本文關(guān)鍵詞：重組人促紅細胞生成素對眼挫傷后大鼠視網(wǎng)膜保護作用機制　出處：《遼寧醫(yī)學(xué)院》2012年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 重組人促紅細胞生成素 大鼠 眼挫傷 B細胞淋巴瘤/白血病基因2 BCL-2相關(guān)X蛋白

【摘要】：目的 通過建造大鼠視網(wǎng)膜挫傷模型，觀察視網(wǎng)膜內(nèi)Bcl-2和Bax蛋白表達的變化，探討重組人促紅細胞生成素對挫傷視網(wǎng)膜Bcl2和Bax的調(diào)節(jié)作用，明確重組人促紅細胞生成素是否對挫傷的大鼠視網(wǎng)膜具有保護作用。探討重組人促紅細胞生成素對眼挫傷大鼠視網(wǎng)膜保護作用的可能機制。 方法 雄性大鼠45只，兩月齡，5只用于正常對照組，余下40只仿Allen重擊法制作視網(wǎng)膜挫傷模型致左眼挫傷。將模型制備成功大鼠隨機分為兩組：模型組和rhEPO治療組。每組又分12h，24h，3d，7d四個時間點，每個時間點大鼠5只。rhEPO組腹腔注射rhEPO（60IU/Kg），每日一次；模型組用同體積的生理鹽水腹腔注射。各時段于給藥給水停止后第2天取材，石蠟切片。應(yīng)用HE染色方法觀察各組視網(wǎng)膜各層細胞的形態(tài)改變；應(yīng)用免疫組織化學(xué)方法檢測視網(wǎng)膜組織中Bcl-2和Bax蛋白的表達變化。 結(jié)果 1、視網(wǎng)膜各層組織病理學(xué)改變 HE染色觀察可見正常對照組視網(wǎng)膜各層細胞數(shù)量多，排列整齊，形態(tài)較完整。而模型組與rhEPO組視網(wǎng)膜全層細胞數(shù)減少，視網(wǎng)膜厚度較正常組明顯變薄，各層細胞排列紊亂，細胞水腫。 2、Bcl-2蛋白檢測結(jié)果 Bcl-2免疫組織化學(xué)染色顯示：光鏡下Bcl-2免疫反應(yīng)產(chǎn)物呈棕黃色。正常眼視網(wǎng)膜Bcl-2蛋白在神經(jīng)節(jié)細胞層、內(nèi)核層和外核層細胞的胞漿均有表達。視網(wǎng)膜挫傷后，Bcl-2蛋白表達隨時間的延長逐漸下降，至傷后3d，下降到最低點，7d回升。在模型組，除傷后12h外，其余時間點Bcl-2蛋白表達明顯低于正常組（P0.05）；在rhEPO組，，Bcl-2蛋白表達變化趨勢與模型組相似，但在24h、3d、7d，Bcl-2蛋白表達較模型組明顯增強，其蛋白陽性產(chǎn)物的平均光密度值(Mean optical density,MOD)明顯高于模型組（P0.01）。各組大鼠的陰性對照片未見陽性反應(yīng)發(fā)生。 3、Bax蛋白檢測結(jié)果 光鏡下，Bax蛋白免疫反應(yīng)產(chǎn)物呈棕黃色。正常組視網(wǎng)膜Bax蛋白在神經(jīng)節(jié)細胞層、內(nèi)核層和外核層及感光細胞層的胞漿均有表達。挫傷后12h，視網(wǎng)膜Bax蛋白的表達與正常組無顯著性差異（P0.05）。但挫傷后24h的表達明顯增加，3d達最高峰，7d回落。除傷后12h外，模型組Bax蛋白表達比正常對照組顯著增高（P0.01）。在rhEPO組，除12h外，Bax蛋白表達在其余時間點較模型組明顯降低，其蛋白陽性產(chǎn)物的平均光密度值(Mean optical density, MOD)明顯低于模型組（P0.01）。 結(jié)論 Bcl-2和Bax參與了眼挫傷視網(wǎng)膜病變過程；rhEPO能夠調(diào)節(jié)挫傷后大鼠視網(wǎng)膜細胞凋亡因子Bcl-2和Bax的表達，抑制視網(wǎng)膜損傷后神經(jīng)元的凋亡。rhEPO對眼挫傷視網(wǎng)膜病變有保護作用。
[Abstract]:Purpose The changes of Bcl-2 and Bax protein expression in retina were observed and the regulatory effects of recombinant human erythropoietin on Bcl2 and Bax in retina contusion were studied by establishing a rat model of retinal contusion. To determine whether recombinant human erythropoietin has protective effect on the retina of contusion rats and to explore the possible mechanism of the protective effect of recombinant human erythropoietin on the retina of eye contusion rats. Method 45 male rats and 5 male rats of two months old were used as normal control group. The other 40 rats were randomly divided into two groups: model group and rhEPO group, each group was divided into 12 hours and 24 hours. Five rats in each time point were injected intraperitoneally with rhEPO-60IUP / KG once a day. The rats in the model group were injected intraperitoneally with normal saline of the same volume. The samples were taken on the second day after the administration of water and the paraffin sections were taken. The morphological changes of the cells in each layer of retina were observed by HE staining. Immunohistochemical method was used to detect the expression of Bcl-2 and Bax in retinal tissues. Results 1. Histopathological changes in all layers of retina He staining showed that the number of retinal cells in the normal control group was more than that in the control group, and the number of cells in the whole retinal layer was decreased in the model group and rhEPO group. The thickness of retina was obviously thinner than that of normal group, the cells in each layer were disordered, and the cells were edema. 