CAJ | 학술논문

干细胞%脂肪干细胞%多向诱导分化%条件培养液%口腔黏膜成纤维细胞%血管内皮生长因子%血小板源性生长因子%碱性成纤维细胞生长因子%蛋白芯片分析%国家自然科学基金%干细胞图片文章榦細胞%脂肪榦細胞%多嚮誘導分化%條件培養液%口腔黏膜成纖維細胞%血管內皮生長因子%血小闆源性生長因子%堿性成纖維細胞生長因子%蛋白芯片分析%國傢自然科學基金%榦細胞圖片文章
간세포%지방간세포%다향유도분화%조건배양액%구강점막성섬유세포%혈관내피생장인자%혈소판원성생장인자%감성성섬유세포생장인자%단백심편분석%국가자연과학기금%간세포도편문장

　　背景：近年来脂肪干细胞被证实对软组织创伤修复具有促进作用，关于其机制的研究中，更多的学者倾向于脂肪干细胞的旁分泌机制，即脂肪干细胞分泌的各种细胞因子对创伤部位细胞的修复功能具有促进作用。但脂肪干细胞分泌的因子种类，每种因子的含量，以及对软组织创伤修复，尤其是口腔黏膜的作用，鲜有报道。目的：观察脂肪干细胞条件培养液中血管内皮生长因子、血小板源性生长因子以及碱性成纤维细胞生长因子对口腔黏膜成纤维细胞增殖的影响。方法：采用蛋白芯片技术，检测脂肪干细胞条件培养液中血管内皮生长因子、血小板源性生长因子以及碱性成纤维细胞生长因子的含量。分别于1，2，3，4，5 d时，采用CCK-8法检测0，1，10，20，50，100μg/L血管内皮生长因子、血小板源性生长因子以及碱性成纤维细胞生长因子以及在脂肪干细胞条件培养液中分别加入上述各因子的中和抗体后，对口腔黏膜成纤维细胞增殖的影响。结果与结论：脂肪干细胞条件培养液中血管内皮生长因子、血小板源性生长因子以及碱性成纤维细胞生长因子信号值均大于300。脂肪干细胞条件培养液对成纤维细胞的增殖有明显的促进作用，其中，血小板源性生长因子和碱性成纤维细胞生长因子的促进作用非常明显，且随着因子浓度的变化而呈现峰值性改变，血小板源性生长因子的促进作用在50μg/L时达到峰值，碱性成纤维细胞生长因子的促进作用在1μg/L时达到峰值(P ≤0.05)，两者的中和抗体均能抑制脂肪干细胞条件培养液对口腔黏膜成纤维细胞的促进作用；而血管内皮细胞生长因子对成纤维细胞增殖无明显的促进作用。实验结果提示脂肪干细胞条件培养液可促进口腔黏膜成纤维细胞的增殖，其中所含有的血小板源性生长因子和碱性成纤维细胞生长因子的促进作用非常明显，且具有浓度依赖性，因此应注意选择适宜的细胞因子浓度，从而有效地促进成纤维细胞增殖。　　揹景：近年來脂肪榦細胞被證實對軟組織創傷脩複具有促進作用，關于其機製的研究中，更多的學者傾嚮于脂肪榦細胞的徬分泌機製，即脂肪榦細胞分泌的各種細胞因子對創傷部位細胞的脩複功能具有促進作用。但脂肪榦細胞分泌的因子種類，每種因子的含量，以及對軟組織創傷脩複，尤其是口腔黏膜的作用，鮮有報道。目的：觀察脂肪榦細胞條件培養液中血管內皮生長因子、血小闆源性生長因子以及堿性成纖維細胞生長因子對口腔黏膜成纖維細胞增殖的影響。方法：採用蛋白芯片技術，檢測脂肪榦細胞條件培養液中血管內皮生長因子、血小闆源性生長因子以及堿性成纖維細胞生長因子的含量。分彆于1，2，3，4，5 d時，採用CCK-8法檢測0，1，10，20，50，100μg/L血管內皮生長因子、血小闆源性生長因子以及堿性成纖維細胞生長因子以及在脂肪榦細胞條件培養液中分彆加入上述各因子的中和抗體後，對口腔黏膜成纖維細胞增殖的影響。結果與結論：脂肪榦細胞條件培養液中血管內皮生長因子、血小闆源性生長因子以及堿性成纖維細胞生長因子信號值均大于300。脂肪榦細胞條件培養液對成纖維細胞的增殖有明顯的促進作用，其中，血小闆源性生長因子和堿性成纖維細胞生長因子的促進作用非常明顯，且隨著因子濃度的變化而呈現峰值性改變，血小闆源性生長因子的促進作用在50μg/L時達到峰值，堿性成纖維細胞生長因子的促進作用在1μg/L時達到峰值(P ≤0.05)，兩者的中和抗體均能抑製脂肪榦細胞條件培養液對口腔黏膜成纖維細胞的促進作用；而血管內皮細胞生長因子對成纖維細胞增殖無明顯的促進作用。實驗結果提示脂肪榦細胞條件培養液可促進口腔黏膜成纖維細胞的增殖，其中所含有的血小闆源性生長因子和堿性成纖維細胞生長因子的促進作用非常明顯，且具有濃度依賴性，因此應註意選擇適宜的細胞因子濃度，從而有效地促進成纖維細胞增殖。
　　배경：근년래지방간세포피증실대연조직창상수복구유촉진작용，관우기궤제적연구중，경다적학자경향우지방간세포적방분비궤제，즉지방간세포분비적각충세포인자대창상부위세포적수복공능구유촉진작용。단지방간세포분비적인자충류，매충인자적함량，이급대연조직창상수복，우기시구강점막적작용，선유보도。목적：관찰지방간세포조건배양액중혈관내피생장인자、혈소판원성생장인자이급감성성섬유세포생장인자대구강점막성섬유세포증식적영향。방법：채용단백심편기술，검측지방간세포조건배양액중혈관내피생장인자、혈소판원성생장인자이급감성성섬유세포생장인자적함량。분별우1，2，3，4，5 d시，채용CCK-8법검측0，1，10，20，50，100μg/L혈관내피생장인자、혈소판원성생장인자이급감성성섬유세포생장인자이급재지방간세포조건배양액중분별가입상술각인자적중화항체후，대구강점막성섬유세포증식적영향。결과여결론：지방간세포조건배양액중혈관내피생장인자、혈소판원성생장인자이급감성성섬유세포생장인자신호치균대우300。지방간세포조건배양액대성섬유세포적증식유명현적촉진작용，기중，혈소판원성생장인자화감성성섬유세포생장인자적촉진작용비상명현，차수착인자농도적변화이정현봉치성개변，혈소판원성생장인자적촉진작용재50μg/L시체도봉치，감성성섬유세포생장인자적촉진작용재1μg/L시체도봉치(P ≤0.05)，량자적중화항체균능억제지방간세포조건배양액대구강점막성섬유세포적촉진작용；이혈관내피세포생장인자대성섬유세포증식무명현적촉진작용。실험결과제시지방간세포조건배양액가촉진구강점막성섬유세포적증식，기중소함유적혈소판원성생장인자화감성성섬유세포생장인자적촉진작용비상명현，차구유농도의뢰성，인차응주의선택괄의적세포인자농도，종이유효지촉진성섬유세포증식。
