论文部分内容阅读
目的:观察急性高糖环境对大鼠骨形成的影响。方法:将原代新生SD大鼠成骨细胞分为对照组(葡萄糖浓度为5.5 mmol/L)和高糖组(葡萄糖浓度为22.0 mmol/L),高糖组分别于作用后24、48、72 h收集细胞。向成年SD大鼠颈内静脉持续灌注生理盐水(对照组)和50%葡萄糖(高糖组),持续灌注24、48、72 h后处死,并取胫腓骨骨组织。分别提取成骨细胞和胫腓骨骨组织中骨钙素(OC)、Ⅰ型胶原(COL1)、Runt相关转录因子2(RUNX2)、碱性磷酸酶(ALP)、c-JUN和胰岛素样生长因子1(IGF-1)的mRNA及相应蛋白,并用实时荧光定量PCR和蛋白印迹法检测骨形成相关基因的表达。结果:急性高糖环境下,相较对照组,大鼠成骨细胞ALP、COL1、RUNX2、IGF-1、OC的mRNA表达在高糖培养48 h升高(P<0.05);成骨细胞经高糖处理后,P38MAPK磷酸化水平逐渐增加,在高糖培养48 h时磷酸化水平达最高。在不同高糖处理时间组SD大鼠胫腓骨骨组织中,OC、ALP、COL1、RUNX2、IGF-1、c-JUN的mRNA表达有差异,P38MAPK磷酸化水平在高糖培养48 h达最高。结论:骨形成过程中,成骨细胞可能在急性高糖环境下出现增殖分化增加,并通过MAPK通路进行调控。
Objective: To observe the effect of acute hyperglycemic environment on bone formation in rats. Methods: Primary cultured neonatal SD rat osteoblasts were divided into control group (glucose concentration 5.5 mmol / L) and high glucose group (glucose concentration 22.0 mmol / L) 72 h to collect cells. Adult SD rats were injected intraperitoneally with normal saline (control group) and 50% glucose (high glucose group). The rats were sacrificed 24, 48, and 72 hours after operation, and the tibia and fibula were removed. The expressions of osteocalcin (OC), collagen type Ⅰ (COL1), Runt-associated transcription factor 2 (RUNX2), alkaline phosphatase (ALP), c-JUN and insulin-like growth factor 1 (IGF-1) mRNA and protein, and the expression of bone formation-related genes was detected by real-time fluorescence quantitative PCR and Western blotting. Results: Compared with the control group, the mRNA expression of ALP, COL1, RUNX2, IGF-1 and OC in rat osteoblasts increased significantly (P <0.05) Phosphorylation level of P38MAPK gradually increased after high glucose treatment, and phosphorylation reached the highest at 48 h after high glucose. The mRNA expressions of OC, ALP, COL1, RUNX2, IGF-1 and c-JUN in tibia and fibula of SD rats were significantly different at different time points of high glucose treatment. The phosphorylation of P38MAPK was highest at 48 h after high glucose. Conclusion: During osteogenesis, osteoblasts may proliferate and differentiate under acute hyperglycemia and regulate through MAPK pathway.