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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:

第25卷

期數(shù):

2013年03期

頁碼:

6-10

欄目:

股骨頭壞死

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; 吳云剛²; 吳春雷²

1.浙江省溫州市第八人民醫(yī)院,浙江 溫州 325028;
2.溫州醫(yī)學(xué)院附屬第一醫(yī)院,浙江 溫州 325000

Author(s):

XU Zhong-xiang^*; WU Yun-gang; WU Chun-lei.

^*The Eighth People's Hospital of Wenzhou City,Wenzhou 325028,Zhejiang,China

關(guān)鍵詞:

股骨頭壞死 骨髓細胞 氫化可的松 皮質(zhì)醇 促腎上腺皮質(zhì)激素 骨髓基質(zhì)干細胞 腎陽虛

Keywords:

Femur head necrosis;  Bone marrow cells;  Hydrocortisone;  Cortisol;  Adrenocorticotropic hormone;  Marrow stroma cells;  Kidney-yang deficiency

摘要:

目的:探討激素性股骨頭壞死的腎陽虛本質(zhì)。方法:在對6例創(chuàng)傷性股骨頸骨折患者(非激素性股骨頭壞死組)和6例激素性股骨頭壞死患者(激素性股骨頭壞死組)行髖關(guān)節(jié)置換術(shù)前抽取其靜脈血,采用放射免疫法測定血漿皮質(zhì)醇和促腎上腺皮質(zhì)激素的含量; 行髖關(guān)節(jié)置換術(shù)時分別取其股骨上段骨骺處液態(tài)的紅骨髓,建立骨髓基質(zhì)干細胞的體外培養(yǎng)體系,細胞傳代培養(yǎng)并經(jīng)體外成骨誘導(dǎo)后,采用MTT法檢測成骨細胞的光密度值,進行堿性磷酸酶染色并檢測成骨細胞的堿性磷酸酶含量,采用Von Kossa's染色法進行成骨細胞礦化結(jié)節(jié)計數(shù),并在顯微鏡下觀察成骨細胞形態(tài)。結(jié)果:①血漿皮質(zhì)醇含量和促腎上腺皮質(zhì)激素含量。激素性股骨頭壞死組患者的血漿皮質(zhì)醇含量和促腎上腺皮質(zhì)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頭壞死組[(380.500±25.618)μg·L^-1,(15.667±3.751)ng·L^-1],差異有統(tǒng)計學(xué)意義(t=3.411,P=0.007; t=2.249,P=0.048)。②成骨細胞形態(tài)。隨著培養(yǎng)時間的延長,成骨細胞由貼壁→相互融合成單層→多層重疊生長。成骨細胞堿性磷酸酶染色呈強陽性,細胞質(zhì)中出現(xiàn)黑色、灰褐色顆粒狀或片狀沉淀; 非激素性股骨頭壞死組成骨細胞比激素性股骨頭壞死組有不同程度的深染和范圍擴大。Von Kossa's礦化結(jié)節(jié)染色顯示,成骨細胞呈集落生長后開始形成礦化結(jié)節(jié),細胞間出現(xiàn)致密的、圓形不透光團塊,呈現(xiàn)片狀的棕染; 非激素性股骨頭壞死組成骨細胞礦化結(jié)節(jié)數(shù)比激素性股骨頭壞死組明顯增多。③成骨細胞光密度值、堿性磷酸酶含量及礦化結(jié)節(jié)數(shù)。激素性股骨頭壞死組患者的成骨細胞光密度值、堿性磷酸酶含量、礦化結(jié)節(jié)數(shù)均低于非激素性股骨頭壞死組[(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)個,(23.833±2.639)個,t=3.214,P=0.009]。結(jié)論:激素性股骨頭壞死患者的下丘腦-垂體-腎上腺軸功能紊亂時,骨髓分化為成骨細胞的功能下降,成骨細胞的活性下降,這可能是激素性股骨頭壞死患者腎陽虛在骨關(guān)節(jié)系統(tǒng)的具體體現(xiàn),是大劑量激素導(dǎo)致腎陽虛并最終導(dǎo)致股骨頭壞死的細胞學(xué)基礎(chǔ)。

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.

備注/Memo

備注/Memo:

基金項目:浙江省中醫(yī)藥科技計劃項目(2008YB011),浙江省溫州市鹿城區(qū)科技計劃資助項目(S10102)
通訊作者:吳云剛 E-mail:[email protected]

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更新日期/Last Update: 2013-03-20