Adipose stem cel s have been confirmed to promote the repair of soft tissue after damage, and the action mechanism is possibly related to the paracrine of adipose stem cel s, that is adipose stem cel s secrete a variety of cytokines, which may promote the restoration of damaged cel s. However, little report addressed the types of adipose stem cel s secreted factors, contents of each factor, and role in soft tissue repair after damage, especial y oral mucosa. To observe the influence of vascular endothelial growth factor, platelet-derived growth factor, and basic fibroblast growth factor in adipose stem cel-conditioned medium on the proliferation of oral mucosa fibroblasts. Protein microarray analysis was used to analyze the contents of vascular endothelial growth factor, platelet-derived growth factor, and basic fibroblast growth factor in adipose stem cel-conditioned medium. CCK8 analysis was used to analyze the effects of adipose stem cel-conditioned medium with different concentrations of vascular endothelial growth factor, platelet-derived growth factor, and basic fibroblast growth factor (0, 1, 10, 20, 50 and 100μg/L) or the neutralizing antibody of the three kinds of cytokines on the proliferation of oral mucosa fibroblast on 1, 2, 3, 4 and 5 days. High contents of vascular endothelial growth factor, platelet-derived growth factor, and basic fibroblast growth factor were contained in adipose stem cel-conditioned medium (signal value>300). Adipose stem cel-conditioned medium could promote the proliferation of oral mucosa fibroblasts, wherein, the promotion effects of platelet-derived growth factor and basic fibroblast growth factor were very significant, and the peak changes could be observed with variation of cytokines concentrations. The optimal concentrations of platelet-derived growth factor and basic fibroblast growth factor were 50μg/L and 1μg/L, respectively (P ≤ 0.05). The neutralizing antibody of platelet-derived growth factor and basic fibroblast growth factor inhibited the promotion effect of adipose stem cel-conditioned medium. On the other hand, vascular endothelial growth factor had no significant effect on the proliferation of oral mucosa fibroblasts. Adipose stem cel-conditioned medium can promote the proliferation of oral mucosa fibroblasts, platelet-derived growth factor and basic fibroblast growth factor in adipose stem cel-conditioned medium have obvious promotion effects, which are dependent on the cytokine concentrations. Therefore, we should pay attention to choose the optimal concentrations of cytokines, thereby effectively promoting the proliferation of fibroblasts.