| Objectives:In this study,a rat model of chronic nonbaterial prostatitis(Chronic nonbaterial prostatitis,CNBP)was established based on local injection of Xiaozhiling in the prostate.After 2 weeks of high,low and minimal energy extracorporeal shock wave therapy or Qianliekang intervention,Interleukin 8 and Interleukin 10 in the serum were observed,Ttumor necrosis factor-α,Interleukin 1β,Interferonγ in serum and prostate,P38 protein kinase signal pathway and related downstream signal pathway nuclear tran-κB p65 factor in prostate tissue were also observed.To observe the expression of vascular endothelial growth factor and its receptor 2 in prostate tissue.Thus,the mechanism of the effects of different energy radial extracorporeal shock waves on chronic nonbacterial Prostatitis was investigated based on the p38MAPK/NF-KB signaling pathway.Methods:Sixty Wistar rats(SPF 240-280g)were randomly divided into two groups:Control group(Control group,C group,n=10)were injected with normal saline into the prostate gland,and model group(CNBP group,n=50)were injected with XiaoZhiLing into the prostate gland.The rat model of chronic nonbacterial prostatitis was established in the third week.Rats in the CNBP group were randomly divided into four groups:high energy REWT group(HESWT+CNPH group,n=10),low energy REWT group(LESWT+CNPH group,n=10),minimal energy REWT group(FESWT+CNPH group,n=10),and drug group(P+CNBP group,n=10).The intervention was initiated after one week of adaptive feeding.Rats in the HESWT+CNPH group,LESWT+CNPH group and FESWT+CNPH group were prepared with abdominal skin,coated with coupling agent,and treated with Masterpuls extracorporeal shock wave treatment system(Storz Medical Masterpulsmp 100 barometric pressure wave treatment system,Switzerland).The Ceramic transmitter center was placed at the lower abdominal muscle group of rats,and the shock wave intensity was 3.0bar,1.0bar and 1.0bar(separated by 3cm thick towel),with a frequency of 120 times/min,3Hz,and a total of 300 shock times,once every other day,for 7 times in total.In P+CNBP group,Qianliekang was uncoated and distilled water was added to make suspension of 0.054g/mL.The suspension of Qianliekang was intragastric injected 1 mL/time,5 days a week,for a total of 2 weeks.After 2 weeks of intervention,blood was collected for euthanasia,Prostate tissue was stripped and weighed(PW),and Prostate index(PI)was calculated.Histopathological changes of prostate were observed by HE staining.Enzyme-linked immunosorbent assay(ELISA)was used to determine the levels of serum IL-8 and IL-10,and the levels of TNF-α,IL-1β and IFN-y in prostate homogenate tissue.The expressions of VEGFR2,NF-kB p65,p38 MAPK and IKB-α in prostate tissues were detected by immunohistochemistry(IHC).The relative expression levels of VEGF,NF-kB p65,p38 MAPK and IKB-α in prostate tissues were detected by Western blotting(WB).The experimental results were expressed as mean plus or minus standard deviation(M±SD).The differences between the two groups were tested by independent sample t test.One-way ANOVA was used to analyze the differences between the groups.p<0.05 was used for significance.Results:(1)HE staining results showed that the prostate gland cavity of rats in group C was normal and the cell morphology was normal and regular.Compared with group C,the glandular lumen of rats in CNBP group was damaged and a large number of inflammatory cells were infiltrated in the glandular stroma,indicating that the model of CNBP rats was successfully constructed.Compared with the CNBP group,the damage of the prostatic gland cavity of rats in the HESWT+CNBP group,the LESWT+CNBP group,the FESWT+CNBP group and the P+CNBP group were improved and the infiltration of inflammatory cells was reduced.(2)After 2 weeks of intervention,there were no significant differences in PW and PI indexes among groups C,CNBP,HESWT+CNBP,LESWT+CNBP,FESWT+CNBP and P+CNBP(p>0.05).