Study On The Relationship Between Seminal Vesicle Inflammation And Sperm Quality

Part I: Construction of rat model of seminal vesiculitis to explore the pathological mechanism of seminal vesiculitisObjective To construct a novel rat model of seminal vesiculitis to provide an effective method for studying the pathogenesis of this disease.Methods Eight male rats received the same operation,during which the root of one of the two seminal vesicleswas partly ligatured with sutures and the other vesicle was left intact.The samples of seminal vesicles were harvested on the 8 thday following the operation.Hematoxylin and eosin and Masson’s trichrome stains were used to observe the histopathology and thepresence of fibrous tissue in seminal vesicles,respectively.Immunoblotting and immunohistochemistry were applied to determine thetumor necrosis factor-alpha and cyclooxygenase-2 levels in seminal vesicle tissues.We applied real-time fluorescence quantitative polymerasechain reactionto detect proinflammatory cytokinesgene expression levels.H 2 O 2 levels in the seminalplasma from the seminal vesicle were also measured.Results Hematoxylin and eosin staining suggested that there was inflammatory cell infiltration into the seminal vesicles treated by partial root ligation.The tumor necrosis factor-alpha and cyclooxygenase-2 proteinswere significantly upregulated in the treated seminal vesicles(P < 0.0001).The tumor necrosis factor-alpha,cyclooxygenase,interleukin 6,andinducible nitric oxide synthase m RNA expression levels were also upregulated in the treated seminal vesicles(P < 0.05).The H 2 O 2 levels inthe seminal plasma from seminal vesicles with partial root ligation were significantly elevated compared with those from vesicleleft intact(P < 0.05).Conclusion Partially ligating the root of the seminal vesicle via sutures in rats is an effective method to establish aseminal vesiculitis rat model.Part II : Comparison of sperm quality related parameters and seminal inflammatory factor levels in normal healthy men and seminal vesicle patientsObjective To study the relationship between seminal vesiculitis and sperm quality parameters and the level of seminal plasma inflammatory mediators.Methods Eighteen patients with persistent blood spermatozoa who underwent transurethral seminal vesicle microscopy and 18 healthy volunteers with normal sperm quality parameters were prospectively collected.Sperm samples were collected from the seminal fluid of patients with persistent blood sperm via transurethral seminal vesicle microscopy.Sperm samples from the control group were collected by masturbation of healthy volunteers.Sperm quality was analyzed under the application of the WLJY-9000 color seminal plasma analysis system.Tumor necrosis factor alpha(TNF-α)and interleukin-6(IL-6)levels of seminal plasma samples were analyzed using an ELISA assay.The level of H2O2 in the seminal plasma was measured using a specific kit.Results The quality of sperm in semen samples from patients with seminal vesicles is significantly lower than that of normal healthy people.Compared with the control group of 11.07%,the mean ratio of the forward + non-forward movement sperm ratio in the seminal vesicle group was 71.54%,and there was a statistically significant difference between the two(P < 0.002).Compared with the control group,TNF-α,IL-6(P < 0.001)and H2O2 levels(P < 0.05)in the seminal vesicle group were significantly increased.Conclusion Seminal vesiculitis is closely related to high levels of inflammatory factors and H2O2 and decreased sperm quality.Part III: Relationship between anti-oxidase Nrf2 and semi-sperm quality associated with seminal vesiculitisObjective To study the relationship between seminal vesiculitis and sperm quality,and related antioxidantfactors.Methods Eighteen patients with persistent blood spermatozoa who underwent transurethral seminal vesicle microscopy and 18 healthy volunteers with normal sperm quality parameters were prospectively collected.For the seminal vesiculitis group,transurethral seminal vesicle microscopy was performed,and semen samples were collected from the seminal vesicle fluid of patients with seminal vesiculitis.For the control group,the sperm samples were collected by masturbation of healthy volunteers.The sperm in the seminal vesicle group and the control group were purified by non-continuous percoll cell separation gradient centrifugation.The m RNA expression of PRM2,TLR2 and TLR4 in the isolated sperm was detected by PCR.Sperm proteins were lysed by cytoplasmic lysate and nuclear lysate,respectively,for related protein detection as H3 and GAPDH.The expression level of sperm Nrf2 protein was detected by immunofluorescence and western blot.Results Compared with normal healthy human semen,the expression level of sperm Nrf2 was significantly increased in the semen samples of patients with persistent blood sperm,and Nrf2 was expressed in sperm cytoplasm and nucleus.Conclusion The sperm ROS level and Nrf2 expression level in semen of patients with seminal vesicles were significantly higher than those in normal healthy people,and the difference was statistically significant(P<0.05).Nrf2 is expressed in the cytoplasm and nucleus of sperm cells.