世界中医药
世界中醫藥
세계중의약
World Chinese Medicine
2015年
10期
1578-1581
,共4页
接骨丹%含药血清%成骨细胞%骨折愈合
接骨丹%含藥血清%成骨細胞%骨摺愈閤
접골단%함약혈청%성골세포%골절유합
Bone-Joining Elixir%Medicated Serum%Osteoblast%Fracture Healing
目的：观察接骨丹对新西兰乳兔成骨细胞增殖的影响，探讨接骨丹促进骨折愈合的机制。方法：普通级新西兰大白乳兔，耳缘静脉空气栓塞处死，无菌条件下取颅盖骨，Ⅰ型胶原酶消化法获取成骨细胞并进行体外培养、鉴定及传代。将鉴定后的第二代成骨细胞随机分为正常对照组及实验组，正常对照组应用正常兔血清培养，实验组应用接骨丹含药血清培养，MTT 法观测2组细胞的生长曲线；不同血清干预48 h 后，倒置显微镜观测接骨丹对成骨细胞碱性磷酸酶及Ⅰ型胶原表达的影响，Elisa 法观测接骨丹对成骨细胞培养上清液中骨钙素表达的影响。结果：Ⅰ型胶原酶消化法成功获取并建立成骨细胞体外培养体系，MTT 观测显示实验组能够显著促进成骨细胞的增值，倒置显微镜观测实验组中碱性磷酸酶及Ⅰ型胶原的表达要优于正常对照组，并能够促进成骨细胞分泌骨钙素。结论：接骨丹能够促进骨折的愈合，其机制可能与促进成骨细胞中碱性磷酸酶、Ⅰ型胶原及骨钙素的表达进而促进成骨细胞增殖和分化有关。
目的：觀察接骨丹對新西蘭乳兔成骨細胞增殖的影響，探討接骨丹促進骨摺愈閤的機製。方法：普通級新西蘭大白乳兔，耳緣靜脈空氣栓塞處死，無菌條件下取顱蓋骨，Ⅰ型膠原酶消化法穫取成骨細胞併進行體外培養、鑒定及傳代。將鑒定後的第二代成骨細胞隨機分為正常對照組及實驗組，正常對照組應用正常兔血清培養，實驗組應用接骨丹含藥血清培養，MTT 法觀測2組細胞的生長麯線；不同血清榦預48 h 後，倒置顯微鏡觀測接骨丹對成骨細胞堿性燐痠酶及Ⅰ型膠原錶達的影響，Elisa 法觀測接骨丹對成骨細胞培養上清液中骨鈣素錶達的影響。結果：Ⅰ型膠原酶消化法成功穫取併建立成骨細胞體外培養體繫，MTT 觀測顯示實驗組能夠顯著促進成骨細胞的增值，倒置顯微鏡觀測實驗組中堿性燐痠酶及Ⅰ型膠原的錶達要優于正常對照組，併能夠促進成骨細胞分泌骨鈣素。結論：接骨丹能夠促進骨摺的愈閤，其機製可能與促進成骨細胞中堿性燐痠酶、Ⅰ型膠原及骨鈣素的錶達進而促進成骨細胞增殖和分化有關。
목적：관찰접골단대신서란유토성골세포증식적영향，탐토접골단촉진골절유합적궤제。방법：보통급신서란대백유토，이연정맥공기전새처사，무균조건하취로개골，Ⅰ형효원매소화법획취성골세포병진행체외배양、감정급전대。장감정후적제이대성골세포수궤분위정상대조조급실험조，정상대조조응용정상토혈청배양，실험조응용접골단함약혈청배양，MTT 법관측2조세포적생장곡선；불동혈청간예48 h 후，도치현미경관측접골단대성골세포감성린산매급Ⅰ형효원표체적영향，Elisa 법관측접골단대성골세포배양상청액중골개소표체적영향。결과：Ⅰ형효원매소화법성공획취병건립성골세포체외배양체계，MTT 관측현시실험조능구현저촉진성골세포적증치，도치현미경관측실험조중감성린산매급Ⅰ형효원적표체요우우정상대조조，병능구촉진성골세포분비골개소。결론：접골단능구촉진골절적유합，기궤제가능여촉진성골세포중감성린산매、Ⅰ형효원급골개소적표체진이촉진성골세포증식화분화유관。
Objective:To observe the effect of Bone-Joining Elixir on New Zealand newborn rabbit osteoblast proliferation,and ex-plore the promoting fracture healing mechanism of Bone-Joining Elixir.Methods:Conventional grade New Zealand big and white newborn rabbits were put to death by ear venous air embolism,the craniums were taken down in aseptic conditions,and the osteo-blasts were gained by collagenase type Ⅰ digestion method for in vitro culture,identification and passages.After identification, the second generation of osteoblasts was randomly divided into normal control group and experimental group.And the normal con-trol group was cultured in normal rabbit serum,the experimental group was cultured in Bone-Joining Elixir medicated serum,MTT method was taken to observe the cell growth curves of the two groups.After the intervention for 48 h,the two serums were observed to achieve the influence of Bone-Joining Elixir on the expression of osteoblast alkaline phosphatase and collagen typeⅠ under the inverted microscope,and to achieve the influence of Bone-Joining Elixir on the expression of osteocalcin in the supernate in the os-teoblast culture by Elisa method.Results:The osteoblast culture system in vitro has been successfully obtained and established by the collagenase type Ⅰ digestion method,MTT observation indicated the experimental group show significant promotion on the pro-liferation of osteoblasts,and the inverted microscope indicated the expression of alkaline phosphatase and collagen type Ⅰ in the experimental group was superior to the normal control group,and the osteoblast was promoted to secrete osteocalcin.Conclusion:The Bone-Joining Elixir can promote the healing of fracture,the mechanism may be related to the promotion of the expression of al-kaline phosphatase,collagen type Ⅰ and osteocalcin in osteoblasts to help osteoblasts proliferate and differentiate.
