中华地方病学杂志
中華地方病學雜誌
중화지방병학잡지
Chinese Journal of Endemiology
2014年
1期
37-40
,共4页
喻茂娟%王丽华%秦祥慧%金慰芳%高建军
喻茂娟%王麗華%秦祥慧%金慰芳%高建軍
유무연%왕려화%진상혜%금위방%고건군
氟%成骨细胞%骨保护素%核因子-κB受体活化因子配体
氟%成骨細胞%骨保護素%覈因子-κB受體活化因子配體
불%성골세포%골보호소%핵인자-κB수체활화인자배체
Fluorine%Osteoblasts%Osteoprotegerin%Receptor activator of NF-κB ligand
目的 探讨氟对体外培养大鼠成骨细胞中骨保护素(OPG)/核因子-κB受体活化因子配体(RANKL)/核因子-κB受体活化因子(RANK)信号传导通路的影响.方法 取新生24h内SD大鼠仔鼠,应用酶消化法分离大鼠颅骨成骨细胞,取第2代成骨细胞,加入不同浓度[0(对照)、1×10-3、1×104、1×10-5、1×10-6、1×10-7mol/L]的氟,培养24、48 h后,采用实时荧光定量PCR法检测成骨细胞RANKL、OPG mRNA表达;蛋白免疫印迹(Westem blot)法检测成骨细胞RANKL、OPG蛋白表达.结果 ①染氟24 h,成骨细胞RANKL、OPGmRNA表达组间比较差异有统计学意义(F值分别为30.95、22.62,P均＜0.01),其中1×10-5 mol/L组成骨细胞RANKL mRNA (5.99±0.39)和1×10-4、1×10-5、1×10-6 mol/L组OPG mRNA(3.52±0.09、4.81±0.15、3.68±0.04)的表达高于对照组(3.20±0.19、3.09±0.58,P均＜0.05),1×10-3 mol/L组RANKL mRNA (2.29±0.18)的表达低于对照组(P均＜0.05);染氟48 h,成骨细胞RANKL、OPG mRNA的表达组间比较差异有统计学意义(F值分别为26.62、5.72,P均＜0.01),其中1×10-5 mol/L组成骨细胞RANKL、OPG mRNA(6.67±0.49、5.05±0.51)的表达高于对照组(4.29±0.07、4.34±0.12,P均＜0.05),1×10-3 mol/L组OPG mRNA (3.63±0.49)的表达低于对照组(P＜0.05).②成骨细胞RANKL蛋白表达在染氟24、48 h后,组间比较差异无统计学意义(F值分别为0.07、0.49,P均＞0.05).成骨细胞OPG蛋白的表达,染氟24h后组间比较差异有统计学意义(F=3.26,P＜0.05),1×10-5 mol/L组(1.45±0.10)高于对照组(1.05±0.06,P＜0.05);染氟48 h,成骨细胞OPG蛋白表达组间比较差异无统计学意义(F=0.44,P＞ 0.05).结论 氟在低浓度时以成骨活动为主;但随着染氟时间和浓度的增加,促进破骨细胞的分化、成熟,以骨吸收为主.
目的 探討氟對體外培養大鼠成骨細胞中骨保護素(OPG)/覈因子-κB受體活化因子配體(RANKL)/覈因子-κB受體活化因子(RANK)信號傳導通路的影響.方法 取新生24h內SD大鼠仔鼠,應用酶消化法分離大鼠顱骨成骨細胞,取第2代成骨細胞,加入不同濃度[0(對照)、1×10-3、1×104、1×10-5、1×10-6、1×10-7mol/L]的氟,培養24、48 h後,採用實時熒光定量PCR法檢測成骨細胞RANKL、OPG mRNA錶達;蛋白免疫印跡(Westem blot)法檢測成骨細胞RANKL、OPG蛋白錶達.結果 ①染氟24 h,成骨細胞RANKL、OPGmRNA錶達組間比較差異有統計學意義(F值分彆為30.95、22.62,P均＜0.01),其中1×10-5 mol/L組成骨細胞RANKL mRNA (5.99±0.39)和1×10-4、1×10-5、1×10-6 mol/L組OPG mRNA(3.52±0.09、4.81±0.15、3.68±0.04)的錶達高于對照組(3.20±0.19、3.09±0.58,P均＜0.05),1×10-3 mol/L組RANKL mRNA (2.29±0.18)的錶達低于對照組(P均＜0.05);染氟48 h,成骨細胞RANKL、OPG mRNA的錶達組間比較差異有統計學意義(F值分彆為26.62、5.72,P均＜0.01),其中1×10-5 mol/L組成骨細胞RANKL、OPG mRNA(6.67±0.49、5.05±0.51)的錶達高于對照組(4.29±0.07、4.34±0.12,P均＜0.05),1×10-3 mol/L組OPG mRNA (3.63±0.49)的錶達低于對照組(P＜0.05).②成骨細胞RANKL蛋白錶達在染氟24、48 h後,組間比較差異無統計學意義(F值分彆為0.07、0.49,P均＞0.05).成骨細胞OPG蛋白的錶達,染氟24h後組間比較差異有統計學意義(F=3.26,P＜0.05),1×10-5 mol/L組(1.45±0.10)高于對照組(1.05±0.06,P＜0.05);染氟48 h,成骨細胞OPG蛋白錶達組間比較差異無統計學意義(F=0.44,P＞ 0.05).結論 氟在低濃度時以成骨活動為主;但隨著染氟時間和濃度的增加,促進破骨細胞的分化、成熟,以骨吸收為主.
