【摘要】：[目的]探討前列腺組織中肥大細(xì)胞、NGF及ASIC3在實驗性大鼠Ⅲ型前列腺炎慢性骨盆疼痛中的表達(dá)的意義。[方法]選取雄性SD大鼠60只,分為對照組15只(0.9%NaCl盆腔區(qū)域皮下、雙側(cè)肩胛皮下多點注射)、Ⅲ型前列腺炎模型/EAP組15只(完全弗氏佐劑與前列腺組織混合懸液盆腔區(qū)域皮下、雙側(cè)肩胛皮下多點注射)、干預(yù)組(Ⅰ )15只(EAP模型組基礎(chǔ)上加入肥大細(xì)胞類胰蛋白酶脫落抑制劑色甘酸鈉),干預(yù)組(Ⅱ) 15只(EAP模型組基礎(chǔ)上加入ASIC3抑制劑阿米洛利)。建立EAP模型過程中,予Von-Frey測痛纖維于第0d、5d、10d、20d、30d、40d于對照組、EAP組進(jìn)行骨盆區(qū)域痛覺測試。建模成功后,干預(yù)組(Ⅰ)予色甘酸鈉連續(xù)腹腔內(nèi)注射10d,干預(yù)組(Ⅱ)予阿米洛利連續(xù)腹腔內(nèi)注射10d,每2d對干預(yù)組(!)、干預(yù)組(Ⅱ)及EAP組進(jìn)行痛覺測試。選取4組大鼠的前列腺,進(jìn)行HE染色及甲苯胺藍(lán)染色病理學(xué)檢查,并通過免疫組化和Western blot方法檢測肥大細(xì)胞類胰蛋白酶、NGF及ASIC3的表達(dá)。[結(jié)果]1.Von-Frey測痛結(jié)果顯示EAP組大鼠骨盆疼痛表現(xiàn)較對照組明顯升高(P 0.01)。HE染色:EAP組大鼠前列腺呈不規(guī)則改變,腺體間質(zhì)內(nèi)淋巴細(xì)胞和中性粒細(xì)胞浸潤,腺體周圍充血;對照組大鼠前列腺未見明顯炎癥細(xì)胞浸潤及充血。2.大鼠前列腺組織肥大細(xì)胞的研究中,甲苯胺藍(lán)染色:肥大細(xì)胞總數(shù)及脫顆粒的肥大細(xì)胞數(shù)目較對照組增加(P 0.01),干預(yù)組(Ⅰ)較EAP組減少(P 0.05)。HE染色:干預(yù)組(Ⅰ )腺體充血及炎癥細(xì)胞浸潤情況較EAP組減輕。免疫組化染色提示:肥大細(xì)胞類胰蛋白酶較對照組表達(dá)增加(P0.01),干預(yù)組(Ⅰ)較EAP組減少(P 0.05); Western blot提示:肥大細(xì)胞類胰蛋白酶較對照組表達(dá)增加(P0.01),干預(yù)組較EAP組減少(P 0.01)。Von-Frey測痛結(jié)果顯示干預(yù)組(Ⅰ)大鼠骨盆疼痛表現(xiàn)較EAP模型組減輕(P 0.05)。3.大鼠前列腺組織NGF的研究中,免疫組化提示:NGF的蛋白表達(dá)較對照組表達(dá)增加(P 0.01); Western blot提示:NGF較對照組表達(dá)增加(P 0.01)。4.大鼠前列腺組織ASIC3的研究中,HE染色:干預(yù)組(Ⅱ)腺體充血及炎癥細(xì)胞浸潤情況較EAP組無明顯改變。免疫組化染色提示:ASIC3表達(dá)較對照組表達(dá)增加(P0.01),干預(yù)組(Ⅱ)較 EAP 組減少(P0.05);Western blot 提示:ASIC3較對照組表達(dá)增加(P0.01),干預(yù)組較EAP組減少(P0.01)。Von-Frey測痛結(jié)果顯示干預(yù)組(Ⅱ)大鼠骨盆疼痛表現(xiàn)較EAP模型組減輕(P 0.05)。[結(jié)論]1.采用多點多次皮下注射完全弗氏佐劑與前列腺組織混合懸液的自身免疫法可以成功構(gòu)建實驗性Ⅲ型前列腺炎大鼠模型。2. Von-Frey測痛纖維對對照組、EAP組、干預(yù)組(Ⅰ )進(jìn)行骨盆區(qū)域痛覺測試;測試結(jié)果與各組前列腺的病理改變相吻合。3. EAP大鼠前列腺組織中肥大細(xì)胞的總數(shù)及脫顆粒的肥大細(xì)胞數(shù)目增加,肥大細(xì)胞脫顆粒抑制劑色甘酸鈉可一定程度上可緩解慢性骨盆疼痛癥狀,肥大細(xì)胞是導(dǎo)致Ⅲ型前列腺炎骨盆區(qū)域疼痛介質(zhì)之一。4. EAP大鼠前列腺組織中NGF表達(dá)增加,NGF與EAP大鼠慢性骨盆疼痛呈正相關(guān)。5. EAP大鼠前列腺組織中ASIC3表達(dá)增加,ASIC3抑制劑阿米洛利可一定程度上可緩解慢性骨盆疼痛癥狀,ASIC3是導(dǎo)致Ⅲ型前列腺炎骨盆區(qū)域疼痛介質(zhì)之一。
[Abstract]:[Objective] to investigate the significance of the expression of mast cells, NGF and ASIC3 in the chronic pelvic pain of experimental rat type III prostatitis in the prostate tissue. [Methods] 60 male SD rats were selected to be divided into 15 control groups (0.9%NaCl pelvic region subcutaneous, bilateral subcutaneous injection of scapula), and 15 (complete Freund) model group III prostatitis model group /EAP. The intervention group (I) 15 (EAP model group was added to the mast cell tryptase shedding inhibitor sodium tryptase), and the intervention group (II) 15 (ASIC3 inhibitor amiloride) in the intervention group (EAP model group on the basis of ASIC3 inhibitor amiloride). In the process of establishing the EAP model, Von-Fre was given to the group. Y pain fiber was measured in 0d, 5D, 10d, 20d, 30d, 40d in the control group and EAP group for the pelvic region pain test. After the successful modeling, the intervention group was given a continuous intraperitoneal injection of 10d in the intervention group (II), the intervention group was given a continuous intraperitoneal injection of amiloride, the intervention group (!), the intervention group (II) and the EAP group were tested for the pain, and the 4 groups of rats were selected. The adenoid, HE staining and toluidine blue staining pathology examination, and the immunohistochemistry and Western blot method to detect the expression of mast cell tryptase, NGF and ASIC3. [results]1.Von-Frey test results showed that the pelvic pain in the EAP group was significantly higher than that in the