中华内分泌代谢杂志
中華內分泌代謝雜誌
중화내분비대사잡지
CHINESE JOURNAL OF ENDOCRINOLOGY AND METABOLISM
2010年
6期
489-492
,共4页
宋利格%张秀珍%赵家胜%雷涛%贺铭%张春阳%周筠
宋利格%張秀珍%趙傢勝%雷濤%賀銘%張春暘%週筠
송리격%장수진%조가성%뢰도%하명%장춘양%주균
淫羊藿甙%黄酮类%成骨细胞%核心结合因子α1%丝裂原活化蛋白激酶
淫羊藿甙%黃酮類%成骨細胞%覈心結閤因子α1%絲裂原活化蛋白激酶
음양곽대%황동류%성골세포%핵심결합인자α1%사렬원활화단백격매
Icariin%Flavones%Osteoblasts%Core binding factor-α1%Mitogen-activated protein kinase
目的 观察淫羊藿甙对成骨细胞中核心结合因子α1(cbfa1)蛋白和活性表达的调节,以及丝裂原活化蛋白激酶(MAPK)信号通路是否参与此过程.方法 用酶消化法分离24 h内新生SD大鼠颅盖骨成骨细胞,进行原代培养,经鉴定后用于实验.设立对照组、淫羊藿甙组(10 ng/ml)以及雌二醇组(10-8mol/L),分别用药物干预24 h,抽提核蛋白.利用转录因子活性ELISA法检测成骨细胞cbfa1与DNA结合的活性,Western印迹法检测成骨细胞中cbfa1蛋白的表达.将MAPK信号转导通路抑制剂U0126和SB203580分别与淫羊藿甙或雌二醇共同加入培养液中,培养24 h,同上法检测Cbfa1活性和Cbh1蛋白表达量的变化.结果 淫羊藿甙和雌二醇均可以促进成骨细胞中Cbfa1活性和Cbh1蛋白表达量的提高(P<0.05).加入细胞外信号调节激酶(ERK)途径的抑制剂UO126后,可以下调淫羊藿甙和雌二醇对成骨细胞中Cbfa1活性和Cbh1蛋白表达量的上调作用(P<0.05).加入p38MAPK途径的抑制剂SB203580后,也可以下调淫羊藿甙和雌二醇对成骨细胞中Cbfa1活性和Cbh1蛋白表达量的上调作用(P<0.05).结论 淫羊藿甙和雌激素均可上调体外培养大鼠成骨细胞转录因子Cbh1的蛋白表达和结合活性.MAPK信号转导通路抑制剂可以部分阻断淫羊藿甙和雌激素对成骨细胞转录因子Cbh1蛋白表达和活性的上调作用,说明MAPK可能是淫羊藿甙发挥抗骨质疏松作用的信号转导通路之一.
目的 觀察淫羊藿甙對成骨細胞中覈心結閤因子α1(cbfa1)蛋白和活性錶達的調節,以及絲裂原活化蛋白激酶(MAPK)信號通路是否參與此過程.方法 用酶消化法分離24 h內新生SD大鼠顱蓋骨成骨細胞,進行原代培養,經鑒定後用于實驗.設立對照組、淫羊藿甙組(10 ng/ml)以及雌二醇組(10-8mol/L),分彆用藥物榦預24 h,抽提覈蛋白.利用轉錄因子活性ELISA法檢測成骨細胞cbfa1與DNA結閤的活性,Western印跡法檢測成骨細胞中cbfa1蛋白的錶達.將MAPK信號轉導通路抑製劑U0126和SB203580分彆與淫羊藿甙或雌二醇共同加入培養液中,培養24 h,同上法檢測Cbfa1活性和Cbh1蛋白錶達量的變化.結果 淫羊藿甙和雌二醇均可以促進成骨細胞中Cbfa1活性和Cbh1蛋白錶達量的提高(P<0.05).加入細胞外信號調節激酶(ERK)途徑的抑製劑UO126後,可以下調淫羊藿甙和雌二醇對成骨細胞中Cbfa1活性和Cbh1蛋白錶達量的上調作用(P<0.05).加入p38MAPK途徑的抑製劑SB203580後,也可以下調淫羊藿甙和雌二醇對成骨細胞中Cbfa1活性和Cbh1蛋白錶達量的上調作用(P<0.05).結論 淫羊藿甙和雌激素均可上調體外培養大鼠成骨細胞轉錄因子Cbh1的蛋白錶達和結閤活性.MAPK信號轉導通路抑製劑可以部分阻斷淫羊藿甙和雌激素對成骨細胞轉錄因子Cbh1蛋白錶達和活性的上調作用,說明MAPK可能是淫羊藿甙髮揮抗骨質疏鬆作用的信號轉導通路之一.
