| Objective: Aim to investigate the significance of expression of Mast Cells ,NGF and ASIC3 in prostate of experimental rats with type Ⅲ prostatitis.Methods: Randomly allocation was taken to averagely divide 60 male SD rats into four groups:Control group(15 rats)(Pelvic area and bilateral scapular subcutaneous injection of 0.9% sodium chloride),Experimental Autoimmune Prostatitis model group/EAP group(15 rats)(Pelvic area and bilateral scapular subcutaneous injection of the mixed suspension of Complete Freund’s adjuvant and prostate tissue).Intervention group(Ⅰ)(EAP rats treated with cromolyn sodium).Intervention group(Ⅱ)(EAP rats treated with amiloride).Tactile allodynia was quantified using Von-Frey filaments as a measure of pelvic pain behavior.And Von-Frey filaments was performed on the 5th,10th,20th,30th and 40th day in controlgroup and EAP group.When the EAP rat model was established successfully,the rats in Intervention group(Ⅰ) and Intervention group(Ⅱ) were given drugs for 10 days respectively, and during this time we tested the tactile allodynia in Intervention group(Ⅰ),Intervention group(Ⅱ) and EAP group every other days.Mouse prostate were collected from rats and were processed for H&E staining and and toluidine blue staining,while the expressions of mast cell tryptase,NGF and ASIC3 were measured by Immunohistochemical staining and Western blot.Results: 1.Von-Frey test results shown that the pelvis pain was significantly more obvious in EAP model group than that in control group.However,the pelvic pain in EAP model group was lighter than that in EAP group.HE staining shown that the lymphocytes and neutrophils infiltrated in the prostate of EAP rats,and the surrounding glands were congested.No inflammatory cell was founded in the control group. 2.In the study of mast cells in rat prostate,toluidine blue staining shown that the number of mast cells and degranulated mast cells are increased in EAP group compared with those in control group(P<0.01),and decreased’ in EAP group(P<0.05).HE staining shown that the congestion of the gland and the infiltration of the inflammatory cells were alleviated,when intervention group( I) compared with EAP group.Immunohistochemical staining results shown that the expression of mast cells tryptase was significantly increased(P<0.01) in EAP group when compared with control group,while in intervention group( I ) shown reduced expression(P<0.05).Western blot results shown that the mast cells tryptase was expressed higher in EAP group when compared with control group(P<0.01),while in intervention group shown reduced expression.Von-Frey test results shown that the pelvis pain in Intervention group( I ) was lighter than that in EAP group(P<0.05). 3. In the study of NGF in rat prostate,immunohistochemical staining results shown that the expression of NGF was significantly increased(P<0.01) in EAP group when compared with control group. Western blot results shown that the NGF were expressed higher in EAP group when compared with control group(P<0.01). 4.In the study of ASIC3 in rat prostate,HE staining shown that,the congestion of the gland and the infiltration of the inflammatory cells were not alleviated,when intervention group( II) compared with EAP group.Immunohistochemical staining results shown that the expression of ASIC3 was significantly increased(P<0.01) in EAP group when compared with control group,while in intervention group shown reduced expression(P<0.05).Western blot results shown that ASIC3 was expressed higher in EAP group when compared with control group(P<0.0 1),while in intervention group shown reduced expression.Von-Frey test results shown that the pelvis pain in Intervention group( II)was lighter than that in EAP group(P<0.05).Conclusion(s): 1.In this study,we used the autoimmune method of multi-point injection of the mixed suspension of Complete Freund’s adjuvant and prostate tissue adjuvant to established the experimental model of type Ⅲ prostatitis successfully.2.Rats in the control group, EAP group and intervention group (Ⅰ) were quantified for pelvic pain by using the Von-Frey fiber,the results are complaince with the chang of HE stanning in prostate of rats. 3.The number of mast cells and degranulated mast cells in prostate of EAP rats are significantly increased. Mast cells are one of the most important mediators of chronic pelvic pain caused by type Ⅲ prostatitis,which can be used as a target for therapeutic intervention and treatment in type Ⅲprostatitis. 4.The expression of NGF in prostate of EAP rats was increased,and NGF was positively correlated with chronic pelvic pain in EAP rats. 5.The expression of ASIC3 in prostate of EAP rats was increased significantly. ASIC3 are one of the most important mediators of chronic pelvic pain caused by type Ⅲ prostatitis,which can be used as a target for therapeutic intervention and treatment in type Ⅲ prostatitis. |
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