強骨寶對去卵巢大鼠骨量指標與形態(tài)計量參數(shù)的比較觀察

1、強骨寶對去卵巢大鼠骨量指標與形態(tài)計量參數(shù)的比較觀察         08-05-24 16:39:00     編輯：studa20             作者：王永東，周亞民，吳鐵，劉曉青【關(guān)鍵詞】  骨質(zhì)疏松癥；,大鼠；,骨組織形態(tài)計量學(xué)；,黃芪；,司坦唑醇 摘要：目的觀察強骨寶復(fù)方制劑干預(yù)治療去卵巢骨質(zhì)疏松癥模型大鼠脛骨鈣、鎂、磷

2、及羥脯氨酸的含量和骨形態(tài)計量參數(shù)的變化，并討論兩者變化的相互關(guān)系。方法取32只Wistar雌性大鼠隨機分為4組，按要求行去勢手術(shù)，然后分別灌胃給藥，8周后處死，取大鼠右脛骨烘干測定鈣、鎂、磷及羥脯氨酸；取左脛骨上段松質(zhì)骨和中段皮質(zhì)骨硬組織包埋、切片，圖像分析松質(zhì)骨、皮質(zhì)骨形態(tài)計量參數(shù)。結(jié)果強骨寶可使去卵巢大鼠脛骨干重，羥脯氨酸、鈣、鎂與骨干重的比值等骨質(zhì)量指標明顯提高；去卵巢大鼠脛骨松質(zhì)骨靜態(tài)參數(shù)和動態(tài)參數(shù)顯著增加，吸收參數(shù)顯著降低；皮質(zhì)骨靜態(tài)及動態(tài)參數(shù)均無顯著變化，內(nèi)膜面骨吸收參數(shù)降低。結(jié)論強骨寶對去卵巢大鼠骨質(zhì)疏松癥的作用，從骨形態(tài)計量參數(shù)變化上看能促進骨形成，抑制骨吸收，增加松質(zhì)骨的密度

3、；從骨質(zhì)量指標變化來看能促進骨羥脯氨酸等骨膠原有機質(zhì)的分泌合成，并增加成骨細胞對鈣、鎂、磷等無機礦物質(zhì)的吸收和沉積，從而促進新骨的形成，骨質(zhì)量的增加。因此，骨形態(tài)計量參數(shù)與骨質(zhì)量指標兩者的變化是統(tǒng)一的。關(guān)鍵詞：骨質(zhì)疏松癥；  大鼠；  骨組織形態(tài)計量學(xué)；  黃芪；  司坦唑醇The Comparatiue Effect of JGB on the Bone Histomorphometry and Bone Quality Parameter in Ovariectomized RatsAbstract：ObjectiveTo investigate t

4、he changes of the compound of  Astragalus membranaceus and Stanozolol (JGB) on the bone histomorphometry and bone quality parameter such as Calcium,Magnesium,Phosphorus,hydroxyproline and bone weight in ovariectomized(OVX) osteoprosis rats, and analyse the mutuality between the changes.Methods3

5、2 Wistar female rats, randomly divided into 4 groups were shamly operated or ovariectomized ,then all were fed with medicine for two months and then killed.The right tibiae were roasted and weighed to be dissolved by muriatic acid for testing Calcium,Magnesium,Phosphorus, and hydroxyproline. The lef

6、t proximal and middle tibias were trimed and processed undercalcified sections for cancellous and cortical bone histomorphometric analysis.ResultsThe bone quality parameter such as Calcium,Magnesium,Phosphorus,hydroxyproline and bone weight obviously increased, the static and dynamic parameters of t

7、he cancellous bone histomorphometry similarly significantly increased,but the resorption parameters decreased markedly; meantime,the static and dynamic parameters of the cortical bone histomorphometry both had no significant change, while the bone resorption parameter in the endocortical region decr