(3)Compared with group C,the contents of IL-8 in serum and TNF-α,IL-1β and IFN-y in prostate tissue of rats in CNBP group were significantly increased(p<0.05),and the serum IL-10 content was significantly decreased(p<0.05).Compared with CNBP group,the contents of IL-8,TNF-α,IL-1β and IFN-y in HESWT+CNP group,LESWT+CNP group,FESWT+CNBP group and P+CNBP group were significantly decreased,while the content of IL-10 was significantly increased.The results showed significant difference between the two groups(p<0.05).The contents of IL-8,TNF-α,IL-1β and IFN-y in P+CNBP group were significantly lower than those in HESWT+CNBP group,while the contents of IL-10 were significantly higher(p<0.05).The contents of IL-8,TNF-α,IL-1β and IFN-y in HESWT+CNBP group were significantly lower than those in FESWT+CNBP group,while the content of IL-10 was significantly higher(p<0.05).Compared with LESWT+CNBP group,the contents of IL-8,TNF-α,IL-1β and IFN-y in FESWT+CNBP group were significantly decreased,while the content of IL-10 was significantly increased(p<0.05).(4)IHC results showed that the expressions of VEGFR2,NF-kB p65 and p38 MAPK in the prostate tissues of rats in CNBP group were significantly higher than those in C group by immunohistochemical staining and analysis of the Average Optical Density(AOD)of each group(p<0.05),the positive expression of IKB-α was significantly decreased(p<0.05).Compared with CNBP group,VEGFR2,NF-kB p65,p38 MAPK were significantly decreased in HESWT+CNBP group and P+CNBP group(p<0.05),the positive expression of IKB-α was significantly increased(p<0.05),the p38 MAPK of FESWT+CNBP group was significantly decreased(p<0.05),the positive expression of IKB-α was significantly increased(p<0.05),there was no significant difference in the positive expression of NF-KB p65,IKB-α,p38 MAPK and VEGFR2 in LESWT+CNP group(p>0.05).(5)WB results showed that compared with the data in group C,the total protein levels of VEGF,NF-KB p65 and P38 MAPK in the prostate tissue of rats in CNBP group were significantly increased(p<0.05),the total protein content of IKB-α was significantly decreased(p<0.05).Compared with CNBP group,the total protein expressions of VEGF,NF-KB p65 and P38 MAPK in HESWT+CNBP group and P+CNBP group were down-regulated(p<0.05),the total protein content of IKB-α was significantly increased(p<0.05).Compared with the CNBP group,there was no statistical difference in rats in the LESWT+CNBP group and the FESWT+CNBP group(p>0.05).There was no significant difference in total NF-KB p65 protein between P+ESWT group and HESWT+CNBP group(p>0.05).Conclusion(1)Xiaozhiling local injection of prostate successfully induced chronic non-bacterial prostatitis in rats,and maintained for more than 6 weeks,and a rat CNBP model was successfully established.(2)Two weeks high,low,minimal energy spread of extracorporeal shock wave intervention can reduce CNBP rats serum IL-8 levels,lower prostate tissue and serum IL-1β,IFN-α and TNF-α level,increase the serum level of IL-10,significantly improve the inflammation of the prostate tissue of rats,including high-energy extracorporeal shock wave therapy is superior to the low,minimal energy,Qianliekang is superior to high-energy extracorporeal shock wave therapy.(3)The mechanism of high energy extracorporeal shock wave therapy and Qianliekang drug therapy on chronic nonbaterial prostatitis in rats may be through down-regulating the expression of NF-KB P65,P38 MAPK and their related proteins in Prostatitis tissues,down-regulation of the expression of VEGF and VEGF receptor 2 is achieved,thus improving the microcirculation of prostate tissue,to achieve the goal of treatment of CNBP(4)Two weeks of radial extracorporeal shock wave therapy or drug treatment had no significant effect on the body weight and prostate index of rats. |
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