论文部分内容阅读
目的探讨成骨细胞增殖分化中还原型谷胱甘肽的作用。方法清洁级3 d内新生Wistar大鼠3只,取颅骨剪碎成骨片,胰蛋白酶消化后培养成骨细胞,并采用苏木精-伊红染色、茜素红染色、矿化结节染色进行鉴定;MTT法检测高糖成骨细胞增殖情况;钙钴细胞化学染色法观察高糖成骨细胞分化情况。结果苏木精-伊红染色显示,成骨细胞呈梭形、三角形,胞浆丰富;茜素红染色显示,细胞多层重叠生长,部分区域积累灶状,形成钙结节;矿化结节染色显示,密集的钙化结节,细胞聚集。高糖组细胞增殖情况均低于同期正常对照组和谷胱甘肽组,差异有统计学意义(P<0.05);谷胱甘肽组细胞增殖情况高于同期高糖组,差异有统计学意义(P<0.05);谷胱甘肽组与同期正常对照组之间差异无统计学意义(P>0.05)。染色后显微镜下观察谷胱甘肽组存在大量染色细胞,胞浆为深蓝色,染色细胞数量明显多于高糖组,而正常对照组与谷胱甘肽组之间无明显差异。结论还原型谷胱甘肽可明显促进高糖环境下成骨细胞增殖、分化,预防糖尿病骨质疏松的发生。
Objective To investigate the role of reduced glutathione in the proliferation and differentiation of osteoblasts. Methods Three newborn Wistar rats were enrolled in this study. Three groups of newborn Wistar rats were scalded with skull and cut into osteoblasts. After trypsinization, osteoblasts were cultured and stained with hematoxylin-eosin, alizarin red and mineralized nodules The proliferation of hyperglycemic osteoblasts was detected by MTT assay and the differentiation of hyperglycemic osteoblasts was observed by calcium-cobalt cytochemical staining. Results The hematoxylin-eosin staining showed that the osteoblasts were spindle-shaped and triangular with abundant cytoplasm. Alizarin red staining showed that the cells were multi-layered and overlapped, and some areas formed focal distribution to form calcium nodules. The mineralized nodules Dyeing revealed dense clusters of calcified nodules and cells. The proliferation of high glucose group was lower than the normal control group and glutathione group, the difference was statistically significant (P <0.05); glutathione group cell proliferation was higher than the same period of high glucose group, the difference was statistically significant (P <0.05). There was no significant difference between glutathione group and normal control group (P> 0.05). After staining, there were a large number of stained cells in the glutathione group under the microscope. The cytoplasm was dark blue and the number of stained cells was significantly higher than that of the high glucose group. There was no significant difference between the normal control group and the glutathione group. Conclusion Reduced glutathione can significantly promote the proliferation and differentiation of osteoblasts under high glucose condition and prevent the occurrence of diabetic osteoporosis.