海南医学
海南醫學
해남의학
HAINAN MEDICAL JOURNAL
2014年
22期
3286-3289
,共4页
傅坤发%陆冰%刘晔%胡健%孔繁荣
傅坤髮%陸冰%劉曄%鬍健%孔繁榮
부곤발%륙빙%류엽%호건%공번영
辛伐他汀%成骨细胞%经典Wnt信号通路%骨质疏松
辛伐他汀%成骨細胞%經典Wnt信號通路%骨質疏鬆
신벌타정%성골세포%경전Wnt신호통로%골질소송
Simvastatin%Osteoblasts%Canonical Wnt signaling pathway%Osteoporosis
目的：观察不同浓度、不同时间辛伐他汀对体外培养的大鼠成骨细胞经典Wnt通路Wnt3a、LRP5和β-catenin的mRNA表达的影响，探讨辛伐他汀抗骨质疏松的可能机制。方法用酶消化法培养大鼠颅骨成骨细胞，采用倒置相差显微镜及碱性磷酸酶染色和茜素红染色观察和鉴定成骨细胞。用浓度分别为10-8 mol/L、10-7 mol/L、10-6 mol/L和10-5 mol/L的辛伐他汀干预大鼠成骨细胞24 h、48 h和72 h，并设无辛伐他汀为对照组，采用RT-PCR检测经典Wnt信号通路Wnt3a、LRP5和β-catenin的mRNA表达情况。结果镜下观察及染色结果均显示具有成骨细胞典型特征。RT-PCR结果显示：10-5 mol/L辛伐他汀组干预24 h后较对照组Wnt3a [(2.09±0.12) vs (1.82±0.09)]、LRP5[(1.42±0.07) vs (1.29±0.07)]和β-catenin [(1.10±0.06) vs (0.95±0.06)]的mRNA表达显著增加；干预48 h和72 h后各组Wnt3a (F=37.78和96.52，P=0.000)、LRP5(F=10.32，P=0.001和F=22.02，P=0.000）的mRNA表达随着辛伐他汀浓度的增大而显著增加，β-catenin的mRNA表达较对照组显著增加(F=6.67和21.90，P<0.05)；10-8~10-5 mol/L辛伐他汀各组中分别干预24 h、48 h和72 h，随着干预时间延长Wnt3a和LRP5的mRNA表达均增加；干预48 h与24 h比较，β-catenin的mRNA表达差异无统计学意义[(1.08±0.09) vs (1.02±0.06)，P>0.05]，而干预72 h较24 h显著增加[(1.23±0.10) vs (1.02±0.06)，P<0.05]，且辛伐他汀浓度为10-6 mol/L和10-5 mol/L干预72 h较48 h显著增加[(1.32±0.07) vs (1.11±0.05)和(1.47±0.08) vs (1.19±0.04)，P<0.05]。结论辛伐他汀可促进体外培养大鼠成骨细胞经典Wnt信号通路Wnt3a、LRP5和β-catenin的mRNA表达。
目的：觀察不同濃度、不同時間辛伐他汀對體外培養的大鼠成骨細胞經典Wnt通路Wnt3a、LRP5和β-catenin的mRNA錶達的影響，探討辛伐他汀抗骨質疏鬆的可能機製。方法用酶消化法培養大鼠顱骨成骨細胞，採用倒置相差顯微鏡及堿性燐痠酶染色和茜素紅染色觀察和鑒定成骨細胞。用濃度分彆為10-8 mol/L、10-7 mol/L、10-6 mol/L和10-5 mol/L的辛伐他汀榦預大鼠成骨細胞24 h、48 h和72 h，併設無辛伐他汀為對照組，採用RT-PCR檢測經典Wnt信號通路Wnt3a、LRP5和β-catenin的mRNA錶達情況。結果鏡下觀察及染色結果均顯示具有成骨細胞典型特徵。RT-PCR結果顯示：10-5 mol/L辛伐他汀組榦預24 h後較對照組Wnt3a [(2.09±0.12) vs (1.82±0.09)]、LRP5[(1.42±0.07) vs (1.29±0.07)]和β-catenin [(1.10±0.06) vs (0.95±0.06)]的mRNA錶達顯著增加；榦預48 h和72 h後各組Wnt3a (F=37.78和96.52，P=0.000)、LRP5(F=10.32，P=0.001和F=22.02，P=0.000）的mRNA錶達隨著辛伐他汀濃度的增大而顯著增加，β-catenin的mRNA錶達較對照組顯著增加(F=6.67和21.90，P<0.05)；10-8~10-5 mol/L辛伐他汀各組中分彆榦預24 h、48 h和72 h，隨著榦預時間延長Wnt3a和LRP5的mRNA錶達均增加；榦預48 h與24 h比較，β-catenin的mRNA錶達差異無統計學意義[(1.08±0.09) vs (1.02±0.06)，P>0.05]，而榦預72 h較24 h顯著增加[(1.23±0.10) vs (1.02±0.06)，P<0.05]，且辛伐他汀濃度為10-6 mol/L和10-5 mol/L榦預72 h較48 h顯著增加[(1.32±0.07) vs (1.11±0.05)和(1.47±0.08) vs (1.19±0.04)，P<0.05]。結論辛伐他汀可促進體外培養大鼠成骨細胞經典Wnt信號通路Wnt3a、LRP5和β-catenin的mRNA錶達。
