Study On The Effect And Mechanism Of Xianjia Decoction On Prostatic Hyperplasia Based On IL-6/STAT3/miR-181b-1/CYLD Pathway

First of all,this study summarized the previous data,proposed that kidney deficiency and blood stasis were the pathological basis of benign prostatic hyperplasia,and explored the relationship between kidney deficiency and blood stasis and inflammation,as well as the mechanism of the treatment of benign prostatic hyperplasia by tonifying the kidney and removing blood stasis;To study the efficacy and safety of Xianjia Decoction in treating BPH of kidney deficiency and blood stasis type;Prostatic hyperplasia rats were constructed to screen specific miRNAs;To explore the therapeutic effect and mechanism of Xianjia decoction on rats with benign prostatic hyperplasia from the signal pathway of IL-6/STAT3/miR-181b-1/CYLD,and provide theoretical basis for the treatment of BPH.Theoretical research: This paper summarized the past and present studies of traditional Chinese medicine experts on BPH,and concluded that kidney deficiency and blood stasis were the basic pathogenesis of BPH,and chronic inflammatory reaction was one of the pathogenesis of BPH,while kidney deficiency and blood stasis were closely related to the occurrence of inflammatory reaction.Therefore,the treatment of BPH should be based on tonifying the kidney and removing blood stasis.Clinical study:Purpose:The patients with benign prostatic hyperplasia of kidney deficiency and blood stasis type were taken as observation to study the effect of Xianjia Decoction on patients with benign prostatic hyperplasia.Method:60 patients with benign prostatic hyperplasia(kidney deficiency and blood stasis type)were selected from the urology outpatient department of Hubei Provincial Hospital of Traditional Chinese Medicine and the outpatient department of the Affiliated Hospital of Hubei University of Traditional Chinese Medicine from September 2021 to September 2022.They were randomly divided into treatment group and control group with 25 cases each.The experimental group was treated with Xianjia Decoction,and control group was treated with finasteride for 8 weeks.The International Prostate Symptom(IPSS)score,Chinese Medicine Symptom Score,Quality of Life Score(QOL),Maximum Urinary Flow Rate(Qmax),Prostate Volume(PV),Bladder Residual Urine Volume(RUV),International Erectile Function Score(IIEF-5)and curative effect of Chinese medicine were evaluated before and after treatment,and the data were statistically analyzed.Results: In this study,50 patients with BPH of kidney deficiency and blood stasis type were selected according to the inclusion,exclusion and exclusion criteria.Compared with before treatment,the IPSS,TCM syndrome score,QOL,PV and RUV in the experimental group decreased significantly after treatment,while IIEF-5 and Qmax increased significantly(P<0.01);Compared with before treatment,the IPSS,TCM syndrome score,QOL,PV,RUV of the control group were significantly lower,Qmax was significantly highe(P<0.01),and IIEF-5 was lower,but the difference was not statistically significant(P>0.05);Compared with the control group after treatment,the IPSS,TCM symptom score,QOL,PV and RUV of the experimental group were significantly lower,and the IIEF-5 and Qmax were significantly higher(P<0.05 or P<0.01);In the experimental group,0 cases were recovered,10 cases were evidently effective,13 cases were effective,and 2 cases were effectless.The total effective rate was 92%;In the control group,0 cases were cured,5 cases were evidently effective,13 cases were effective,and 7 cases were ineffective.The total effective rate was 72%;The treatment effect of the experimental group is better than the control group(P<0.05).Conclusion:1.Xianjia decoction is better than finasteride in treating BPH patients with kidney deficiency and blood stasis;2.Both Xianjia decoction and finasteride can improve the clinical symptoms of patients and reduce the volume of prostate,but Xianjia decoction is better than finasteride;3.Xianjia Decoction can also improve the erectile function of BPH patients and make them more acceptable;4.Xianjia Decoction has no adverse reactions in clinical treatment of BPH,which is worthy of clinical application.Experimental study:Purpose:We used rats with benign prostatic hyperplasia to study the mechanism of Xianjia Decoction in treating benign prostatic hyperplasia through inflammation mediated by mast