摘要
目的:探讨红景天苷对诱导的成骨细胞在低氧环境中的保护作用及其机制。方法:组织块法分离培养小鼠成纤维细胞,转染重编程为多能诱导干细胞,进行Nanog、Oct-4、Rexl、Sox-2的QT-PCR检测。诱导性多潜能干细胞(IPS细胞)诱导分化为成骨细胞,进行茜素红染色鉴定。将诱导后的成骨细胞分为红景天苷预处理组、正常组和低氧组,用Annexin V-FITC/PI染色检测细胞凋亡率,Western blot检测HIF-1α和VEGF蛋白表达量。结果:多能诱导干细胞的内源性基因Nanog、Oct-4、Rexl、Sox-2的表达量均明显高于小鼠成纤维细胞(均P<0.05)。茜素红染色显示诱导后的成骨细胞染色阳性。红景天苷预处理组细胞凋亡率明显低于低氧组,差异有统计学意义(P<0.05)。红景天苷预处理组的HIF-1α和VEGF表达量高于低氧组(0.93±0.05 vs. 0.81±0.02,0.95±0.03 vs. 0.79±0.04;均P<0.05)。结论:红景天苷对诱导后成骨细胞在低氧环境中有保护作用,其机制可能是通过提高细胞HIF-1α和VEGF的表达来抑制成骨细胞的凋亡。
Objective:To investigate the protective effect of salidroside on induced osteoblasts in hypoxia environment and its mechanism. Methods:the mouse fibroblasts were separated and cultured by tissue block method. The transfected induced pluripotent stem cells were reprogrammed and the QT-PCR of Nanog, Oct-4, Rexl and Sox-2 were detected to the induced osteoblasts differentiated from the induced pluripotent stem cells and identified by alizarin red staining. The induced osteoblasts were divided into three groups:salidroside pre-conditioning group, normal oxygen group and hypoxia group. Annexin V-FITC/PI staining was used to detect the apoptosis rate, and Western-blot was used to detect VEGF, HIF-1 alpha. Results:The expression of endogenous gene Nanog, Oct-4, Rexl and Sox-2 of induced pluripotent stem cells was higher than that of mouse fibroblasts cells. The result is Nanog (783±42 vs. 308±23), Oct-4 (774±37 vs. 319±31), Rexl (742±28 vs. 382±31), Sox-2 (831±31 vs. 394±12). Alizarin red staining showed that the induced osteoblasts cells had positive staining results. The apoptotic rate of salidroside group was significantly lower than that of hypoxia group [(35.65±3.92)% vs.(95.20±1.03)%]. There was significant difference between the two groups (P<0.05). The expression of HIF-1a and VEGF in salidroside group was higher than that in hypoxic group (0.93±0.05 vs. 0.81±0.02, 0.95±0.03 vs. 0.79±0.04, P<0.05). Conclusion:Salidroside has protective effect on hypoxic environment of osteoblasts.
作者
沈新升
陈鸥
赖宪良
苏嘉
SHEN Xinsheng;CHEN Ou;LAIXianliang;SU Jia(Department of Orthopedic Surgery, Wenzhou Hospital of Integrated Traditional Chinese andWestern Medicine, Wenzhou 325000, China)
出处
《温州医科大学学报》
CAS
2019年第3期179-183,共5页
Journal of Wenzhou Medical University
基金
温州市公益性科技计划项目(Y20140594)
关键词
红景天苷
多能诱导干细胞
低氧
成骨细胞
凋亡
salidroside
induced pluripotent stem cells
hypoxia
osteoblast cells
apoptosis
