Study On The Intervention Effect Of Qianlie Biertong Suppository On Chronic Nonbacterial Prostatitis And Its Anti-inflammatory Mechanis

Chronic prostatitis is one of the common multiple diseases in male patients,accounting for about 30%of urology diseases.The results of epidemiological studies show that the age of onset of chronic prostatitis is mainly 20 to 55 years old,and its incidence rate is increasing year by year.Chronic nonbacterial prostatitis(CNP)accounts for more than 90%of the incidence rate of clinical prostatitis.The clinical manifestations of CNP patients mainly include various urinary abnormalities,pelvic pain,erectile dysfunction or sexual dysfunction,which can easily cause negative psychological emotions such as anxiety and depression,seriously affecting the patient’s quality of life.For the treatment of this disease,the "Chinese Guidelines for the Diagnosis and Treatment of Urology and Andrology Diseases"(2019 edition)recommends the use of plant preparations and traditional Chinese medicine for the treatment of chronic prostatitis.Therefore,the development of safe and effective traditional Chinese medicine preparations has become particularly important.Chapter 1:Study on the Interventional Effect of Qianlie-Biertong Suppository on Chronic Nonbacterial Prostatitis Rats1.Study on the Interventional Effect of Qianlie-Biertong Suppository on Chemical Prostatitis RatsObjective:To investigate the therapeutic effect of Qianlie-Biertong Suppository on chemical prostatitis in rats.Methods:SD rats were randomly divided into blank group,model group,sham operation group,Qianlie-tong Suppository group,and Qianlie-Biertong Suppository High,Middle,and Low dose groups.Both the model group and the administration group were injected with 0.1 mL of complete Freund’s adjuvant once.After one day of modeling,rectal administration was conducted for 23 days(0.33,0.66,and 0.99 g/d for low,middle,and high dose groups of Qianlie-Biertong suppository,respectively,and 0.42 g/d for Qianlie-tong suppository group).From the prostate index,HE staining pathology,bladder internal pressure change rate,lecithin corpuscles,white blood cell count,tumor necrosis factor-α(TNF-α)、interleukin-10(IL-10),superoxide dismutase(SOD),interleukin-8(IL-8),and malondialdehyde(MDA)content were analyzed.Results:The middle dose of Qianlie-Biertong Suppository reduced the prostate index(compared to the model group,P<0.05).The middle and high doses of Qianlie-Biertong Suppository increased the number of lecithin bodies(compared to the model group,P<0.05).Each dose of Qianlie-Biertong Suppository reduced the number of white blood cells and pathological score(compared with the model group,P<0.05).The rate of changes in bladder internal pressure was increased by various doses of Qianlie-Biertong Suppository(compared with the model group,P<0.05).High dose of Qianlie-Biertong Suppository reduces the levels of IL-8 and MDA in prostate tissue.Each dose of Qianlie-Biertong Suppository increased the IL-10 content in prostate tissue(compared to the model group,P<0.05).The middle dose of Qianlie-Biertong Suppository increased the SOD content in prostate tissue(compared to the model group,P<0.05).Conclusion:Qianlie-Biertong Suppository has a therapeutic effect on chemical prostatitis model rats.2.Study on the therapeutic effect of Qianlie-Biertong Suppository on experimental autoimmune prostatitis in ratsObjective:To investigate the therapeutic effect of Qianlie-Biertong Suppository on experimental autoimmune prostatitis(EAP)in rats.Methods:SD rats were used to establish an EAP rat model using prostate protein purification solution combined with complete Freund’s adjuvant.The other 10 rats were used as a blank group.Fifty EAP rats that successfully established the model were randomly divided into model group,low,middle,and high dose groups of Qianlie-Biertong Suppository,and Qianlie-tong Suppository group.After 28 days of rectal administration,the rate