| Chronic prostatitis is a common clinical disease,which seriously affects men's physical and mental health.In recent years,with the change of lifestyle,the incidence rate has increased year by year.? Prostatitis accounted for more than 90%of chronic prostatitis,mainly manifested as pelvic pain,discomfort and abnormal urination.The clinical manifestations were persistent,refractory and site variability.Antibiotics and ?-blockers are the representatives of Western medicine treatment.Although they can alleviate some symptoms of patients to a certain extent,the treatment effect is non-ideal.Traditional Chinese medicine has certain advantages because of its multi-channel and multi-target therapeutic effect.Professor Guo Jun,based on the theory of "sore-ulcer-resolving therapy",formulated Guihuang prescription for ?prostatitis with damp heat and blood stasis syndrome.Guihuang prescription has achieved certain curative effect in the early clinical practice,so it is necessary to further evaluate the effectiveness and safety of Guihuang formula through clinical observation.The possible mechanism of Guihuang prescription in the treatment of ? prostatitis was confirmed by animal experiments.ObjectiveTo research the efficacy and safety of Guihuang prescription in the treatment of ?prostatitis with damp heat and blood stasis syndrome,and to explore the possible mechanism of Guihuang prescription in the treatment of ? prostatitis through network pharmacology and animal experiments,so as to provide evidence for the clinical application of Guihuang prescription in the treatment of ? prostatitis.Method1.Clinical research66 patients diagnosed ? prostatitis with damp heat and blood stasis syndrome in the Andrology Department,Xiyuan Hospital of China Academy of Chinese Medical Sciences were randomly divided into experimental group(Guihuang prescription group)and control group(tamsulosin hydrochloride sustained-release capsule group).The patients were treated for 6 weeks and followed up for 2 weeks.NIH-CPSI score and TCM symptom score were used as the evaluation of curative effect.At the same time,the safety was evaluated.2.Prediction of network pharmacology mechanismThrough network pharmacology and molecular docking methods,the main active components,possible targets and enrichment pathways of Guihuang prescription in the treatment of ? prostatitis were predicted,and the possible mechanism of Guihuang prescription in the treatment of ? prostatitis was predicted,so as to provide ideas and theoretical basis for further experimental research.3.Experimental studyThe rat model of autoimmune ? prostatitis was established by in situ injection of the mixture of prostate protein extract and complete Freund's adjuvant of SD rats.SD rats were divided into blank group,sham operation group,model group,Guihuang low-dose group,Guihuang medium dose group and Guihuang high-dose group,with 8 rats in each group.The blank group and the sham operation group were given normal saline by oral gavage for 30 days.The low,medium and high dose groups were given Guihuang prescription by gavage at 4.9 g/kg·d,9.8 g/kg·d and 19.6 g/kg·d,respectively.After 30 days,the samples were taken to calculate the organ indexes of prostate,spleen,and thymus.The pathological changes of prostate tissue were observed by HE staining and scored.The infiltration of CD3+and CD45+cells in the prostatic stroma was observed by immunohistochemistry and detected by ELISA The levels of serum IL-6,COX-2,TNF-? and MCP-1 pro-inflammatory cytokines were detected.The expression of PI3K,Akt and p65 NF-?B mRNA in prostate tissue was detected by RT-PCR.The expression of PI3K?p-PI3K?AKT?p-AKT?P65 NF-?B?p-P65 NF-?B protein in prostate tissue was detected by Western blot.Result1.Clinical findings(1)66 cases(33 experimental,33 control)were enrolled,5 shedding cases(2 experimental,3 control)were excluded,3 excluded cases(1 experimental,2 control)and 58 cases(31 experimental,27 control)were finally eligible for the protocol set(PPS).(2)The NIH-CPSI score decreased from 28.62±8.61 to 10.58±4.03 at the end of treatment in the experimental group and from 26.74±7.72 to 13.44±5.92 at the end of treatment in the control group.The NIH-CPSI pain scores,urination scores,quality of life score and total scores(P<0.05)and the TCM symptom scores(P<0.05)were reduced in the experimental group compared with the control group after the end of treatment and before treatment.