本文選題：高同型半胱氨酸血癥 + 同型半胱氨酸�。� 參考：《中國人民解放軍軍醫(yī)進修學院》2010年碩士論文

【摘要】： 目的： 1、建立高同型半胱氨酸血癥大鼠模型。 2、采用免疫組化染色和Western blot免疫印跡法研究高同型半胱氨酸血癥大鼠視網(wǎng)膜表達ICAM-1的部位和強度,印證高同型半胱氨酸血癥對視網(wǎng)膜的毒性作用機制。 方法： 1、采用在大鼠每升飲水中添加1.5g L-甲硫氨酸,使大鼠攝入過量甲硫氨酸,飼養(yǎng)大鼠6周,制造高同型半胱氨酸血癥模型。 2、采用HE染色法觀察高同型半胱氨酸大鼠視網(wǎng)膜的結(jié)構(gòu)。 3、應用抗ICAM-1抗體免疫組化方法,檢測大鼠視網(wǎng)膜上ICAM-1的表達量及其表達部位。 4、應用Western blot免疫印跡檢測大鼠視網(wǎng)膜ICAM-1的表達量,印證明免疫組化的檢測結(jié)果。 結(jié)果： 1、采用在大鼠每升飲水中添加1.5g L-甲硫氨酸,飼養(yǎng)大鼠6周后,采大鼠尾靜脈血檢測血漿同型半胱氨酸,對照組血漿Hcy平均濃度為7.0146±2.00μmol／L,模型組為42.9081±12.02μmol／L,對照組中所有大鼠個體血漿Hcy濃度均小于15μmol／L,模型組中所有大鼠個體血漿Hcy濃度均大于15μmol／L,沒有大鼠從實驗中剔除。兩組大鼠血漿Hcy濃度經(jīng)t檢驗,P=0.000,P＜0.05,故兩組大鼠血同型半胱氨酸濃度差別有統(tǒng)計學意義。顯示造模成功。 2、在HE切片上,對照組和模型組大鼠視網(wǎng)膜組織結(jié)構(gòu)未見明顯變化。兩組大鼠視網(wǎng)膜均未見出血、炎性細胞浸潤、血管閉塞等改變。 3、免疫組化檢查發(fā)現(xiàn)對照組OD值為0.284±0.0383；模型組為0.486±0.0329。兩組之間有明顯統(tǒng)計學差異。 4、對照組平均ICAM-1／β-actin比值為0.35±0.04,模型組為0.49±0.03,兩組大鼠視網(wǎng)膜組織Western blot實驗顯色條帶OD值經(jīng)t檢驗,P=0.000,P＜0.05,故兩組OD值差別有統(tǒng)計學意義。 結(jié)論 1、0.15%濃度的甲硫氨酸飲用水能順利誘導出中度升高的高同型半胱氨酸血癥大鼠,顯示動物模型制造成功； 2、用免疫組化和Western bolt免疫印跡法在大鼠視網(wǎng)膜上檢測到了ICAM-1; 3、高同型半胱氨酸血癥能促進大鼠視網(wǎng)膜ICAM-1表達量上升； 4、在參加實驗的所有動物中,未發(fā)現(xiàn)眼底有明顯病變,提示高同型半胱氨酸血癥致病作用的發(fā)生可能還需要更長的時間,或同型半胱氨酸需要聯(lián)合其他致病因素才能起作用。
[Abstract]:Objective: 1. The model of hyperhomocysteinemia was established in rats. 2. The localization and intensity of ICAM-1 expression in the retina of rats with hyperhomocysteinemia were studied by immunohistochemical staining and Western blot immunoblotting to confirm the toxic mechanism of hyperhomocysteinemia on retina. Methods: 1. By adding 1.5 g L-methionine to every liter of drinking water, the rats were fed with excessive methionine for 6 weeks, and the model of hyperhomocysteinemia was established. 2. The structure of high homocysteine rat retina was observed by HE staining. 3. The expression and location of ICAM-1 in rat retina were detected by immunohistochemical method with anti-ICAM-1 antibody. 4. Western blot Western blot was used to detect the expression of ICAM-1 in rat retina. Results: 1. The rats were fed with 1.5 g L-methionine per liter of drinking water for 6 weeks. The plasma homocysteine was detected in the caudal vein blood of the rats. The average plasma Hcy concentration was 7.0146 鹵2.00 渭 mol / L in the control group, 42.9081 鹵12.02 渭 mol / L in the model group, and less than 15 渭 mol / L in all the rats in the control group. The plasma Hcy concentration of all the rats in the model group was greater than 15 渭 mol / L, and no rats were excluded from the experiment. The plasma Hcy concentration of the two groups was tested by t test (P < 0.05), so the difference of plasma homocysteine concentration between the two groups was statistically significant. The model is shown to be successful. 2. In HE section, there was no obvious change in retinal tissue structure in control group and model group. No hemorrhage, inflammatory cell infiltration and vascular occlusion were found in the retina of both groups. 3The OD value of the control group was 0.284 鹵0.0383, and that of the model group was 0.486 鹵0.0329. There was a significant difference between the two groups. 4. The average ratio of ICAM-1/ 尾 -actin was 0.35 鹵0.04 in the control group and 0.49 鹵0.03 in the model group. The OD value of the Western blot in the two groups was significantly higher than that in the control group (P < 0.05). Conclusion (1) 0.15% methionine drinking water could induce moderately elevated hyperhomocysteinemia rats, indicating that the animal model was made successfully. 2ICAM-1 was detected in rat retina by immunohistochemistry and Western bolt immunoblotting. 3. Hyperhomocysteinemia could promote the increase of ICAM-1 expression in rat retina. 4. In all the animals involved in the experiment, there were no obvious lesions in the fundus, suggesting that the pathogenicity of hyperhomocysteinemia may take longer, or that homocysteine needs to be combined with other pathogenic factors to play its role.
【學位授予單位】：中國人民解放軍軍醫(yī)進修學院
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R774.1

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相關(guān)期刊論文 前1條

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