Study On The Metabolic Pathway Of Kangquan Recipe In Vivo Based On Its Inhibition Of BPH-1 Cell Proliferation

AIM:In this study,nuclear magnetic resonance metabonomics technology and molecular biology experiment were used to study the effect of Kangquan recipe medicated serum on the proliferation of BPH-1 and the changes of metabolite spectrum of prostate,liver and blood in BPH rats,so as to explore the potential metabolic pathway of Kangquan recipe in inhibiting BPH.Methods:First of all,through cell experiment,it was verified that Kangquan recipe containing serum inhibited the proliferation of BPH-1 cells and promoted their apoptosis,so as to find out the possible mechanism.Next,we verified the therapeutic effect of Kangquan recipe on BPH in rats.Finally,we used metabonomics to study the biochemical differences and metabolite effects of biological fluid(serum)and tissue extracts(liver and prostate)in rats with BPH.Finally,cell experiment and animal experiment were combined to find the metabolic pathway of Kangquan recipe in the treatment of BPH.Results:Cell experiment:According to different groups,rats were given corresponding drugs by gavage for 30 days.Arterial blood was collected from rats in each group to prepare drug-containing serum of Kangquan recipe,the prepared drug-containing serum was filtered and cultured into BPH-1 cells.Then carry out related experiments.MTT and EDU experiments showed that Kangquan recipe medicated serum could inhibit the proliferation of human BPH-1 cells;The content of ROS in BPH-1 cells cultured with serum containing Kangquan recipe increased.Cell cycle detection experiments showed that Kangquan recipe medicated serum caused BPH-1 cell cycle arrest;The content of ROS in BPH-1 cells cultured with serum containing Kangquan recipe increased.Western blot and qRT-PCR results showed that Kangquan recipe medicated serum could affect the expression of cell cycle and apoptosis related proteins and mRNA.Animal experiment in vivo:Firstly,the model of BPH was established in SD rats,and the rats were treated with high,medium and low doses of Kangquan recipe respectively.The results showed that Kangquan recipe could reduce the wet weight of prostate tissue in BPH rats,and the pathological section of prostate tissue showed that the prostate tissue of BPH rats treated with Kangquan recipe had mild hyperplasia.Metabonomics results:After treatment,Kangquan recipe could reduce the contents of low-density lipoprotein,acetic acid,acetone,phosphocholine,glycerophosphatidyl-choline and lipids in serum of rats,and the contents of glutamic acid and lactate returned to normal levels compared with the model group.At the same time,Kangquan recipe can also reduce the contents of phosphocholine,glycogen,βglucose,lysine,serine and lactate in liver,reduce the contents of taurine,glutamine,citrate,glutathione and aspartate in prostate tissue,and increase the content of formic acid.Conclusion:This study found that Kangquan recipe can achieve the purpose of treating BPH at both animal level and cell level.Through metabonomics analysis,the mechanism of Kangquan recipe was deeply explored,which provided a new breakthrough for the treatment of BPH with traditional Chinese medicine.