肥大细胞、NGF及ASIC3与实验性大鼠Ⅲ型前列腺炎模型慢性骨盆疼痛的相关研究
发布时间：2018-08-06 20:32
【摘要】：[目的]探讨前列腺组织中肥大细胞、NGF及ASIC3在实验性大鼠Ⅲ型前列腺炎慢性骨盆疼痛中的表达的意义。[方法]选取雄性SD大鼠60只,分为对照组15只(0.9%NaCl盆腔区域皮下、双侧肩胛皮下多点注射)、Ⅲ型前列腺炎模型/EAP组15只(完全弗氏佐剂与前列腺组织混合悬液盆腔区域皮下、双侧肩胛皮下多点注射)、干预组(Ⅰ )15只(EAP模型组基础上加入肥大细胞类胰蛋白酶脱落抑制剂色甘酸钠),干预组(Ⅱ) 15只(EAP模型组基础上加入ASIC3抑制剂阿米洛利)。建立EAP模型过程中,予Von-Frey测痛纤维于第0d、5d、10d、20d、30d、40d于对照组、EAP组进行骨盆区域痛觉测试。建模成功后,干预组(Ⅰ)予色甘酸钠连续腹腔内注射10d,干预组(Ⅱ)予阿米洛利连续腹腔内注射10d,每2d对干预组(!)、干预组(Ⅱ)及EAP组进行痛觉测试。选取4组大鼠的前列腺,进行HE染色及甲苯胺蓝染色病理学检查,并通过免疫组化和Western blot方法检测肥大细胞类胰蛋白酶、NGF及ASIC3的表达。[结果]1.Von-Frey测痛结果显示EAP组大鼠骨盆疼痛表现较对照组明显升高(P 0.01)。HE染色:EAP组大鼠前列腺呈不规则改变,腺体间质内淋巴细胞和中性粒细胞浸润,腺体周围充血;对照组大鼠前列腺未见明显炎症细胞浸润及充血。2.大鼠前列腺组织肥大细胞的研究中,甲苯胺蓝染色:肥大细胞总数及脱颗粒的肥大细胞数目较对照组增加(P 0.01),干预组(Ⅰ)较EAP组减少(P 0.05)。HE染色:干预组(Ⅰ )腺体充血及炎症细胞浸润情况较EAP组减轻。免疫组化染色提示:肥大细胞类胰蛋白酶较对照组表达增加(P0.01),干预组(Ⅰ)较EAP组减少(P 0.05); Western blot提示:肥大细胞类胰蛋白酶较对照组表达增加(P0.01),干预组较EAP组减少(P 0.01)。Von-Frey测痛结果显示干预组(Ⅰ)大鼠骨盆疼痛表现较EAP模型组减轻(P 0.05)。3.大鼠前列腺组织NGF的研究中,免疫组化提示:NGF的蛋白表达较对照组表达增加(P 0.01); Western blot提示:NGF较对照组表达增加(P 0.01)。4.大鼠前列腺组织ASIC3的研究中,HE染色:干预组(Ⅱ)腺体充血及炎症细胞浸润情况较EAP组无明显改变。免疫组化染色提示:ASIC3表达较对照组表达增加(P0.01),干预组(Ⅱ)较 EAP 组减少(P0.05);Western blot 提示:ASIC3较对照组表达增加(P0.01),干预组较EAP组减少(P0.01)。Von-Frey测痛结果显示干预组(Ⅱ)大鼠骨盆疼痛表现较EAP模型组减轻(P 0.05)。[结论]1.采用多点多次皮下注射完全弗氏佐剂与前列腺组织混合悬液的自身免疫法可以成功构建实验性Ⅲ型前列腺炎大鼠模型。2. Von-Frey测痛纤维对对照组、EAP组、干预组(Ⅰ )进行骨盆区域痛觉测试;测试结果与各组前列腺的病理改变相吻合。3. EAP大鼠前列腺组织中肥大细胞的总数及脱颗粒的肥大细胞数目增加,肥大细胞脱颗粒抑制剂色甘酸钠可一定程度上可缓解慢性骨盆疼痛症状,肥大细胞是导致Ⅲ型前列腺炎骨盆区域疼痛介质之一。4. EAP大鼠前列腺组织中NGF表达增加,NGF与EAP大鼠慢性骨盆疼痛呈正相关。5. EAP大鼠前列腺组织中ASIC3表达增加,ASIC3抑制剂阿米洛利可一定程度上可缓解慢性骨盆疼痛症状,ASIC3是导致Ⅲ型前列腺炎骨盆区域疼痛介质之一。
[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.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R697.3;R-332
本文编号：2168907
[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.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R697.3;R-332
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