목적 탐토불대체외배양대서성골세포중골보호소(OPG)/핵인자-κB수체활화인자배체(RANKL)/핵인자-κB수체활화인자(RANK)신호전도통로적영향.방법 취신생24h내SD대서자서,응용매소화법분리대서로골성골세포,취제2대성골세포,가입불동농도[0(대조)、1×10-3、1×104、1×10-5、1×10-6、1×10-7mol/L]적불,배양24、48 h후,채용실시형광정량PCR법검측성골세포RANKL、OPG mRNA표체;단백면역인적(Westem blot)법검측성골세포RANKL、OPG단백표체.결과 ①염불24 h,성골세포RANKL、OPGmRNA표체조간비교차이유통계학의의(F치분별위30.95、22.62,P균＜0.01),기중1×10-5 mol/L조성골세포RANKL mRNA (5.99±0.39)화1×10-4、1×10-5、1×10-6 mol/L조OPG mRNA(3.52±0.09、4.81±0.15、3.68±0.04)적표체고우대조조(3.20±0.19、3.09±0.58,P균＜0.05),1×10-3 mol/L조RANKL mRNA (2.29±0.18)적표체저우대조조(P균＜0.05);염불48 h,성골세포RANKL、OPG mRNA적표체조간비교차이유통계학의의(F치분별위26.62、5.72,P균＜0.01),기중1×10-5 mol/L조성골세포RANKL、OPG mRNA(6.67±0.49、5.05±0.51)적표체고우대조조(4.29±0.07、4.34±0.12,P균＜0.05),1×10-3 mol/L조OPG mRNA (3.63±0.49)적표체저우대조조(P＜0.05).②성골세포RANKL단백표체재염불24、48 h후,조간비교차이무통계학의의(F치분별위0.07、0.49,P균＞0.05).성골세포OPG단백적표체,염불24h후조간비교차이유통계학의의(F=3.26,P＜0.05),1×10-5 mol/L조(1.45±0.10)고우대조조(1.05±0.06,P＜0.05);염불48 h,성골세포OPG단백표체조간비교차이무통계학의의(F=0.44,P＞ 0.05).결론 불재저농도시이성골활동위주;단수착염불시간화농도적증가,촉진파골세포적분화、성숙,이골흡수위주.
Objective To study the influence of fluorine on signaling pathway of osteoprotegerin(OPG)/ receptor activator of NF-κB ligand(RANKL) in cultured rat osteoblasts.Methods Osteoblasts were isolated from skull of neonatal rats(＜ 24 hours) by enzyme digestion,and fluorine of different concentrations [0 (control),1 × 10-3,1 × 10-4,1 × 10-5,1 × l0-6 and 1 × 10-7 mol/L] were added into the culture medium of second generation of osteoblasts.The expressions of OPG and RANKL mRNA were determined using real-time PCR 24 and 48 hours after culturing.The expressions of OPG and RANKL protein were measured by Western blotting.Results ① After exposed to fluorine for 24 hours,the differences of RANKL and OPG mRNA expression had statistical significance between groups(F =30.95,22.62,all P ＜ 0.01),the expression of RANKL mRNA(5.99 ± 0.39) in the 1 × 10-5 mol/L group and the expressions of OPG mRNA(3.52 ± 0.09,4.81 ± 0.15,3.68 ± 0.04) in the 1 × 10-4,1 × 10-5 and 1 × 10-6 mol/L groups were higher than those of the control group(3.20 ± 0.19,3.09 ± 0.58,all P ＜ 0.05),but in the 1 × 10-3 mol/L group,RANKL mRNA(2.29 ± 0.18) was lower than that of the control group(P ＜ 0.05).After exposed to fluorine for 48 hours,the differences of RANKL and OPG mRNA expression had statistical significance between groups(F =26.62,5.72,all P ＜ 0.01),the expressions of RANKL and OPG mRNA(6.67 ± 0.49 and 5.05 ± 0.51) in the 1 × 10-5 mol/L group were higher than those of the control group(4.29 ± 0.07 and 4.34 ± 0.12,all P ＜ 0.05),and in the 1 × 10-3 mol/L group the expression of OPG mRNA(3.63 ± 0.49) was lower than that of the control group(P ＜ 0.05).② The expression of RANKL protein was not statistically significant between 24 hours and 48 hours groups (F =0.07,0.49,all P ＞ 0.05) ; the differences of OPG protein expression had statistical significance between groups(F =3.26,P ＜ 0.05),the expression of OPG protein in the 1 × 10-5 mol/L group(1.45 ± 0.10) was higher than that of the control group(1.05 ± 0.06,P ＜ 0.05) at the 24 hours.After 48 hours,the expression of OPG protein was not statistically significant(F =0.44,P ＞ 0.05).Conclusions At lower fluorine concentrations,bone formation is the main activity.But when fluorine concentration increased and time prolonged,the osteoclast differentiation and maturation are promoted,and the bone resorption is the main thing.
