【摘要】： 目的：研究不同濃度的CTGF對體外培養(yǎng)肌腱細胞增殖的影響，探討CTGF參與肌腱損傷修復的可能機制。 方法：本實驗以成年兔后肢趾屈肌腱為材料，，用酶消化后，將分離的細胞用含10％胎牛血清的培養(yǎng)基培養(yǎng)，貼壁達80％后傳代。觀察細胞的形態(tài)及生長規(guī)律。取第二代細胞，通過免疫組化的方法進行肌腱細胞鑒定。取第三代肌腱細胞，按細胞數(shù)約為4×10~3／孔接種在96孔培養(yǎng)板的48孔，每6孔為一組，共分為8組，根據(jù)每組CTGF濃度不同，分為5ng／ml CTGF組、10ng／ml CTGF組、20ng／ml CTGF組、50ng／ml CTGF組、100ng／ml CTGF組、200ng／ml CTGF組、500ng／ml CTGF組及對照組(不加CTGF)。顯微鏡下觀察各組細胞均進入對數(shù)生長期時，用噻唑藍(methyl thiazolyl tetrazolium，MTT)法染色，在酶聯(lián)免疫檢測儀上測出各孔吸光度(optical density，OD)值，并計算各組OD均值進行比較。 結(jié)果：MTT實驗通過比較各組OD值發(fā)現(xiàn)：5ng／mlCTGF組肌腱細胞增殖與對照組無顯著性差異；5ng／ml、10ng／ml、20ng／ml、50ng／mlCTGF組肌腱細胞增殖有隨CTGF濃度增加而上升的趨勢，且各組之間的差異均有顯著性(P＜0.05)；50ng／ml、100ng／ml、200ng／ml CTGF、500ng／ml CTGF組肌腱細胞增殖有隨濃度增加而稍呈下降的趨勢，但各組之間的差異均無顯著性(P＞0.05)。 結(jié)論：10ng／mlCTGF、20ng／mlCTGF、50ng／mlCTGF、100ng／mlCTGF、200ng／mlCTGF、500ng／mlCTGF有促進肌腱細胞增殖作用；5ng／mlCTGF對肌腱細胞增殖無明顯作用。CTGF濃度在0ng／ml～50ng／ml之間時，肌腱細胞的增殖隨濃度增加呈上升趨勢；在50ng／ml～500ng／ml之間時，肌腱細胞的增殖隨濃度增加稍呈下降趨勢。50ng／ml為促肌腱細胞增殖的最佳CTGF濃度。
[Abstract]:Aim: to investigate the effects of different concentrations of CTGF on the proliferation of cultured tendon cells in vitro and to explore the possible mechanism of CTGF involved in tendon repair. Methods: after digested with enzyme, the isolated cells were cultured on the culture medium containing 10% fetal bovine serum. The morphology and growth of cells were observed. The second generation cells were identified by immunohistochemical method. The third passage of tendon cells were divided into 8 groups according to the number of cells (4 脳 10 ~ 3 / well) inoculated in the 96-well culture plate. According to the concentration of CTGF in each group, they were divided into 5ng/ml CTGF group, 10ng/ml CTGF group and 20ng/ml CTGF group. 50ng/ml CTGF group, 100ng/ml CTGF group, 200ng/ml CTGF group, 500ng/ml CTGF group and control group (without CTGF).) When the cells in each group entered logarithmic growth period, the absorbance (optical density,OD) of each hole was measured by using the enzyme linked immunosorbent assay (Elisa) with thiazolyl blue (methyl thiazolyl tetrazolium,MTT method, and the mean value of OD in each group was calculated and compared. Results: MTT test showed that there was no significant difference in the proliferation of tendon cells between the 5ng/mlCTGF group and the control group by comparing the OD values of each group. The proliferation of tendon cells in the 10ng / ml 10 ng / ml CTGF group increased with the increase of CTGF concentration, and there were significant differences among the groups (P < 0. 05). The proliferation of tendon cells in the 50ng / ml CTGF,500ng/ml CTGF group decreased slightly with the increase of the concentration, but there was no significant difference between the two groups (P > 0. 05). Conclusion: 10ng / ml CTGF1 20ng / ml CTGFF 50ng / ml CTGFG / 100ng / ml CTGFF 200ng / ml CTGF500ng / ml CTGF can promote the proliferation of tendon cells. 5ng/mlCTGF had no obvious effect on the proliferation of tendon cells. When the concentration of CTGF was between 0ng/ml~50ng/ml, the proliferation of tendon cells increased with the increase of 0ng/ml~50ng/ml concentration. Between 50ng/ml~500ng/ml and 50ng/ml~500ng/ml, the proliferation of tendon cells decreased slightly with the increase of CTGF concentration. 50ng/ml was the best CTGF concentration to promote the proliferation of tendon cells.
【學位授予單位】：中南大學
【學位級別】：碩士
【學位授予年份】：2007
【分類號】：R329

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