本文選題：堿性成纖維細胞生長因子 + 角膜上皮細胞��； 參考：《華中農(nóng)業(yè)大學》2007年碩士論文

【摘要】： 堿性成纖維細胞生長因子(bFGF)是一種廣譜促有絲分裂原，對幾乎所有來源于中胚層和神經(jīng)外胚層的細胞具有廣泛的生物學活性，它具有良好地神經(jīng)保護、心肌保護、缺血保護以及促進創(chuàng)面愈合作用，使其臨床治療日益受到關(guān)注，同時bFGF在眼科領(lǐng)域的研究也日益深入。 制作角膜上皮細胞，用DMEM／F-12培養(yǎng)液培養(yǎng)，將培養(yǎng)到第三代的角膜上皮細胞用四甲基偶氮唑鹽(MTT)比色法觀察堿性成纖維細胞生長因子在不同濃度(0ng／mL，1ng／mL，10ng／mL，，100ng／mL)和不同時間點(24h，48h，96h)對犬角膜上皮細胞培養(yǎng)增殖性能來研究堿性成纖維細胞生長因子對犬角膜上皮細胞的生長和增殖的影響，檢測所的的結(jié)果表示為光密度值(OD)值，OD值的大小與細胞密度呈正比。結(jié)果表明，從1ng／mL開始，隨濃度遞增，OD值逐漸上升，10ng／mL時達到峰值，此后隨濃度遞增OD值下降。bFGF組從1ng／mL開始各濃度組OD值均大于對照組(P＜0.05)，說明bFGF對犬角膜上皮細胞有明顯的促分裂作用。10ng／mL組在24h、48h、96h均高于其他劑量組。說明堿性成纖維細胞生長因子有明顯促進犬角膜上皮細胞增殖的作用，在一定范圍內(nèi)呈劑量依賴性，10ng／mL為促進犬角膜上皮生長的最佳劑量。 此外，本研究建立了犬角膜環(huán)鉆損傷模型，分析堿性成纖維細胞生長因子對犬角膜損傷的治療效果。將24只雜交犬隨機分成四組：生理鹽水對照組、0.1μg／mL bFGF組、0.5μg／mLbFGF組、1μg／mLbFGF組。每組使用不同濃度的bFGF點眼，每日三次，每次兩滴。并在術(shù)后第1d、第3d、第5d、第7d同一時間用熒光染色素鈉染液點眼，照相觀察角膜上皮愈合效果。并于術(shù)后第3d、第7d、第14d取角膜各5只制作病理切片觀察角膜上皮愈合情況及膠原纖維排列情況、成纖維細胞生長情況。熒光染色結(jié)果表明，環(huán)鉆損傷后第1 d，0.5μg／mL劑量組愈合率為27.47％，1μg／mL劑量組愈合率為31.23％，與對照組相比差異極顯著(P＜0.01)。環(huán)鉆損傷后第3d、第5d不同濃度的bFGF各組愈合率均極顯著高于對照組(P＜0.01)，第7d時除對照組有兩例愈合不良外各組均基本愈合。顯示堿性成纖維細胞生長因子能促進犬角膜損傷后角膜上皮愈合，早期應用即能發(fā)揮作用。病理切片觀察發(fā)現(xiàn)添加堿性成纖維細胞生長因子的犬角膜上皮修復快于對照組，表現(xiàn)為細胞層數(shù)比對照組多，細胞排列整齊，焱性細胞浸潤少，另外堿性成纖維細胞生長因子還能促進犬角膜損傷后角膜基質(zhì)有序修復。未發(fā)現(xiàn)堿性成纖維細胞生長因子對犬角膜損傷后角膜新生血管生長有明顯促進或抑制作用。
[Abstract]:Basic fibroblast growth factor (bFGF) is a broad-spectrum mitogen with extensive biological activity to almost all cells from mesoderm and neuroectoderm. The protection of ischemia and the promotion of wound healing have attracted more and more attention in clinical treatment. Meanwhile, the research of bFGF in ophthalmology has been deepened. Corneal epithelial cells were prepared and cultured in DMEM/F-12 medium. The third passage corneal epithelial cells were cultured by MTT colorimetry to study the effect of basic fibroblast growth factor (BFGF) on the proliferation of canine corneal epithelial cells at different concentrations (0 ng / mL ~ (-1) 10 ng / mL ~ (-1) ~ 100 ng / m ~ (-1) and at 24 h / m ~ (-1) for 48 h / 96 h) to study the effect of alkaloids on the proliferation of canine corneal epithelial cells in vitro. Effects of fibroblast growth factor on the growth and proliferation of canine corneal epithelial cells The results show that the OD value is proportional to the cell density. The results showed that from the beginning of 1ng/mL, the OD value gradually increased to a peak value of 10 ng / mL with increasing concentration. From then on, the OD value of each concentration group was higher than that of the control group (P < 0.05) from the beginning of 1ng/mL, which indicated that bFGF had obvious mitogenic effect on canine corneal epithelium cells. 10 ng / mL group was significantly higher than other dose groups at 24 h and 48 h to 96 h. The results showed that basic fibroblast growth factor (bFGF) could obviously promote the proliferation of canine corneal epithelial cells. In a dose-dependent manner, 10 ng / mL was the best dose to promote the growth of canine corneal epithelium. In addition, a canine corneal annular injury model was established, and the therapeutic effect of basic fibroblast growth factor (bFGF) on canine corneal injury was analyzed. 24 crossbred dogs were randomly divided into four groups: saline control group, 0. 1 渭 g/mL bFGF group, 0. 5 渭 g/mLbFGF group and 1 渭 g/mLbFGF group. Each group was given two drops of different concentrations of bFGF three times a day. The corneal epithelium healing was observed with fluorescein sodium at the same time on the 1st, 3rd, 5th and 7th day after operation. At the 3rd, 7th and 14th day, the corneal epithelium healing, collagen fiber arrangement and fibroblast growth were observed. The results of fluorescence staining showed that the healing rate of the 0.5 渭 g/mL group was 27.47 and the healing rate of the 1 渭 g/mL group was 31.233.The difference was significant compared with the control group (P < 0.01). The healing rate of different concentrations of bFGF was significantly higher than that of the control group on the 3rd and 5th day after drilling injury (P < 0.01). On the 7th day, there were two cases of poor healing in the control group. The results showed that basic fibroblast growth factor (bFGF) could promote corneal epithelial healing after corneal injury in dogs. The results of pathological section showed that the corneal epithelium repaired by adding basic fibroblast growth factor was faster than that of the control group, which showed that the number of cell layers was more than that of the control group, the cells were arranged neatly, and the infiltration of Yan cells was less than that of the control group. In addition, basic fibroblast growth factor can also promote corneal stroma repair after corneal injury in dogs. It was not found that basic fibroblast growth factor could significantly promote or inhibit corneal neovascularization after corneal injury in dogs.
【學位授予單位】：華中農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2007
【分類號】：R341

【相似文獻】

相關(guān)期刊論文 前10條

1 鄧蔡;陳建鋼;龐光明;;bFGF對人牙髓、牙周膜成纖維細胞生物學效應的研究[J];臨床口腔醫(yī)學雜志;2011年08期

2 衛(wèi)秀洋;王萬明;陳慶泉;;堿性成纖維細胞生長因子在組織修復的研究進展[J];中國矯形外科雜志;2011年13期

3 黃生高;王月輝;周sビ

本文編號：1863059