艾塞那肽对棕榈酸诱导成骨细胞凋亡的影响  

Effect of exenatide on the apoptosis of osteoblasts induced by palmitic acid

作　　者：柴利杰 辛莹[2] 李静怡[3] 王亭亭[2] 李珊[2] 郑丽丽[2] CHAI Li-jie;XIN Ying;LI Jing-yi;WANG Ting-ting;LI Shan;ZHENG Li-li(Department of Endocrinology and Metabolism,Fuwai Central China Cardiovascular Hospital,Zhengzhou 450046,China;Department of Endocrinology and Metabolism,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Plastic Surgery,The First Affiliated Hospital ofZhengzhou University,Zhengzhou 450052,China)

机构地区：[1]阜外华中心血管病医院内分泌科,郑州450046 [2]郑州大学第一附属医院内分泌科,郑州450052 [3]郑州大学第一附属医院整形外科,郑州450052

出　　处：《中华骨质疏松和骨矿盐疾病杂志》2018年第6期570-576,共7页Chinese Journal Of Osteoporosis And Bone Mineral Research

摘　　要：目的为探讨胰高血糖素样肽-1 (glucagon-like peptide-1,GLP-1)受体激动剂对骨保护作用的机制,应用GLP-1受体激动剂(艾塞那肽)干预高脂导致的小鼠成骨细胞凋亡实验,观察艾塞那肽在成骨细胞凋亡过程中的保护作用。方法体外应用含500μmol/L棕榈酸(palmitic acid,PA)完全培养基处理小鼠成骨细胞MC3T3-E1 14模拟体内高脂环境,分组如下:正常对照组(0μmol/L PA)、高脂组(500μmol/L PA)、高脂+艾塞那肽(1、10、100、1 000 nmol/L)浓度梯度组分别孵育成骨细胞24 h。根据实验结果选择艾塞那肽最佳浓度100 nmol/L进行后续实验,分正常对照组、艾塞那肽组、高脂+艾塞那肽组、高脂组,观察艾塞那肽对成骨细胞增殖和凋亡的影响。Cell Counting Kit-8试剂盒检测细胞增殖活性; Annexin V-FITC/PI凋亡检测试剂盒检测细胞凋亡率; RT-qPCR技术检测各组细胞内质网应激相关基因GRP78、CHOP、Caspase-12mRNA的表达。结果与正常对照组相比,高脂组细胞活性明显下降(OD值:2. 612±0. 106 vs. 2. 394±0. 088,P<0. 01);与高脂组相比,高脂+艾塞那肽1 000 nmol/L组、高脂+艾塞那肽100 nmol/L组、高脂+艾塞那肽10 nmol/L组细胞增殖活性明显升高(OD值:2. 394±0. 088、2. 547±0. 068、2. 579±0. 060、2. 496±0. 114,P<0. 05),高脂+艾塞那肽1 nmol/L组对细胞的增殖活力无明显影响(P> 0. 05)。选择艾塞那肽100 nmol/L为最佳浓度重复干预后发现,与高脂组相比,高脂+艾塞那肽100 nmol/L组的细胞增殖活力明显升高(OD值:2. 161±0. 203,2. 399±0. 193,P<0. 01);细胞凋亡率明显下降(19. 633%±3. 169%,5. 400%±2. 304%,P<0. 01); GRP78、CHOP mRNA的表达量明显下降(P <0. 01),Caspase-12 mRNA的表达量回升(P<0. 05)。结论 GLP-1受体激动剂艾塞那肽可能通过内质网应激途径,改善PA造成的小鼠成骨细胞增殖活性减低以及凋亡的发生。Objective To explore the mechanism of GLP-1 receptor agonists on bone protection, we used GLP-1 receptor agonist （Exenatide） to interfere with osteoblast apoptosis induced by high fat. Methods In vitro , we cultivated murine osteoblast MC3T3-E1 14 with complete medium containing 500 μmol/L palmitic acid （PA） mimicking high-fat environment in vivo for 24 hours. Six groups were divided normal control group, PA group, PA＋exenatide （1, 10, 100, 1 000 nmol/L） groups. According to the experimental results, the optimal concentration of 100 nmol/L was selected for follow-up experiments. Four groups were then divided： normal control group, exenatide group, PA＋exenatide group, PA group to observe the effects of exenatide on the proliferation and apoptosis of osteoblasts. Cell viability was detected by CCK-8 assay. Annexin V-FITC/PI apoptosis detection kit was used for detecting apoptosis rate. RT qPCR was used for detecting the GRP78, CHOP and Caspase-12 mRNA expression. Results Compared with the normal control group, the OD values of the cells in the PA group （ P 〈0.01） were decreased （OD value： 2.612±0.106, 2.394±0.088, P 〈0.01）. Compared with the PA group , the OD values of cells in the PA＋exenatide 1 000 nmol/L （ P 〈0.01）, PA＋exenatide 100 nmol/L （ P 〈0.01）, and PA＋exenatide 10 nmol/L （ P 〈0.05） were significantly increased （OD value： 2.394±0.088, 2.547±0.068, 2.579±0.060, 2.496±0.114, P 〈0.05）. The OD values of PA＋exenatide 1 nmol/L group had no significant effect on the osteoblasts proliferation （ P 〉0.05）. Compared with the PA group in the further experiment with the optimal concentration of 100 nmol/L , the OD values of PA＋ exenatide 100 nmol/L group were increased （OD value： 2.161±0.203, 2.399±0.193, P 〈0.01）; the apoptosis rate of PA＋exenatide 100 nmol/L group decreased significantly （19.633%±3.169%, 5.400%±2.304%, P 〈0.01）; the expression of GRP78 and CHOP mRNA in the endoplasmic reticulum stress-related genes w

关 键 词：成骨细胞 凋亡 艾塞那肽 内质网应激 

分 类 号：R681[医药卫生—骨科学]