慢性肾脏病继发小鼠颌骨异常的形态学研究

›› 2020, Vol. 40 ›› Issue (5): 404-410.

慢性肾脏病继发小鼠颌骨异常的形态学研究

张耀元¹,王林¹,王华²,陈宏裕¹

1. 南京医科大学附属口腔医院
2. 南京医科大学口腔疾病研究江苏省重点实验室，南京医科大学附属口腔医院

收稿日期:2019-12-23 修回日期:2020-01-27 出版日期:2020-05-26 发布日期:2020-06-08
通讯作者: 王林 E-mail:lw603@njmu.edu.cn
基金资助:
江苏高校优势学科建设工程资助项目;科教强卫

Study of morphological feature and mechanism of maxillofacial bone abnormalities secondary to chronic kidney disease in mice

Received:2019-12-23 Revised:2020-01-27 Online:2020-05-26 Published:2020-06-08
Supported by:
A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions;the Jiangsu Provincial Key Medical Discipline

摘要/Abstract

摘要： ［摘要] 目的研究慢性肾脏病(chronic kidney disease, CKD)继发颌骨异常的形态学变化，并从细胞学角度对骨量变化进行探究。方法使用腺嘌呤饮食诱导建立CKD小鼠模型，每4周监测血清生化指标尿素氮(BUN)、肌酐(CREA)、钙(Ca)、磷(P)的变化，并于造模12周末行肾脏组织HE染色观察。造模12周末取小鼠下颌骨行Micro-CT和HE染色分析。采用酶联免疫吸附试验(ELISA)测定血清甲状旁腺激素(parathyroid hormone, PTH)水平。体外培养颌骨间充质干细胞(mesenchymal stem cells, MSCs)检测其增殖和成骨分化能力。诱导骨髓单核细胞(bone marrow macrophages, BMMs)成破骨细胞观察破骨分化能力。结果与正常对照组相比，CKD组的血清BUN、CREA、P和Ca水平升高(P＜0.05)，肾脏组织HE染色示肾小球数目减少，肾间质炎性细胞浸润和纤维化。Micro-CT示CKD组的下颌骨的骨密度降低，骨量下降(P＜0.05)；HE染色提示CKD组下颌骨类骨质增加，骨小梁形态不规则；ELISA示CKD组的血清PTH水平明显升高(P＜0.05)。体外实验显示，与正常对照组相比，CKD组MSCs增殖和早期成骨分化能力增强(P＜0.05)，但晚期形成矿化结节的能力有所下降(P＜0.05)；以及CKD组破骨细胞分化能力更强(P＜0.05)。结论 CKD继发甲状旁腺功能亢进可致MSCs高增殖，但晚期出现成骨矿化障碍，并且其破骨细胞分化能力增强，加速骨转化。颌骨表现为骨量下降，骨微结构改变。

关键词: 慢性肾脏病甲状旁腺激素骨微结构成骨细胞破骨细胞

Abstract: Abstract：Objective To investigate the morphological characteristics of maxillofacial bone abnormalities secondary to chronic kidney disease (CKD), meanwhile evaluating the biological function of mesenchymal stem cells (MSCs) and osteoclast precursors from the mouse model in vitro. Methods A mouse model of CKD was established by adenine diet induction. Monitor changes in body weight and serum biochemical indicators of blood urea nitrogen (BUN), creatinine (CREA), calcium (Ca), and phosphorus (P) every 4 weeks, and perform HE staining of kidney tissues at the end of the 12th week. The mandibles were analyzed by Micro-CT and HE staining. Enzyme-linked immunosorbent assay (ELISA) was used to determine the serum level of parathyroid hormone (PTH). MSCs were cultured in vitro to detect their proliferation and osteogenic differentiation ability. Bone marrow macrophage (BMMs) were induced into osteoclasts to observe the osteoclast differentiation. Results Compared with the normal control group, the body weights were decreased and serum levels of BUN, CREA, CA, P were increased in the CKD group(P<0.05). The HE staining of kidney tissues in the CKD group showed the number of glomerulus was decreased, renal interstitial inflammatory cell infiltration and fibrosis. Micro-CT analysis revealed that the bone density and mass of mandible were reduced in the CKD group (P<0.05); The HE staining of the CKD group showed an increase in mandibular osteoid and irregular trabecular shape. ELISA showed serum PTH levels were significantly increased in the CKD group (P<0.05). In vitro experiments showed that the proliferation and early osteogenic differentiation ability of MSCs of the CKD group were increased(P<0.05), but CKD osteoblasts mineralized slowly in the later period(P<0.05). The TRAP staining results showed osteoclast differentiation to be more active in the CKD group(P<0.05). Conclusions Hyperparathyroidism secondary to CKD can lead to osteogenesis and mineralization disorder of high-proliferation MSCs in the late stage，and contribute to the differentiation of osteoclast, and accelerate bone transformation. The maxillofacial bone may show a decrease in bone mass and changes in bone microstructure.

