小麦籽粒中分离的一株优势内生镰刀菌,本试验通过采用干重法测定菌丝体生长量;MTT(methylthiazolyldiphenyl-tetrazoliumbromide)法检测软骨细胞存活率;甲苯胺蓝染色法和免疫组化染色法分别检测兔膝软骨细胞内蛋白聚糖(aggrecan)和Ⅱ型胶原蛋白(collagenⅡ)的表达;逆转录-聚合酶链式反应(RT-PCR)法检测软骨细胞内蛋白聚糖和Ⅱ型胶原蛋白m RNA的表达水平,探讨了硒对5-5-1菌株生长的影响及不同硒水平下培养该菌株获得的发酵产物对兔膝软骨细胞有关指标的影响。结果表明,硒浓度在0.05~0.5 mg/L的范围内,对5-5-1菌株生长有促进作用,且呈浓度效应;低浓度硒条件下培养的该菌株发酵产物对软骨细胞蛋白聚糖和Ⅱ型胶原蛋白的合成及其m RNA表达的抑制作用相对较小。由此推断适宜浓度硒具有促进镰刀菌生长的作用,但同时能够减轻其发酵产物对软骨细胞的损伤。
The Fusarium oxysporum f. Sp. Strain 5-5-1 was a dominant Fusarium endophyte isolated from the wheat grains at the heading stage of KBD in Qinghai Province. In this experiment, the mycelium growth was measured by dry weight method. MTT (methylthiazolyldiphenyl -tetrazoliumbromide method was used to detect the chondrocyte survival rate. Toluidine blue staining and immunohistochemical staining were used to detect the expression of aggrecan and collagenⅡ in rabbit knee chondrocytes. The expression of reverse transcription-polymerase The expression of proteoglycan and type Ⅱ collagen m RNA in chondrocytes was detected by the method of reverse transcription-polymerase chain reaction (RT-PCR), the effect of selenium on the growth of 5-5-1 strain and the culture of this strain at different selenium levels Effect of Fermentation Products on Related Parameters of Rabbit Knee Chondrocytes. The results showed that the selenium concentration in the range of 0.05-0.5 mg / L promoted the growth of 5-5-1 strain and showed a concentration effect. The fermented product of this strain cultured under low concentration of selenium had a significant effect on chondrocyte proteoglycan And type II collagen synthesis and m RNA expression inhibition is relatively small. It was concluded that the appropriate concentration of selenium can promote the growth of Fusarium, but at the same time it can reduce the damage of chondrocytes by its fermentation products.