목적：관찰불동농도、불동시간신벌타정대체외배양적대서성골세포경전Wnt통로Wnt3a、LRP5화β-catenin적mRNA표체적영향，탐토신벌타정항골질소송적가능궤제。방법용매소화법배양대서로골성골세포，채용도치상차현미경급감성린산매염색화천소홍염색관찰화감정성골세포。용농도분별위10-8 mol/L、10-7 mol/L、10-6 mol/L화10-5 mol/L적신벌타정간예대서성골세포24 h、48 h화72 h，병설무신벌타정위대조조，채용RT-PCR검측경전Wnt신호통로Wnt3a、LRP5화β-catenin적mRNA표체정황。결과경하관찰급염색결과균현시구유성골세포전형특정。RT-PCR결과현시：10-5 mol/L신벌타정조간예24 h후교대조조Wnt3a [(2.09±0.12) vs (1.82±0.09)]、LRP5[(1.42±0.07) vs (1.29±0.07)]화β-catenin [(1.10±0.06) vs (0.95±0.06)]적mRNA표체현저증가；간예48 h화72 h후각조Wnt3a (F=37.78화96.52，P=0.000)、LRP5(F=10.32，P=0.001화F=22.02，P=0.000）적mRNA표체수착신벌타정농도적증대이현저증가，β-catenin적mRNA표체교대조조현저증가(F=6.67화21.90，P<0.05)；10-8~10-5 mol/L신벌타정각조중분별간예24 h、48 h화72 h，수착간예시간연장Wnt3a화LRP5적mRNA표체균증가；간예48 h여24 h비교，β-catenin적mRNA표체차이무통계학의의[(1.08±0.09) vs (1.02±0.06)，P>0.05]，이간예72 h교24 h현저증가[(1.23±0.10) vs (1.02±0.06)，P<0.05]，차신벌타정농도위10-6 mol/L화10-5 mol/L간예72 h교48 h현저증가[(1.32±0.07) vs (1.11±0.05)화(1.47±0.08) vs (1.19±0.04)，P<0.05]。결론신벌타정가촉진체외배양대서성골세포경전Wnt신호통로Wnt3a、LRP5화β-catenin적mRNA표체。
Objective To investigate the effect of different concentrations, multi-treatment course of simvas-tatin on the mRNA expression of Wnt3a, LRP5 and β-catenin in osteoblasts of rats and the potential mechanism. Methods Osteoblasts were obtained from calvaria. Morphology of osteoblasts was observed under phase contrast mi-croscope after ALP staining and Alizarin red staining. Rat osteoblasts were treated by different concentrations of simvas-tatin (10-8 mol/L, 10-7 mol/L, 10-6 mol/L and 10-5 mol/L), and Wnt3a, LRP5 andβ-catenin mRNA expression were detect-ed using RT-PCR after 24 h, 48 h and 72 h. Results The cultured cells had the typical features of osteoblasts. Compared with that in control group, the result of RT-PCR showed that the expression of Wnt3a [(2.09±0.12) vs (1.82±0.09)], LRP5 [(1.42±0.07) vs (1.29±0.07)] andβ-catenin [(1.10±0.06) vs (0.95±0.06)] mRNA in 10-5 mol/L simvastatin group treat-ed after 24 h, and those of Wnt3a (F=37.78 and 96.52, P=0.000) and LRP5 (F=10.32, P=0.001 and F=22.02, P=0.000) increased significantly with the increase of concentration of simvastatin in each group treated after 48 h and 72 h (P<0.05). After 24 h, 48 h and 72 h intervention, the expression of Wnt3a and LRP5 mRNA increased significantly in each group as the intervention time got longer. In group 10-6 mol/L and 10-5 mol/L, compared with that after simvas-tatin intervention 48 h,β-catenin mRNA expression increased significantly after 72 h [(1.32±0.07) vs (1.11±0.05) and (1.47 ± 0.08) vs (1.19 ± 0.04), P<0.05]. Conclusion Simvastatin could promote Wnt3a, LRP5 andβ-catenin mRNA gene's expression of canonical Wnt signaling pathway in osteoblast of rats.
