摘要
目的:探讨IL-31在促进骨质疏松过程中的作用及其机制。方法:选取本院2015年1月至2016年12月收治的100例骨质疏松患者为研究对象,并按性别、年龄匹配100例无骨质疏松的健康体检人群作为对照组,检测两组血清IL-31水平差异。构建C57BL/6J基因背景的IL-31敲除小鼠,将C57BL/6J小鼠(野生型小鼠)、IL-31敲除小鼠分别设置假手术(Sham)组和卵巢切除术(OVX)组各15只。手术8周后,检测小鼠血清中血钙、血磷水平和碱性磷酸酶水平(ALP);同时取小鼠的双侧股骨进行病理切片分析股骨组织形态结构、取脊柱进行骨密度仪检测脊柱骨密度变化。分离培养小鼠骨髓基质细胞(BMSCs)并检测体外增殖能力和TGF-β1、Smad4和p-Smad4蛋白表达。通过IL-31特异性shRNA转染小鼠成骨细胞MC3T3-E1以敲低IL-31的表达,并检测MC3T3-E1细胞增殖、凋亡和蛋白改变。结果:骨质疏松患者血清中IL-31水平显著高于对照组、重度骨质疏松患者的IL-31水平高于轻度骨质疏松患者(P<0. 05)。接受OVX手术的野生型小鼠、IL-31敲除小鼠血清中血钙和血磷显著高于假手术组,而ALP水平显著低于假手术组(P<0. 05),但接受OVX手术的IL-31敲除小鼠组血钙和血磷则显著低于接受OVX手术的野生型小鼠(P<0. 05)。股骨病理切片及脊柱骨密度结果显示,接受OVX手术的野生型小鼠、IL-31敲除小鼠均出现组织形态结构的破坏和骨密度下降,但IL-31敲除小鼠骨组织结构和骨密度优于野生型小鼠。接受OVX手术的IL-31敲除小鼠BMSCs增殖能力显著高于接受OVX手术的野生型小鼠,而TGF-β1、p-Smad4蛋白水平则显著高于接受OVX手术的野生型小鼠(P<0. 05)。MC3T3-E1细胞转染IL-31 shRNA后能明显反向激活TGF-β1、p-Smad4表达,促进MC3T3-E1细胞的增殖并抑制凋亡。结论:骨质疏松患者血清IL-31水平显著升高,IL-31可能是通过调控TGF-β1/Smad4信号通路抑制BMSCs增殖、促进成骨细胞的凋亡,从而
Objective:To investigate the role and mechanism of IL-31 in promoting osteoporosis.Methods:One hundred patients with osteoporosis and gender,aged matched healthy subjects without osteoporosis in our hospital from Jan 2015 to Dec 2016 were selected as study subjects,and the serum level of IL-31 in above two groups were compared.we also generaed IL-31 knockout(KO)mice in the C57BL/6J mice back ground.Wild type(WT)and IL-31 KO mice were received sham operation(Sham group)or ovariectomy(OVX group)for 15 mice in each group respectively.The serum level of calcium,phosphorus and alkaline phosphatase(ALP)were detected 8 weeks after operation.The bilateral femurs were collected after sacrifice to analyze the morphology and structure of the femur,and the spine were collected for detecting the bone density measured by bone density meter.The bone marrow stromal cells(BMSCs)were isolated and cultured in vitro.The proliferation ability of BMSCs,protein level of TGF-β1,Smad4 and p-Smad4 were detected.IL-31 shRNA was transfected into mouse osteoblast MC3T3-E1 cells to knockdown IL-31 and the proliferation,apoptosis and protein expression were detected.Results:The serum level of IL-31 in patients with osteoporosis were significantly higher than control group,and severe osteoporosis patients had significantly higher levels of IL-31 than mild osteoporosis patients(P<0.05).The serum level of calcium and phosphorus in OVX group of WT mice and IL-31 KO mice were significantly higher than the sham group(P<0.05),while the level of ALP was significantly lower than sham operation group(P<0.05),but the serum level of calcium and phosphorus in OVX group of IL-31 KO mice were significantly lower than WT mice(P<0.05).The pathological section of femur and spinal BMD showed that the bone tissue in WT mice and IL-31 KO received OVX were damaged and the bone density decreased significantly,but the damaged degree in IL-31 KO mice was significantly better than in WT mice.The proliferation ability BMSCs in OVX group of IL-31 KO mice was significant
作者
郝喆
杨文静
倪娟
HAO Zhe;YANG Wen-Jing;NI Juan(Department of Rheumatology and Immunology,Fourth Hospital of Wuhan,Wuhan 430033,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2019年第5期599-603,共5页
Chinese Journal of Immunology
