[1]徐仲翔,吴云刚,吴春雷.从骨髓基质干细胞活性的改变探讨激素性股骨头坏死的肾阳虚本质[J].中医正骨,2013,25(03):6-10.
　XU Zhong-xiang*,WU Yun-gang,WU Chun-lei..Study on the essence of Kidney-yang deficiency of steroid-induced avascular necrosis of femoral head from the activity change of marrow stroma cells[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2013,25(03):6-10.
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从骨髓基质干细胞活性的改变探讨激素性股骨头坏死的 肾阳虚本质()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第25卷
期数:
2013年03期
页码:
6-10
栏目:
股骨头坏死
出版日期:
2013-03-20

文章信息/Info

Title:
Study on the essence of Kidney-yang deficiency of steroid-induced avascular necrosis of femoral head from the activity change of marrow stroma cells
作者:
徐仲翔1; 吴云刚2; 吴春雷2
1.浙江省温州市第八人民医院,浙江 温州 325028;
2.温州医学院附属第一医院,浙江 温州 325000
Author(s):
XU Zhong-xiang*; WU Yun-gang; WU Chun-lei.
*The Eighth People's Hospital of Wenzhou City,Wenzhou 325028,Zhejiang,China
关键词:
股骨头坏死 骨髓细胞 氢化可的松 皮质醇 促肾上腺皮质激素 骨髓基质干细胞 肾阳虚
Keywords:
Femur head necrosis;  Bone marrow cells;  Hydrocortisone;  Cortisol;  Adrenocorticotropic hormone;  Marrow stroma cells;  Kidney-yang deficiency
摘要:
目的:探讨激素性股骨头坏死的肾阳虚本质。方法:在对6例创伤性股骨颈骨折患者(非激素性股骨头坏死组)和6例激素性股骨头坏死患者(激素性股骨头坏死组)行髋关节置换术前抽取其静脉血,采用放射免疫法测定血浆皮质醇和促肾上腺皮质激素的含量; 行髋关节置换术时分别取其股骨上段骨骺处液态的红骨髓,建立骨髓基质干细胞的体外培养体系,细胞传代培养并经体外成骨诱导后,采用MTT法检测成骨细胞的光密度值,进行碱性磷酸酶染色并检测成骨细胞的碱性磷酸酶含量,采用Von Kossa's染色法进行成骨细胞矿化结节计数,并在显微镜下观察成骨细胞形态。结果:①血浆皮质醇含量和促肾上腺皮质激素含量。激素性股骨头坏死组患者的血浆皮质醇含量和促肾上腺皮质激素含量[(328.167±27.499)μg·L-1,(11.450±2.649)ng·L-1]均低于非激素性股骨头坏死组[(380.500±25.618)μg·L-1,(15.667±3.751)ng·L-1],差异有统计学意义(t=3.411,P=0.007; t=2.249,P=0.048)。②成骨细胞形态。随着培养时间的延长,成骨细胞由贴壁→相互融合成单层→多层重叠生长。成骨细胞碱性磷酸酶染色呈强阳性,细胞质中出现黑色、灰褐色颗粒状或片状沉淀; 非激素性股骨头坏死组成骨细胞比激素性股骨头坏死组有不同程度的深染和范围扩大。Von Kossa's矿化结节染色显示,成骨细胞呈集落生长后开始形成矿化结节,细胞间出现致密的、圆形不透光团块,呈现片状的棕染; 非激素性股骨头坏死组成骨细胞矿化结节数比激素性股骨头坏死组明显增多。③成骨细胞光密度值、碱性磷酸酶含量及矿化结节数。激素性股骨头坏死组患者的成骨细胞光密度值、碱性磷酸酶含量、矿化结节数均低于非激素性股骨头坏死组[(0.174±0.035),(0.233±0.028),t=3.209,P=0.009;(1.685±0.323)U·L-1,(2.563±0.446)U·L-1,t=3.911,P=0.003;(18.000±3.578)个,(23.833±2.639)个,t=3.214,P=0.009]。结论:激素性股骨头坏死患者的下丘脑-垂体-肾上腺轴功能紊乱时,骨髓分化为成骨细胞的功能下降,成骨细胞的活性下降,这可能是激素性股骨头坏死患者肾阳虚在骨关节系统的具体体现,是大剂量激素导致肾阳虚并最终导致股骨头坏死的细胞学基础。
Abstract:
Objective:To explore the essence of Kidney-yang deficiency of steroid-induced avascular necrosis of femoral head(SANFH).Methods:Venous blood was collected from 6 patients with traumatic femoral neck fractures(non-SANFH group)and 6 SANFH patients(SANFH group)before hip replacement,and radioimmunoassay(RIA)was used to detect the contents of cortisol(CORT)and adrenocorticotropic hormone(ACTH)in the plasma.During hip replacement,the liquid red marrow was taken from proximal femoral osteoepiphysis to establish the culture system of marrow stroma cells(MSCs)in vitro.After the MSCs were serial subcultured and induced into osteoblasts in vitro,the optical density of osteoblasts was detected through MTT method,and alkaline phosphatase(ALP)staining was processed to measure ALP contents in osteoblasts,and the number of mineralized bone nodules in osteoblasts were counted through Von Kossa's staining method,then the morphous of osteoblasts was observed under microscope.Results:Contents of CORT and ACTH in the plasma of patients in SANFH group[(328.167±27.499)μg·L-1,(11.450±2.649)ng·L-1]were all lower than those in non-SANFH group[(380.500±25.618)μg·L-1,(15.667±3.751)ng·L-1],and there were statistical differences between the two groups(t=3.411,P=0.007; t=2.249,P=0.048).As time of culturing were prolonged,the osteoblasts adhered to the wall of the culture bottle,then they concentrated into single-layer,and they concentrated into multilayer in the end.After ALP staining,strong positive expressions of ALP were found in osteoblasts,and the black or fuscescent precipitate in granular or lamellar forms were found in the cytoplasm,and more obvious stain in more areas of osteoblasts were found in non-SANFH group compared with SANFH group.The results of Von Kossa's staining showed that the mineralized bone nodules began to appear after the osteoblasts gathered into colony; and the compact round lighttight clumps appeared between the cells,seen as a brown lamellar stain; and significantly more mineralized bone nodules were found in the osteoblasts of the non-SANFH group compared to SANFH group.The optical density,ALP contents and the number of mineralized bone nodules of osteoblasts in SANFH group were lower than those in non-SANFH group[(0.174±0.035),(0.233±0.028),t=3.209,P=0.009;(1.685±0.323)U·L-1,(2.563±0.446)U·L-1,t=3.911,P=0.003;(18.000±3.578),(23.833±2.639),t=3.214,P=0.009].Conclusion:The dysfunction of hypothalamus-pituitary-adrenal axis is followed by the inhibition of differentiation of marrow into osteoblasts and the inhibition of osteoblast activity,which may be the concrete reflection of Kidney-yang deficiency in the bone joint system of SANFH patients,and may be the cellular basis of the femoral head necrosis induced by Kidney-yang deficiency for the large dose of hormone.

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备注/Memo

备注/Memo:
基金项目:浙江省中医药科技计划项目(2008YB011),浙江省温州市鹿城区科技计划资助项目(S10102)
通讯作者:吴云刚 E-mail:zjwyg@126.com
更新日期/Last Update: 2013-03-20