本文選題：成纖維細(xì)胞　切入點(diǎn)：微波　出處：《第二軍醫(yī)大學(xué)》2008年碩士論文

【摘要】： 目的研究微波輻射對(duì)原代培養(yǎng)的小鼠皮膚成纖維細(xì)胞增殖活性的影響,進(jìn)而分析微波輻照后成纖維細(xì)胞的基因及蛋白的變化,探討微波對(duì)傷口修復(fù)階段瘢痕形成的影響。方法建立小鼠皮膚成纖維細(xì)胞原代培養(yǎng)的方法,經(jīng)不同強(qiáng)度微波直接輻射細(xì)胞不同時(shí)間后,用MTT法測(cè)定成纖維細(xì)胞的生長代謝,用流式細(xì)胞術(shù)監(jiān)測(cè)細(xì)胞周期分布。采用Real-time PCR擴(kuò)增的方法分析微波輻照后對(duì)細(xì)胞Ⅰ、Ⅲ型前膠原mRNA表達(dá)水平的影響。采用抗磷酸化形式的細(xì)胞外調(diào)節(jié)蛋白激酶抗體,通過免疫熒光染色方法和western blot法觀察成纖維細(xì)胞蛋白質(zhì)磷酸化情況。結(jié)果①成纖維細(xì)胞經(jīng)三種強(qiáng)度(0.5W/cm~2、1.0W/cm~2、5W/cm~2),不同時(shí)間(5min、15min、30min)微波輻射,其生長代謝曲線隨時(shí)間延長而下降。二種不同強(qiáng)度的微波(0.5W/cm~2、1.0W/cm~2)分別輻射5min、15min、30min,其成纖維細(xì)胞活性與假輻射組相比變化不大(P＞0.05)。強(qiáng)度5W/cm~2微波輻射5min以上即抑制成纖維細(xì)胞的增殖,細(xì)胞活性較假輻射組明顯降低(P＜0.05)。②強(qiáng)度5W/cm~2的微波輻射5min后,G0/G1期細(xì)胞所占比例明顯升高(P＜0.05)。③微波輻照對(duì)Ⅰ型前膠原基因mRNA表達(dá)的抑制作用隨時(shí)間的延長而增強(qiáng),在強(qiáng)度為1W/cm~2輻照30min、5W/cm~2輻照5min以上時(shí)能顯著抑制Ⅰ型前膠原基因mRNA的表達(dá)(P＜0.05);而對(duì)Ⅲ型前膠原mRNA表達(dá)水平的影響不大,但是Ⅰ型前膠原基因與Ⅲ型前膠原基因mRNA的比率明顯下降(P＜0.05);微波輻射后的成纖維細(xì)胞中MMP-9、MMP-13的mRNA表達(dá)水平均明顯增高。④體外短時(shí)間高強(qiáng)度微波輻射能夠使~pERK-1/2表達(dá)水平明顯升高。結(jié)論體外短時(shí)間高強(qiáng)度微波輻射能降低小鼠成纖維細(xì)胞的增殖活性,阻斷細(xì)胞周期,并使細(xì)胞停滯于G0/G1期,并且微波對(duì)成纖維細(xì)胞的增殖及其膠原產(chǎn)生的某些相關(guān)基因的表達(dá)具有直接的抑制作用。提示微波在創(chuàng)傷愈合的瘢痕形成階段起到減少瘢痕形成的作用。此外,微波輻射可激活皮膚成纖維細(xì)胞~pERK-1/2信號(hào)轉(zhuǎn)導(dǎo)通路,提示MAPK信號(hào)轉(zhuǎn)導(dǎo)通路參與了微波輻射對(duì)成纖維功能的調(diào)節(jié)過程。
[Abstract]:Objective to study the effect of microwave irradiation on the proliferation activity of primary cultured mouse skin fibroblasts, and to analyze the changes of gene and protein of fibroblasts after microwave irradiation, and to explore the effect of microwave on scar formation in wound repair stage.Methods the primary culture method of mouse skin fibroblasts was established. The growth and metabolism of fibroblasts were measured by MTT method and the cell cycle distribution was monitored by flow cytometry after microwave direct radiation of different intensity for different time.The effect of microwave irradiation on the expression of type 鈪，

本文編號(hào)：1712926