cells,IL-6/STAT3/miRNA-181b-1/CYLD signal pathway.Methods: 60 SPF male rats were randomly divided into 6 groups(control group,model group,high/medium/low dose group of Xianjia decoction,and western medicine group).The rats in the control group were subcutaneously injected with normal saline,and the rats in the other groups were subcutaneously injected with testosterone propionate(5mg/kg);Once a day,after 30 days of continuous modeling,confirm whether the modeling is successful;If the BPH model is successfully constructed,continue to give drug treatment: The intragastric dose was 2.718g/100 g in the high dose group,1.359g/100 g in the middle dose group and 0.6795g/100 g in the low dose group;The western medicine group was gavaged with finasteride 0.45mg/kg daily.Once a day,the administration period was 30 days,and then the rats were killed.Observe the changes of vitality,hair gloss and padding humidity of rats;Weigh the rats’ body weight and prostate weight,calculate the prostate index;HE staining was used to detect the pathological changes of prostate tissue in each group;The number of mast cells in prostate tissue was detected by toluidine blue staining;Immunoblotting was used to detect the protein expression of STAT3,p-STAT3,JAK2,p-JAK2;The expression of IL-6 and IL-10 in prostate tissue was detected by ELISA kit;The differential expression of miRNA in prostate tissue between the control group and the model group was detected and analyzed by miRNA sequencing;RT-PCR was used to detect the expression of miR-181b-1-3p and CYLD m RNA in prostate tissue of each group.Results:The rats in the control group were active,their hair was bright,and the padding was clean and dry;The rats in the model group and the low-dose group of Xianjia Decoction were easily agitated,the hair gloss was average,and a few were injured in fights,and their living environment is more humid;Compared with the model group,the rats in the high/medium dose groups and the western medicine group were in good mental state,and the hair gloss and pad humidity were improved compared with the model group,without obvious fighting.There was no significant difference in body weight of rats in each group(P>0.05);Compared with the blank group,the volume,wet weight and index of prostate in the model group were significantly higher(P<0.01);Compared with the model group,the volume,wet weight and index of the prostate in the high dose group,the middle dose group and the western medicine group of Xianjia Decoction decreased significantly(P<0.05 or P<0.01),while there was no significant difference in the low-dose group(P>0.05);Compared with the western medicine group,the volume,wet weight and index of prostate in the high dose group of Xianjia Decoction decreased significantly(P<0.05 or P<0.01).The HE staining results of prostate tissue in each group showed that the glands in the blank group were full and regular,the glandular epithelium was monolayer columnar cells,and the matrix tissue was loose;In the model group,the glands were irregular,the glandular epithelial cells proliferated,some of them were monolayer high columnar,forming mucosal folds,like papillary processes,fibrous tissue hyperplasia in the matrix,and inflammatory cell infiltration;In the high-dose group of Xianjia Decoction,the glands were regular,no obvious glandular epithelial hyperplasia was found,a few papillary processes,a small amount of fibrous tissue hyperplasia in the matrix,and a small amount of inflammatory cell infiltration;In the middle dose group of Xianjia Decoction,the glands are regular,some of them have papillary protrusions,fibrous tissue hyperplasia in the matrix,and some of them have inflammatory cell infiltration;In the low-dose group of Xianjia Decoction,the glands were irregular,with more papillary processes,hyperplasia of matrix fibrous tissue,and inflammatory infiltration;In the western medicine group,the glands are regular,with papillary processes,and some of them have hyperplasia of stromal fibrous tissue and inflammatory infiltration.There were more mast cells in the prostate tissue of the model group than that of the blank group,the difference was statistically significant(P<0.01);Compared with the model group,the number of mast cells in the high and middle dose groups of Xianjia Decoction and the western medicine group decreased significantly(P<0.05 or P<0.01);Compared with the western medicine group,the number of mast cells in the high dose group of