| Objective: To investigate the regulation mechanism of Yishen Tonglong Decoction on EMT of benign prostatic hyperplasia through the AKR1B10-NF-k B inflammatory pathway.Methods: The experiment was designed from both cell and animal levels.Cell experiments:(1)The background expressions of AKR1B10 and NF-k B in BPH-1 cells and RWPE-1 cells were compared,and the changes in the expression levels of the two target proteins in different cells were detected by Western blot.Then the stable cell lines were selected according to the background expressions.(2)Over expression and silencing of Ak R1B10 and NF-k B genes were performed to observe the phosphorylation level of NF-k B p65 and the expression of E-cadherin protein under different cell conditions.Animal experiment: mice were randomly assigned to the normal group,model group,and the low-dose,medium-dose and high-dose groups of Yishen Tonglong Decoction.Mouse prostatic hyperplasia model was prepared by intraperitoneal injection of low-dose testosterone propionate injection.After successful modeling,normal group and model group were given normal saline intragastric administration,and each treatment group was given corresponding dose of Chinese medicine intragastric administration,for 30 consecutive days,once a day.The mice were sacrificed 24 h after the last administration,and the prostate tissues were collected for detection of related indexes.SPSS 20.0 statistical analysis software was used to process the original data,and professional conclusions were drawn according to the statistical results of each index.Results: Cell experiment: The background expression results show that there are differences in the contents of AKR1B10 and NF-k B in different cell environments,and the expression levels of the two in BPH-1 cells are significantly higher than those in RWPE-1 cells.The difference in the phosphorylation level of NF-k B p65 and the expression of E-cadherin protein in the total protein of BPH-1 + AKR1B10 overexpression cells and BPH-1 + Vector empty cells was statistically significant(P < 0.05).There were significant differences in NF-KB p65 phosphorylation level and E-cadherin protein expression between BPH-1 + AKR1B10 si RNA cells and BPH-1 + si RNA NC cells(P<0.05).The expression of E-cadherin protein in BPH-1 + Vector no-load cells and BPH-1 + Vector no-load +BAY11-7082 5UM cells was significantly different(P<0.05).Animal experiment: The mouse model of prostatic hyperplasia was successfully prepared by intraperitoneal injection of small dose of testosterone propionate injection.The expression of AKR1B10,NF-k B P65 and its downstream components(IL-8,HIF-1α,IL-1β,TNF-α)and EMT-related proteins(E-cadherin,N-cadherin)were significantly different between model group and normal group(P<0.05).Each dose of disease-enhancing decoction can reduce the protein expressions of AKR1B10,NF-k B P65,IL-8,HIF-1α,IL-1β,TNF-α,and N-cadherin,and enhance the protein expression of E-cadherin.Among them,the medium dose can improve the expression level of related factors most obviously,and the difference between the low dose and the high dose groups is also statistically significant(P<0.05).Conclusion:1.Mice prostatic hyperplasia model can be successfully prepared by intraperitoneal injection of small dose of testosterone propionate injection.2.Inflammation-induced epithelial mesenchymal transformation is a key process in the development and progression of BPH.EMT mediated by the inflammatory pathway of AKR1B10-NF-k B may be the pathogenic mechanism of BPH,in which Ak R1B10 may be the upstream link that initiates the inflammatory pathway of NF-k B in the pathogenesis of BPH induced by inflammation-induced EMT,while the inflammatory pathway of NF-k B may be the key process in the pathogenesis of BPH induced by inflammation-induced EMT.3.Yishen Tonglong Decoction may be effective in the treatment of BPH by interfering with EMT mediated by the AKR1B10-NF-k B inflammatory pathway.Low dose of Yishen Tonglong Decoction is the optimal therapeutic dose. |
PDF Full Text Request