of changes in bladder internal pressure was measured,the number of lecithin bodies and white blood cells were measured,and the inflammatory factors in prostate tissue such as IL-8,IL-10,and TNF-α were detected by ELISA,HE staining was used to observe the pathological changes of prostate tissue,and Western blot was used to detect nuclear factors κB(NF-κB)Subunits p65 in prostate tissue,IκB kinase α(IKK-α)、p-IKK-α、NF-κB inhibitory protein α(κB-α)、p-IκB-α、TNF-α、Cyclooxygenase2(COX2)protein expression,quantitative reverse transcription PCR(qRTPCR)was used to detect chemokine 5(CXCL5),interleukin-6(IL-6),COX-2 and TNF-A gene expression.Results:Each dose of Qianlie-Biertong Suppository can increase the number of lecithin bodies in prostatic fluid of model animals,reduce the number of white blood cells,and alleviate the damage of prostate tissue(compared with the model group,P<0.05).The rate of changes in bladder internal pressure in model animals increased with the middle dose of Qianlie-Biertong Suppository(compared to the model group,P<0.05).Middle and high doses of QianlieBiertong Suppository can reduce in prostate tissue TNF-α,IL-8 content(compared to the model group,P<0.05).Various doses of Qianlie-Biertong Suppository can cause in prostate tissue the expression of p-IκB-α,COX-2 protein decreased(compared to the model group,P<0.05);Low dose of Qianlie-Biertong Suppository can reduce in prostate tissue of model animals pIKK-a protein expression(compared to the model group,P<0.05);Low and high doses of Qianlie-Biertong Suppository can reduce in prostate tissue NF-κB p65 protein expression(compared to the model group,P<0.05);Middle dose of Qianlie-Biertong Suppository can reduce in prostate tissue TNF-α Protein expression(compared to the model group,P<0.05).Various doses of Qianlie-Biertong Suppository can reduce in prostate tissue TNF-A,CXCL5,IL-6,COX-2 gene expression(compared to the model group,P<0.05).Conclusion:Qianlie-Biertong Suppository Suppress NF-κB signal pathway,exerts therapeutic effects on EAP rats.Chapter 2:Study on the anti-inflammatory mechanism of Qianlie-Biertong Suppository Objective:To investigate the effect and mechanism of Qianlie-Biertong Suppository in improving prostatitis in vitro.Methods:Qianlie-Biertong Suppository intestinal absorption solution was prepared in vitro and analyzed by UPLC-QE-Orbitrap-MS method.To establish an inflammatory model of human prostate cancer cell line PC-3 induced by lipopolysaccharide(LPS).Cell viability was measured using 3-(4,5)-dimethylthi-ahiazo(-z-yl)-3,5-di-phenytetrazoliumromide(MTT)assay;qRT-PCR was used to detect the gene expression level of inflammatory factors;Western blot was used to detect the expression level of key proteins in Toll like receptor4(TLR4)signaling pathway;using immunofluorescence technology nuclear translocation detects NF-κB p65,activator protein 1(AP-1)subunit c-Jun,and interferon regulatory factor 3(IRF3).Results:IQS mainly contains active ingredients such as berberine,chlorogenic acid,ferulic acid etc,the concentration range of ranging from 6.25 to 400 μg/ml had no significant effect on the viability of PC-3 cells.IQS(200 μg/mL and 400 μg/mL)can significantly inhibit the inflammatory factor under LPS stimulation TNF-A,IL-6,CXCL10,monocyte chemoattractant protein-1(MCP1),and COX2 genes expression.IQS(200 μg/mL and 400 μg/mL)can reduce the phosphorylation levels of protein kinase B(Akt)and NF-κB inhibitory protein α(IκB-α),IκB-α kinase α/β,p65,p38,c-Jun,TANK binding kinase 1(TBK1),and IRF3.The distribution of p65,c-Jun,and IRF3 in the cytoplasm of the model group decreased,while the distribution in the nucleus increased.IQS(200 μg/mL and 400 μg/mL)inhibits LPS induced nuclear translocation of p65,c-Jun,and IRF3.Conclusion:Qianlie-Biertong Suppository intestinal absorption solution can improve the inflammatory response of PC-3 cells induced by LPS,and its mechanism is related to the inhibition of TLR4 signaling pathway.