Xianjia Decoction was significantly reduced(P<0.05).Compared with the blank group,IL-6 in the prostate tissue of the model group was significantly higher and IL-10 was significantly lower(P<0.01);Compared with the model group,the levels of IL-6 and IL-10 in the high and middle dose groups of Xianjia Decoction and western medicine groups were significantly lower and higher(P<0.01);Compared with the western medicine group,the IL-6 in the high dose group of Xianjia Decoction was significantly lower and the IL-10 was significantly higher(P<0.01).Compared with the blank group,p-STAT3 and p-JAK2 in the prostate tissue of the model group were significantly higher(P<0.01);Compared with the model group,the levels of p-STAT3 and p-JAK2 in the high and middle dose groups of Xianjia Decoction and the western medicine group were significantly lower(P<0.05 or P<0.01);Compared with the western medicine group,p-STAT3 and p-JAK2 in the high dose group of Xianjia Decoction were significantly lower(P<0.05).According to the statistics of sample sequencing data volume,base quality score,s RNA length and type distribution,and miRNA base preference identification,the quality of miRNA sequencing data in this study is well controlled,and the obtained s RNA is mainly miRNA.Through preliminary prediction and filtration of miRNA target genes,13571 target genes were found;Through the analysis of the differential miRNA in the prostate tissue of the two groups,it was found that the miRNA in the model group was up-regulated by 18 and down-regulated by 17 compared with the blank group.GO enrichment analysis: A total of 418 GO enrichment results were involved,including 164 biological processes,199 molecular functions,and 55 cell components,of which 26 were statistically significant differences(including 8BP,13 MF,and 5 CC)(Padj<0.05).The top 20 were selected for mapping and analysis.Among the biological processes involved,viral process was the most enriched,followed by DNA fusion,microtubule process,and so on,Among the molecular functions involved,metalloproteinase activity is the most enriched,followed by microtubule binding and metalloendopeptidase activity,and the cell components involved are the plasma membrane,followed by microtubule and kinetin complex.KEGG enrichment analysis: A total of 246 KEGG enrichment pathways were involved,of which 22 were statistically significant(Padj<0.05).The first 20 pathways were selected for mapping and analysis,which involved 10 biological systems,4 metabolism,6 information processes,and 1 cell process.The main pathway involved is PI3K/AKT signal pathway,followed by MAPK signal pathway,calcium signal pathway,Hippo signal pathway,etc.Compared with the blank group,the expression of miR-181b-1-3p in the prostate tissue of the model group increased and the expression of CYLD m RNA decreased,and the difference was statistically significant(P<0.01);Compared with the model group,the expression of miR-181b-1-3p and CYLD m RNA in the high and middle dose groups of Xianjia Decoction and the western medicine group decreased significantly(P<0.05 or P<0.01),while the expression of miR-181b-1-3p and CYLD m RNA in the low dose group of Xianjia decoction decreased significantly(P>0.05);Compared with the western medicine group,the expression of miR-181b-1-3p and CYLD m RNA in the high dose group of Xianjia Decoction decreased significantly(P<0.05 or P<0.01).Conclusion: 1.Xianjia decoction can smooth the mood of rats and improve the urinary tract symptoms of rats;Xianjia decoction can reduce the prostate volume of BPH rats,reduce the wet weight and index of BPH rats,and the high-dose group can play a better therapeutic effect than the western medicine group;Xianjia Decoction can improve the histopathological condition of BPH,and high dose has the best effect.2 Xianjia decoction can improve BPH by inhibiting mast cells and IL-6/STAT3 signal pathway,and the effect is more obvious than that of western medicine.3.Multiple miRNAs are related to the pathogenesis of BPH,and the target genes regulated by them participate in multiple biological processes,molecular functions,cell components and signal pathways.3.Xianjia Decoction can reduce the inflammatory reaction of prostate tissue,inhibit cell proliferation,promote apoptosis,and improve BPH by regulating the positive feedback loop of IL-6/STAT3/miRNA-181b-1/CYLD,and the high dose of Xianjia Decoction has better curative effect than western medicine.