2detection of Bcl-2 protein Bcl-2 immunohistochemical staining showed that the immunoreactive product of Bcl-2 was brown under light microscope, and Bcl-2 protein of retina of normal eyes was in ganglion cell layer. After retinal contusion, the expression of Bcl 2 protein decreased gradually with the extension of time, then decreased to the lowest point at 3 days after injury, and increased at the lowest point 7 days after injury. In the model group, the expression of Bcl 2 protein decreased gradually after retinal contusion. The expression of Bcl-2 protein was significantly lower than that of normal group at 12 h after injury. The change trend of Bcl-2 protein expression in rhEPO group was similar to that in model group, but the expression of Bcl-2 protein was significantly increased at 24 h, 3 d, 7 d, compared with model group. The average optical density of its protein positive product was mean optical density. MOD was significantly higher than that in the model group (P 0.01). No positive reaction was found in the negative photos of the rats in each group. Detection of Bax protein The immunoreactive product of Bax protein was brown under light microscope. Bax protein was expressed in the cytoplasm of ganglion cell layer, nuclear layer, outer nuclear layer and photosensitive cell layer in normal group. 12 hours after contusion, Bax protein was expressed in the cytoplasm of the ganglion cell layer, the inner and outer nuclear layer and the photosensitive cell layer. The expression of Bax protein in the retina was not significantly different from that in the normal group (P 0.05). However, the expression of Bax protein increased significantly at 24 h after contusion and reached the peak on the 3rd day and decreased on the 7th day, except for 12 h after injury. The expression of Bax protein in the model group was significantly higher than that in the normal control group (P 0.01). The expression of Bax protein in the rhEPO group was significantly lower than that in the model group at other time points except 12 h. The mean optical density (mod) of the protein positive product was significantly lower than that of the model group (P 0.01). Conclusion Bcl-2 and Bax were involved in the process of ocular contusion retinopathy. RhEPO can regulate the expression of Bcl-2 and Bax in rat retinal cells after contusion. Inhibition of neuronal apoptosis after retinal injury. RhEPO has protective effect on retinal contusion.
【學(xué)位授予單位】：遼寧醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R779.1

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