control group (P 0.01).HE staining: the prostate of the EAP group was irregular. Changes in the infiltration of lymphocytes and neutrophils in the glandular mesenchyme and hyperemia around the gland; in the control group, no obvious inflammatory cell infiltration and hyperemia of the prostatic mast cells in.2. rats were observed. Toluidine blue staining: the total number of mast cells and the number of degranulation mast cells increased (P 0.01) in the control group (I) Compared with group EAP (P 0.05).HE staining, the interference group (I) the gland congestion and infiltration of inflammatory cells was less than that of the EAP group. Immunohistochemical staining suggested that the expression of mast cell tryptase increased in comparison with the control group (P0.01), the intervention group (I) decreased (P 0.05) than that of the EAP group (P 0.05); Western blot suggested that the expression of mast cell tryptase was increased compared with the control group (P0). .01), the intervention group was less than the EAP group (P 0.01).Von-Frey pain test results showed that the pelvic pain in the intervention group was better than that of the EAP model group (P 0.05).3. rats. The immunohistochemical staining suggested that the expression of NGF was higher than that of the control group (P 0.01), and Western blot suggested that the expression was increased compared with the control group (0.01). In the study of the ASIC3 of the rat prostate tissue, HE staining: the interference group (II) no obvious changes in the gland congestion and inflammatory cell infiltration in the EAP group. The immunohistochemical staining suggested that the expression of ASIC3 was increased compared with the control group (P0.01), the intervention group (II) was less than that of the EAP group (P0.05); Western blot suggested that the expression of ASIC3 was higher than the control group (P0.01), intervention group (P0.01), intervention group The results of.Von-Frey pain reduction in group EAP (P0.01) showed that the pelvic pain in the intervention group (II) was less than that of the EAP model group (P 0.05). [conclusion the autoimmunity method of multiple subcutaneous injection of complete Freund's adjuvant and the mixed suspension of the prostate tissue by multiple subcutaneous injection of]1. can successfully construct the.2. Von-Frey test for the experimental model of type III prostatitis rats. In the control group, the EAP group and the intervention group (I), the pain in the pelvic region was tested. The results were in agreement with the pathological changes of the prostate in each group. The total number of mast cells in the prostate tissue of.3. EAP rats and the number of degranulation mast cells increased, and the degranulation of mast cell degranulation sodium glycolate could relieve the chronic pelvis to a certain extent. The symptoms of pain, mast cell is one of the pain mediators in the pelvic region of type III prostatitis, one of the.4. EAP rats, the expression of NGF in the prostate tissue is increased. NGF and EAP rat chronic pelvic pain is positively related to the increase of ASIC3 expression in the prostate tissue of.5. EAP rats. ASIC3 inhibitor amiloride can alleviate chronic pelvic pain to some extent. ASIC3 is one of the mediators of pelvic pain in type III prostatitis.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R697.3;R-332

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