8、eased obviously.ConclusionThe cancellous bone density of OVX rats can be evidently improved by increasing bone formation and inhibiting the endocortical bone resorption.The bone quality also be markedly improved by enhancing hydroxyproline secretion and osteoblast resorption to Calcium, Magnesium an

9、d Phosphorus and leading to new bone reconstruction .Therefore, the changes of the bone histomorphometry and bone quality parameter in ovariectomized rats have the same consistency.Key words：Osteoporosis；  Rats；  Bone histomorphometry；  Astragalus membranaceus；  Stanozolol  

10、   骨質(zhì)疏松癥常表現(xiàn)為機體及骨組織生理性退行性變化，骨量減少，骨的結(jié)構(gòu)改變和骨折危險度增加1。本實驗強骨寶由中藥黃芪和司坦唑醇組成復(fù)方，以去卵巢大鼠為骨質(zhì)疏松癥的動物模型，在強骨寶的作用下，觀察大鼠脛骨骨形態(tài)計量學(xué)參數(shù)和鈣、鎂、磷、羥脯氨酸等骨質(zhì)量指標的變化，并討論其兩者變化的相互關(guān)系。1  材料與方法1.1  動物及分組3月齡Wistar普通級雌性大鼠32只（廣東醫(yī)學(xué)院實驗動物中心提供），體重(268±22)g , 隨機分為4組：正常對照組（Sham）、去卵巢對照組（OVX）、強骨寶組(JGB)、己烯雌酚組(DES)。除正常對照組外，其它3

11、組均行去勢手術(shù)。術(shù)后1 d，對照組、強骨寶組、己烯雌酚組分別以生理鹽水、強骨寶（生藥含量為2 g/ml）、己烯雌酚（0.5 mg/ml）按大鼠體重0.5 ml/100 g灌胃給藥8周時間后處死。1.2  藥品及試劑己烯雌酚，廣東華南藥業(yè)集團，批號050403；司坦唑醇，廣西百會藥業(yè)集團，批號050512；強骨寶，自制復(fù)方，主要成份為中藥黃芪提取物及司坦唑醇。1.3  骨組織形態(tài)計量學(xué)測量及參數(shù)2全部大鼠在處死前第14，13天和第4，3天分別給其皮下注射鹽酸四環(huán)素(25 mgkg-1)和鈣黃綠素 ( 5mgkg-1)，以在骨表面形成黃色和綠色兩層雙熒光標記，動態(tài)觀察兩次注射期

12、間骨形成的情況。大鼠給藥8周后測定體重，處死解剖取左脛骨，脛骨上段按硬組織切片法進行不脫鈣骨切片包埋、染色處理。所制切片在距骨骺線1 mm處至遠端3 mm范圍內(nèi)進行松質(zhì)骨形態(tài)計量學(xué)參數(shù)測量；脛骨中段皮質(zhì)骨不脫鈣骨磨片處理，按皮質(zhì)骨骨形態(tài)計量學(xué)方法測量和參數(shù)計算骨形態(tài)計量學(xué)靜態(tài)參數(shù)包括：松質(zhì)骨骨小梁面積百分率（%Tb.Ar），松質(zhì)骨骨小梁分離度(Tb.Sp)，皮質(zhì)骨組織面積百分數(shù)（%Ct.Ar）；吸收參數(shù)：單位骨小梁面積破骨細胞數(shù)量（Oc.N）；動態(tài)參數(shù)包括：松質(zhì)骨骨小梁熒光標志周長百分率（%L.Pm ），松質(zhì)骨骨小梁礦化沉積率(MAR)；皮質(zhì)骨膜熒光標記周長百分數(shù)（P-%L.Pm）、皮質(zhì)骨骨膜骨礦化沉積率（P-MAR）。1.4  骨礦質(zhì)元素及羥脯氨酸（HOP）測定取右脛骨，除凈附著骨上的肌肉和軟組織，烘干至恒重，稱