IL-38通过调控PI3K/Akt/GSK3β/NFATc1信号通路抑制骨质疏松的机制研究  

IL-38 inhibits osteoporosis via regulating PI3K/Akt/GSK3β/NFATc1 signaling pathway

作　　者：刘珍星 张山锋[1] 杨钟华[1] LIU Zhen-Xing;ZHANG Shan-Feng;YANG Zhong-Hua(Department of Orthopedics, Wuhan General Hospital ofTongji Medical College,Huazhong University of Science and Technology,Wuhan 430034,Chin)

机构地区：[1]华中科技大学同济医学院附属武汉普爱医院骨科,武汉430034

出　　处：《中国免疫学杂志》2018年第2期251-255,共5页Chinese Journal of Immunology

摘　　要：目的:探讨IL-38抑制骨质疏松的作用并研究其分子机制。方法:共纳入2014年6月~2016年12月我院收治的138例骨质疏松患者作为研究对象。另选取120例同期在我院骨科进行骨折手术的无骨质疏松患者作为对照。采用ELISA法检测实验对象血清IL-38水平。构建IL-38-C57BL/6J转基因小鼠,建立骨质疏松小鼠模型,将野生型、IL-38转基因小鼠分别设置为假手术组(Sham组)与卵巢切除组(Ovariectomy,OVX组)。术后8周取小鼠血清,检测碱性磷酸酶(ALP)、血钙及血磷水平。另取小鼠的脊柱与双侧股骨,通过病理切片分析股骨组织形态结构,用骨密度仪检测脊柱骨密度变化。将各组小鼠的骨髓基质细胞(BMSCs)进行分离并检测其体外增殖能力,Western blot检测各组BMSCs的PI3K、Akt、GSK3β与NFATc1的磷酸化水平。小鼠成骨细胞MC3T3-E1转染IL-38后,Western blot检测PI3K、Akt、GSK3β与NFATc1磷酸化水平的变化,流式细胞术检测IL-38对细胞凋亡的影响。结果:骨质疏松组患者的血清IL-38水平显著低于对照组(P<0.05)。野生型与IL-38转基因OVX小鼠的血钙、血磷水平均显著高于Sham组(P<0.05),而ALP水平显著低于Sham组(P<0.05)。另外,IL-38转基因OVX小鼠的血钙和血磷水平均显著低于野生型OVX小鼠(P<0.05)。股骨病理切片及脊柱骨密度分析显示,野生型与IL-38转基因OVX小鼠均出现骨组织形态结构破坏和骨密度下降,并且IL-38转基因OVX小鼠的骨组织形态结构破坏和骨密度下降情况均较野生型OVX小鼠显著减轻(P<0.05)。IL-38转基因OVX小鼠BMSCs的体外增殖能力显著高于野生型OVX组(P<0.05)。IL-38转基因OVX小鼠BMSCs的PI3K、Akt与NFATc1磷酸化水平均显著低于野生型OVX组(P<0.05),GSK3β磷酸化水平显著高于野生型OVX组(P<0.05)。MC3T3-E1细胞转染IL-38后PI3K、Akt与NFATc1的磷酸化水平均显著降低(P<0.05),GSK3β的磷酸化水平显著升高(P<0.05)。流式细胞检测显示转染IL-38后MC3T3-EObjective： To investigate the role and mechanism of IL-38 in inhibiting osteoporosis. Methods： A total of 138 cases of patients with osteoporosis in our hospital from June 2014 to December 2016 were recruited. Another 120 cases of fracture surgery patients without osteoporosis were selected as control. Serum levels of IL-38 in different groups were determined using a commercially available sandwich ELISA（ Enzyme-Linked Immuno Sorbent Assay）. Construction of IL-38-C57 BL/6 J transgenic mice,the wild type and IL-38 transgenic mice were set to sham operation group（ Sham）,operation group（ ovariectomy,group OVX） respectively. After 8 weeks of the operation,the serum level of alkaline phosphatase（ ALP）,calcium and phosphorus were detected. The bilateral femur and spine of mice were collected after sacrifice,the morphology and structure of the femur were analyzed,and the bone density was measured by bone density meter. The bone marrow stromal cells（ BMSCs） were isolated and the invitro proliferation ability of BMSCs were measured. Western blot were used to detect the phosphorylation level of PI3 K,Akt,GSK3β and NFATc1 in BMSCs. After transfection of IL-38 into mouse osteoblast MC3 T3-E1 cell,the phosphorylation level of PI3 K,Akt,GSK3β and NFATc1 were detected by Western blot. Apoptosis of MC3 T3-E1 cells were detected by flow cytometry. Results： The serum level of IL-38 in patients with osteoporosis were significant lower than control group（ P〈0. 05）. The serum level of estrogen,calcium and phosphorus in OVX group of wild type and IL-38 transgenic mice were significant lower than the sham operation group（ P〈0. 05）,while the level of ALP was significant higher than sham operation group（ P〈0. 05）,but the serum level of calcium and phosphorus in OVX group of IL-38 transgenic mice were significant higher than wild type mice（ P〈0. 05）. The pathological section of femur and spine BMD showed that the bone tissue in wild type mice and IL-38 transgenic mice in OVX group were d

关 键 词：IL-38 PI3K AKT GSK3Β NFATc1 骨质疏松 

分 类 号：R-34[医药卫生]