GPER对于兔耳增生性瘢痕作用的研究

发布时间：2018-06-15 10:28

  本文选题：G蛋白偶联雌激素受体 + 增生性瘢痕　； 参考：《遵义医学院》2017年硕士论文

【摘要】：目的:1.通过局部应用雌素来观察并验证其对于兔耳增生性瘢痕的影响作用。2.研究G蛋白偶联雌激素受体(GPER)介导的雌激素快速非基因组效应对于兔耳增生性瘢痕的影响及作用机制。方法:选取3-6月龄新西兰大耳兔,36只,体重2.5-3kg,雌性,要求兔耳皮肤完整无破损;适应性饲养7天后,在兔的单侧耳腹面共做6个直径为10mm的圆形创面,间距大于15mm,深达软骨层并刮除软骨膜,建立兔耳增生性瘢痕模型。按照随机数字表法将36只新西兰大耳兔分为6个组,每组6只:(control组)为空白对照组;(DMSO组)为溶剂组;(E2组)为雌激素组;(G1组)为GPER激动剂组;(E2+G15组)为雌激素+GPER拮抗剂组;(G1+G15组)为GPER激动剂+拮抗剂组。于造模后第15天,4个给药组分别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),溶剂组注射经PBS稀释为10%的DMSO溶液,对照组不予任何处理。持续注射14天后,观察各组瘢痕增生情况并取标本,对比各组瘢痕增生指数(SEI)、成纤维细胞(FB)、胶原纤维以及通过PCR检测TGF-β1、collagen III、collagen I的表达量。结果:1.形态学观察:E2组和G1组相对于空白对照组,瘢痕突出周围正常皮肤更为明显,颜色较深多为暗红色,中央部最为厚实,质地较硬;E2+G15组和G1+G15组对比E2组和G1组,瘢痕中央厚度明显降低,颜色也较浅为粉红色,部分与正常皮肤接近,质地明显变软;DMSO组相对于空白对照组瘢痕增生情况差别不大。2.HE染色和MASSON染色观察:E2组和G1组相对于空白对照组,瘢痕组织中存在大量的成纤维细胞,血管生长丰富,细胞外基质有大量沉积,胶原纤维多而粗大,排列紊乱。E2+G15组和G1+G15组对比E2组和G1组,成纤维细胞明显减少,血管分布较少,细胞外基质沉积减少,胶原纤维也相对减少,DMSO组相对于空白对照组胶原纤维、成纤维细胞和血管分布未见特殊异常。3.瘢痕增生指数:E2组和G1组明显高于空白对照组P0.05),E2+G15组相对于E2组明显降低(P0.05),G1+G15组相对于G1组也明显降低(P0.05),而空白对照组和DMSO组无明显差异(P0.05)。4.RT-PCR检测:与空白对照组相比,E2组和G1组的collagenⅠ、collagenⅢ、TGF-β1的m RNA表达量明显升高(P0.05);而E2+G15组和G1+G15组分别对比E2组和G1组时,我们又发现collagenⅠ、collagenⅢ、TGF-β1的m RNA表达量明显降低(P0.05);DMSO组相对于空白对照组无明显差异(P0.05)。结论:1.雌激素对于兔耳增生性瘢痕具有促进作用。2.G蛋白偶联雌激素受体(GPER)对于兔耳增生性瘢痕具有影响作用,激活GPER能够促进兔耳增生性瘢痕的形成,拮抗GPER后则具有抑制增生性瘢痕的效果,所以G蛋白偶联雌激素受体介导雌激素对于兔耳增生性瘢痕具有促进的作用。
[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，

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