(3)The experimental group was superior to the control group in reducing NIH-CPSI pain scores,quality of life scores,and total scores(P<0.05),in reducing TCM symptom scores(P<0.05),and in decreasing NIH-CPSI urination scores(P>0.05)during treatment.(4)3 patients in the experimental group experienced gastric discomfort while taking the medicine,and the discomfort disappeared after taking the medicine after the meal;2 patients in the control group experienced transient hypotension after taking the medicine,and the discomfort disappeared after taking the medicine at bedtime,and the remaining patients showed no obvious adverse reactions during treatment.2.Network pharmacology mechanism prediction results(1)143 major active ingredients and 278 action targets of Guihuang prescription were screened,111 possible action targets of Guihuang prescription in the treatment of ? prostatitis were obtained after mapping with ? prostatitis targets.(2)The PPI interaction network analysis revealed that the genes with the most frequent protein interactions were mainly involved in apoptosis,oxidative stress,proliferation and differentiation,which was consistent with the current mechanistic findings in ? prostatitis.(3)The results of GO analysis revealed that treatment of ? prostatitis by Guihuang prescription may occur in vesicle lumen,membrane rafts,nuclear chromatin,and other sites,and acts by regulating biological pathways such as response to bacteria derived molecules,oxidative stress,apoptosis,response to lipopolysaccharide and steroid hormones,in combination with phosphatases,protein phosphatases,and so on.The results of KEGG pathway enrichment analysis suggested that Guihuang recipe may involve several signaling pathways,including PI3K-Akt,NF-?B MAPK,mTOR,IL-17,which reflect the therapeutic characteristics of multiple pathways in TCM.(4)The compatibility between drugs in the Guihuang prescription exhibited good principal compatibility as showen in the 'drug-active ingredient target-pathway' network diagram.The molecular docking results suggested that the main active ingredients had strong binding ability with the target proteins.(5)Combined with the network pharmacology results and literature studies,we predicted that Guihuang prescription might work through the PI3K/Akt/NF-?B pathway to treat ?prostatitis.3.Experimental findings(1)The rat model of ? prostatitis can be successfully reproduced by the in situ injection method of rat prostate protein extract mixture with complete Freund's adjuvant.(2)Guihuang prescription significantly reduced the prostate index and thymus index,ameliorated histopathological changes in the prostate,and alleviated CD3+ and CD45+cell infiltration in the prostatic stroma in a model of ? prostatitis.(3)Guihuang prescription inhibited the expression of serum levels of pro-inflammatory cytokines including IL-6,COX-2,TNF-? and MCP-1 in the rat model of ? prostatitis.(4)Guihuang prescription inhibited PI3K,Akt and p65 NF-?B mRNA expression and inhibited PI3K,Akt and p65 NF-?B protein phosphorylation in the rat model of ? prostatitis.Conclusion1.Clinical study showed that the experimental group could reduce the pain score,urination score,quality of life score and total score of NIH-CPSI(P<0.05)and reduce the symptom score of TCM in patients diagnosed ? prostatitis with damp heat and blood stasis syndrome.Moreover,it was superior(P<0.05)in reducing the total NIH-CPSI scores,pain scores,quality of life scores and TCM symptom scores,while no significant statistical difference was observed in the voiding scores of NIH-CPSI.And no serious adverse reactions,which deserve to be promoted in applications.2.Network pharmacology showed that Guihuang prescription might treat ? prostatitis through several signaling pathways,including PI3K/Akt,NF-?B,MAPK,mTOR,and IL-17,through processes involving apoptosis,oxidative stress,proliferation,and differentiation.3.Animal studies showed that Guihuang prescription significantly reduced prostate index and thymus index,ameliorated histopathological changes in the prostate,attenuated CD3+and CD45+cell infiltration in the prostate stroma,and inhibited serum IL-6,COX-2,TNF-?,and MCP-1 proinflammatory cytokine levels in the model of ? prostatitis.Furthermore,Guihuang prescription inhibited PI3K/AKT/NF-?B pathway activation in the model of ? prostatitis. |
PDF Full Text Request