목적 관찰음양곽대대성골세포중핵심결합인자α1(cbfa1)단백화활성표체적조절,이급사렬원활화단백격매(MAPK)신호통로시부삼여차과정.방법 용매소화법분리24 h내신생SD대서로개골성골세포,진행원대배양,경감정후용우실험.설립대조조、음양곽대조(10 ng/ml)이급자이순조(10-8mol/L),분별용약물간예24 h,추제핵단백.이용전록인자활성ELISA법검측성골세포cbfa1여DNA결합적활성,Western인적법검측성골세포중cbfa1단백적표체.장MAPK신호전도통로억제제U0126화SB203580분별여음양곽대혹자이순공동가입배양액중,배양24 h,동상법검측Cbfa1활성화Cbh1단백표체량적변화.결과 음양곽대화자이순균가이촉진성골세포중Cbfa1활성화Cbh1단백표체량적제고(P<0.05).가입세포외신호조절격매(ERK)도경적억제제UO126후,가이하조음양곽대화자이순대성골세포중Cbfa1활성화Cbh1단백표체량적상조작용(P<0.05).가입p38MAPK도경적억제제SB203580후,야가이하조음양곽대화자이순대성골세포중Cbfa1활성화Cbh1단백표체량적상조작용(P<0.05).결론 음양곽대화자격소균가상조체외배양대서성골세포전록인자Cbh1적단백표체화결합활성.MAPK신호전도통로억제제가이부분조단음양곽대화자격소대성골세포전록인자Cbh1단백표체화활성적상조작용,설명MAPK가능시음양곽대발휘항골질소송작용적신호전도통로지일.
Objective To investigate the effects of icarrin on the activity and protein expression of core binding factor otl(Cbfa1) in rat osteoblasts cultured in vitro,and to explore whether mitogen-activated protein kinase (MAPK) pathway is involved in this process.Methods Calvarial osteoblasts were obtained from newborn (<24 h) SD rats by trypsin-coUagenase digestion method.The second generation osteoblasts were cultured in the medium containing icariin (10 ng/ml) or estradiol (10-8 mol/L) with or without extracellular-signal regulated kinase (ERK) inhibitor (UO126) or p38MAPK inhibitor (SB203580).Nuclear protein was extracted from osteoblasts.And then the activity of Cbfa1 was detected by ELISA.The amounts of Cbfa1 protein were detected by Western blot.Results Calvarial osteoblasts were obtained successfully and were used in this study after indentified by alkaline phosphatase and mineralized nodus staining.Cbfa1 expression and the activity in osteoblasts were up-regulated by both icariin and estradiol (P<0.05).The effects were partly inhibited by addition of U0126or SB203580 (P<0.05).Conclusions Either icarrin or estradiol can stimulate the proliferation and maturation of cultured osteoblasts in vitro via up-regulating the activity and expression of Cbfal.The MAPK signal pathway inhibitor seems to partly decrease Cbfa1 activity.It suggests that MAPK pathway may be involved in the transduction of icariin's impact on proliferation and mineralization of osteoblasts.