Key words: Chronic kidney disease parathyroid hormone bone microstructure Osteoblast Osteoclast

中图分类号:

R783.5

张耀元王林王华陈宏裕. 慢性肾脏病继发小鼠颌骨异常的形态学研究[J]. 口腔医学, 2020, 40(5): 404-410.

参考文献 20

[1]	Sagliker Y, Balal M, Sagliker Ozkaynak P, et al.Sagliker syndrome: uglifying human face appearance in late and severe secondary hyperparathyroidism in chronic renal failure[J].Seminars in nephrology, 2004, 24(5):449-455
[2]	马丹丹, 董静, 徐宝华.综合征病因的初步研究[J].中国医刊, 2012, 47(10):34-36
[3]	Pontes FSC, Lopes MA, de Souza LL, et al.Oral and maxillofacial manifestations of chronic kidney disease-mineral and bone disorder: a multicenter retrospective study[J].Oral surgery, oral medicine, oral pathology and oral radiology, 2018, 125(1):31-43
[4]	王庆婷，崔颖，李云飞等.调整腺嘌呤饮食磷含量制作慢性肾病高磷血症的小鼠模型[J].南京医科大学学报自然科学版, 2018, 38(07):956-961
[5]	Tani T, Orimo H, Shimizu A, et al.Development of a novel chronic kidney disease mouse model to evaluate the progression of hyperphosphatemia and associated mineral bone disease[J].Scientific reports, 2017, 7(1):2233-
[6]	Jia T, Olauson H, Lindberg K, et al.A novel model of adenine-induced tubulointerstitial nephropathy in mice 2013, 14: 116.[J].BMC nephrology, 2013, 14:116-
[7]	Williams MJ, Sugatani T, Agapova OA, et al.The activin receptor is stimulated in the skeleton,vasculature,heart,and kidney during chronic kidney disease[J].Kidney international, 2018, 93(1):147-158
[8]	杜海明, 张旗.缺失后小鼠下颌骨微结构增龄性变化的-评价[J].口腔颌面外科杂志, 2017, 27(01):13-18
[9]	Hsu PY, Tsai MT, Wang SP, et al.Cortical Bone Morphological and Trabecular Bone Microarchitectural Changes in the Mandible and Femoral Neck of Ovariectomized Rats[J].PloS one, 2016, 11(4):e0154367-
[10]	秦东泽，胡奥，郭威孝等.对不同时间和部位松质骨影响的-分析[J].口腔医学, 2017, 37(03):198-202
[11]	赵航, 盛青，苏俭生.小鼠颌骨间充质干细胞优化获取及鉴定 [J]. 2014, 24 (04):256-260.[J].口腔颌面外科杂志, 2014, 24 (04):256-260
[12]	李胜丹, 梁乙然, 刘一涵.衰老性骨质疏松微环境对颌骨骨髓间充质干细胞生物学功能的影响[J].中华老年口腔医学杂志, 2019, 17(04):198-203
[13]	Kim SH, Kim MO, Kim HJ, et al.Bortezomib prevents ovariectomy-induced osteoporosis in mice by inhibiting osteoclast differentiation[J].Journal of bone and mineral metabolism, 2018, 36(5):537-546
[14]	Ni LH, Tang RN, Lv LL, et al.A rat model of SHPT with bone abnormalities in CKD induced by adenine and a high phosphorus diet[J].Biochemical and biophysical research communications, 2018, 498(3):654-659
[15]	Isakov O, Rinella ES, Olchovsky D, et al.Missense mutation in the MEN1 gene discovered through whole exome sequencing co-segregates with familial hyperparathyroidism[J].Genetics research, 2013, 95(4):114-120
[16]	Jover Cerveró A, Bagán JV, Jiménez Soriano Y, et al.Dental management in renal failure: patients on dialysis[J].Medicina oral, patologia oral y cirugia bucal, 2008, 13(7):E419-426
[17]	Messier MD, Emde K, Stern L, et al.Radiographic periodontal bone loss in chronic kidney disease[J].Journal of periodontology, 2012, 83(5):602-611
[18]	Lee MM, Chu EY.El-Abbadi MM,et alCharacterization of mandibular bone in a mouse model of chronic kidney disease[J].J Periodontol, 2010, 81(2):300-309
[19]	江建青.尿毒症高转化骨病大鼠BM-MSC培养鉴定及增殖分化的信号转导机制 [D]. 天津医科大学, 2013.
[20]	Wu W, Qian LY, Chen XD, et al.A case of Sagliker syndrome and literature review[J].Zhong Nan Da Xue Xue Bao Yi Xue Ban, 2014, 39(10):1100-1104