中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
10期
1496-1501
,共6页
孙月娇%宋囡%何文智%何丽娟%任艳玲
孫月嬌%宋囡%何文智%何麗娟%任豔玲
손월교%송닙%하문지%하려연%임염령
干细胞%骨髓干细胞%左归丸%右归丸%肾藏精%含药血清%骨髓间充质干细胞%成骨诱导%Ⅰ型胶原蛋白%信号转导%转化生长因子β1%信号转导蛋白%Smad2/3%国家自然科学基金
榦細胞%骨髓榦細胞%左歸汍%右歸汍%腎藏精%含藥血清%骨髓間充質榦細胞%成骨誘導%Ⅰ型膠原蛋白%信號轉導%轉化生長因子β1%信號轉導蛋白%Smad2/3%國傢自然科學基金
간세포%골수간세포%좌귀환%우귀환%신장정%함약혈청%골수간충질간세포%성골유도%Ⅰ형효원단백%신호전도%전화생장인자β1%신호전도단백%Smad2/3%국가자연과학기금
stem cells%mesenchymal stem cells%bone marrow%osteoblasts%adaptor proteins,signal transducing%transforming growth factors
背景：滋补肾阴左归丸、温补肾阳右归丸均可协同诱导剂诱导骨髓间充质干细胞向成骨细胞分化，但其机制尚有待探讨。<br> 目的：观察左、右归丸含药血清对体外培养的大鼠骨髓间充质干细胞向成骨细胞分化过程中转化生长因子β1及其信号转导蛋白Smad2/3表达的影响。<br> 方法：运用全骨髓贴壁法分离和培养大鼠骨髓间充质干细胞；分别以空白血清对照组，左归丸、右归丸、阳性对照药补佳乐制备的大鼠含药血清加诱导剂(地塞米松、维生素C、β-甘油磷酸钠)，以及诱导剂组为对照共5组对骨髓间充质干细胞进行干预。采用Western blot法检测Ⅰ型胶原蛋白表达，用免疫组化法检测转化生长因子和Smad2/3在细胞内的表达。<br> 结果与结论：左归丸、右归丸含药血清诱导骨髓间充质干细胞成骨后Ⅰ型胶原蛋白、转化生长因子β1、Smad2/3蛋白表达高于空白组、诱导剂组，差异有显著性意义(P <0.05)，且左归丸优于右归丸(P <0.05)。提示左、右归丸含药血清能通过转化生长因子β1调节Smad2/3信号通路促进骨髓间充质干细胞成骨分化，且左归丸组效果更好，表明中医“滋肾阴”法更能有效的促进骨髓间充质干细胞的成骨分化。
揹景：滋補腎陰左歸汍、溫補腎暘右歸汍均可協同誘導劑誘導骨髓間充質榦細胞嚮成骨細胞分化，但其機製尚有待探討。<br> 目的：觀察左、右歸汍含藥血清對體外培養的大鼠骨髓間充質榦細胞嚮成骨細胞分化過程中轉化生長因子β1及其信號轉導蛋白Smad2/3錶達的影響。<br> 方法：運用全骨髓貼壁法分離和培養大鼠骨髓間充質榦細胞；分彆以空白血清對照組，左歸汍、右歸汍、暘性對照藥補佳樂製備的大鼠含藥血清加誘導劑(地塞米鬆、維生素C、β-甘油燐痠鈉)，以及誘導劑組為對照共5組對骨髓間充質榦細胞進行榦預。採用Western blot法檢測Ⅰ型膠原蛋白錶達，用免疫組化法檢測轉化生長因子和Smad2/3在細胞內的錶達。<br> 結果與結論：左歸汍、右歸汍含藥血清誘導骨髓間充質榦細胞成骨後Ⅰ型膠原蛋白、轉化生長因子β1、Smad2/3蛋白錶達高于空白組、誘導劑組，差異有顯著性意義(P <0.05)，且左歸汍優于右歸汍(P <0.05)。提示左、右歸汍含藥血清能通過轉化生長因子β1調節Smad2/3信號通路促進骨髓間充質榦細胞成骨分化，且左歸汍組效果更好，錶明中醫“滋腎陰”法更能有效的促進骨髓間充質榦細胞的成骨分化。
배경：자보신음좌귀환、온보신양우귀환균가협동유도제유도골수간충질간세포향성골세포분화，단기궤제상유대탐토。<br> 목적：관찰좌、우귀환함약혈청대체외배양적대서골수간충질간세포향성골세포분화과정중전화생장인자β1급기신호전도단백Smad2/3표체적영향。<br> 방법：운용전골수첩벽법분리화배양대서골수간충질간세포；분별이공백혈청대조조，좌귀환、우귀환、양성대조약보가악제비적대서함약혈청가유도제(지새미송、유생소C、β-감유린산납)，이급유도제조위대조공5조대골수간충질간세포진행간예。채용Western blot법검측Ⅰ형효원단백표체，용면역조화법검측전화생장인자화Smad2/3재세포내적표체。<br> 결과여결론：좌귀환、우귀환함약혈청유도골수간충질간세포성골후Ⅰ형효원단백、전화생장인자β1、Smad2/3단백표체고우공백조、유도제조，차이유현저성의의(P <0.05)，차좌귀환우우우귀환(P <0.05)。제시좌、우귀환함약혈청능통과전화생장인자β1조절Smad2/3신호통로촉진골수간충질간세포성골분화，차좌귀환조효과경호，표명중의“자신음”법경능유효적촉진골수간충질간세포적성골분화。
BACKGROUND:Bone marrow mesenchymal stem cells can differentiate into osteoblasts under inducing condition that Zouguiwan and Youguiwan coordinate inducers, but the mechanism remains to be discussed. OBJECTIVE:To observe the effects of serum containing Zuoguiwan and Youguiwan on transforming growth factorβ1 and its signal transduction protein Smad2/3 message expression during the osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS:A whole bone marrow adherence method was adopted to isolate and cultivate bone marrow mesenchymal stem cells from rats. The cellcultivation was processed in five groups:bone marrow mesenchymal stem cells were respectively cultured with blank serum, serum containing Zouguiwan, serum containing Youguiwan, positive serum containing progynova+inducer (dexamethasone, vitamin C, andβ-glycerophosphate), and inducer. Western blot was applied to detect the expression of type I col agen. The immunohistochemical assay was utilized to test transforming growth factorβ1 and Smad2/3 expression in the osteoblasts. RESULTS AND CONCLUSION:It was apparently more significant for serum containing Zuoguiwan and Youguiwan on type I col agen, transforming growth factorβ1 and Smad2/3 expression, compared with blank serum group and inducer group (P<0.05);moreover, serum containing Zuoguiwan was better than serum containing Youguiwan (P<0.05). Both of serum containing Zuoguiwan and Youguiwan are able to promote osteogenic differentiation of bone marrow mesenchymal stem cells. Moreover, Zuoguiwan is much more effective indicating that this method of traditional Chinese medicine about nourishing kidneys can be better to promote osteogenic induction of bone marrow mesenchymal stem cells.
