發(fā)布時間：2018-06-15 10:28

  本文選題：G蛋白偶聯(lián)雌激素受體 + 增生性瘢痕��； 參考：《遵義醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:1.通過局部應(yīng)用雌素來觀察并驗證其對于兔耳增生性瘢痕的影響作用。2.研究G蛋白偶聯(lián)雌激素受體(GPER)介導(dǎo)的雌激素快速非基因組效應(yīng)對于兔耳增生性瘢痕的影響及作用機制。方法:選取3-6月齡新西蘭大耳兔,36只,體重2.5-3kg,雌性,要求兔耳皮膚完整無破損;適應(yīng)性飼養(yǎng)7天后,在兔的單側(cè)耳腹面共做6個直徑為10mm的圓形創(chuàng)面,間距大于15mm,深達軟骨層并刮除軟骨膜,建立兔耳增生性瘢痕模型。按照隨機數(shù)字表法將36只新西蘭大耳兔分為6個組,每組6只:(control組)為空白對照組;(DMSO組)為溶劑組;(E2組)為雌激素組;(G1組)為GPER激動劑組;(E2+G15組)為雌激素+GPER拮抗劑組;(G1+G15組)為GPER激動劑+拮抗劑組。于造模后第15天,4個給藥組分別用微量注射器在兔耳根部皮下注射E2 0.025mg/(kg·d),G1 0.06 mg/(kg·d),G15 0.06 mg/(kg·d),溶劑組注射經(jīng)PBS稀釋為10%的DMSO溶液,對照組不予任何處理。持續(xù)注射14天后,觀察各組瘢痕增生情況并取標(biāo)本,對比各組瘢痕增生指數(shù)(SEI)、成纖維細(xì)胞(FB)、膠原纖維以及通過PCR檢測TGF-β1、collagen III、collagen I的表達量。結(jié)果:1.形態(tài)學(xué)觀察:E2組和G1組相對于空白對照組,瘢痕突出周圍正常皮膚更為明顯,顏色較深多為暗紅色,中央部最為厚實,質(zhì)地較硬;E2+G15組和G1+G15組對比E2組和G1組,瘢痕中央厚度明顯降低,顏色也較淺為粉紅色,部分與正常皮膚接近,質(zhì)地明顯變軟;DMSO組相對于空白對照組瘢痕增生情況差別不大。2.HE染色和MASSON染色觀察:E2組和G1組相對于空白對照組,瘢痕組織中存在大量的成纖維細(xì)胞,血管生長豐富,細(xì)胞外基質(zhì)有大量沉積,膠原纖維多而粗大,排列紊亂。E2+G15組和G1+G15組對比E2組和G1組,成纖維細(xì)胞明顯減少,血管分布較少,細(xì)胞外基質(zhì)沉積減少,膠原纖維也相對減少,DMSO組相對于空白對照組膠原纖維、成纖維細(xì)胞和血管分布未見特殊異常。3.瘢痕增生指數(shù):E2組和G1組明顯高于空白對照組P0.05),E2+G15組相對于E2組明顯降低(P0.05),G1+G15組相對于G1組也明顯降低(P0.05),而空白對照組和DMSO組無明顯差異(P0.05)。4.RT-PCR檢測:與空白對照組相比,E2組和G1組的collagenⅠ、collagenⅢ、TGF-β1的m RNA表達量明顯升高(P0.05);而E2+G15組和G1+G15組分別對比E2組和G1組時,我們又發(fā)現(xiàn)collagenⅠ、collagenⅢ、TGF-β1的m RNA表達量明顯降低(P0.05);DMSO組相對于空白對照組無明顯差異(P0.05)。結(jié)論:1.雌激素對于兔耳增生性瘢痕具有促進作用。2.G蛋白偶聯(lián)雌激素受體(GPER)對于兔耳增生性瘢痕具有影響作用,激活GPER能夠促進兔耳增生性瘢痕的形成,拮抗GPER后則具有抑制增生性瘢痕的效果,所以G蛋白偶聯(lián)雌激素受體介導(dǎo)雌激素對于兔耳增生性瘢痕具有促進的作用。
[Abstract]:Purpose 1. The effect of female on hypertrophic scar of rabbit ear was observed and verified by local application of estradiol. 2. 2. To study the effects of G protein-coupled estrogen receptor GPER-mediated rapid non-genomic estrogen effect on hypertrophic scar in rabbit ear and its mechanism. Methods: 36 New Zealand big ear rabbits aged 3 to 6 months, weighing 2.5-3 kg and female, were selected, and 6 round wounds with diameter of 10mm were made on the ventral surface of single lateral ear of rabbits after 7 days of adaptive feeding. The rabbit ear hypertrophic scar model was established by removing the chondroid membrane and reaching the chondrocyte layer with a distance of more than 15 mm. According to the random number table, 36 New Zealand rabbits were divided into 6 groups. Each group (n = 6) is a blank control group (DMSO group) is a solvent group (E _ 2 group) is estrogen group / G _ 1 group) is a GPER agonist group / E _ 2 G15 group (n = 6) estrogen GPER antagonist group / G _ 1 / G _ 1 group) is a GPER agonist antagonist group. On the 15th day after modeling, the 4 groups were injected subcutaneously with a microsyringe to the root of rabbit ear with E2 0.025mg/(kg DX G1 0.06 mg/(kg DU G150.06 mg/(kg dU. The solvent group was injected with 10% DMSO solution diluted by PBS, and the control group was not treated with any treatment. After 14 days of continuous injection, the scar proliferation in each group was observed and the specimens were taken. The expression of TGF- 尾 1 collagen IIIcollagen I was detected by PCR. The result is 1: 1. Morphological observation: compared with the control group, the normal skin around the scar protruding was more obvious, the darker color was dark red, the thickest part was in the central part, and the texture of E2 G15 group and G1 G15 group were stiffer than that of the control group, compared with E2 group and G1 group. The thickness of the center of the scar was obviously reduced, and the color was lighter and pink. Some of the scars were close to normal skin. There was no significant difference in scar proliferation between DMSO group and blank control group. 2. He staining and Masson staining observation showed that there were a large number of fibroblasts in scar tissue and blood vessel growth was abundant in group G 1 and E 2 group compared with control group. A large number of extracellular matrix (ECM) was deposited, and collagen fibers were more and larger. Compared with E2 group and G1 G15 group, fibroblasts were decreased, vascular distribution was less, and extracellular matrix deposition was decreased, compared with E2 group and G1 G15 group. The distribution of fibroblasts and blood vessels in DMSO group was not abnormal compared with that in control group. The scarring hyperplasia index of E2 group and G1 group was significantly higher than that of control group P0.05, E2 G15 group was significantly lower than that of E2 group, and that of G15 group was also significantly lower than that of G1 group, but there was no significant difference between the blank control group and DMSO group. 4. RT-PCR: compared with the blank control group, the ratio of G15 group to the control group was significantly lower than that of the control group. 4. RT-PCR analysis showed that there was no significant difference between the control group and the DMSO group. The mRNA expression of collagen 鈪，

本文編號：2021699