| Background and aim:Benign prostatic hyperplasia(BPH)is one of the most common disease conditions in elderly men,affecting～50%of men in their 50s and>80%in their 80s.Wen Pi Tang from "Beiji Qianjin Yao Fang",is a classical prescription for yang deficiency of spleen and kidney.Danggui Beimu Kushen Wan from "Synopsis of Prescriptions of the Golden Chamber",is an important prescription for the treatment of turbid sperm and white turbidity with syndrome of dampness-heat The Wen Pi Tang&Danggui Beimu Kushen Wan(W&D)is a combination treatment of symptoms and root causes for BPH.This study was intended to clarify whether the combination of Wen Pi Tang and Danggui Beimu Kushen Wan could maintain prostate homeostasis by regulating paracrine signaling based on down-regulating Rab27B,and thus inhibit benign prostatic hyperplasia.Methods:BPH animal models were induced by testosterone propionate injected into the castrated rats,then the effects of W&D on benign prostatic hyperplasia were observed in vivo.Immunohistochemistry,immunofluorescence and western blotting were taken to detect protein expressions.Scanning electron microscopy and HE staining were used to observe the morphological changes of prostate tissues in rats.Gene chips from GEO database were used to analyze BPH related genes and signaling pathways.The stable cell lines of Rab27B knockdown and overexpression were constructed,and the conditioned medium was collected to culture prostatic stromal cell to evaluate the interaction between prostatic epithelial cells and stromal cells.Secretion levels of growth factors,IGF-1,EGF and TGF-β,were detected by ELISA.Expression of growth factor receptors on plasma membrane and the activity of pathways,mTORC1,AKT and MAPK signaling,were detected by western blotting and immunofluorescence confocal.Results:1.The Wen Pi Tang&Danggui Beimu Kushen Wan can inhibit the development of benign prostatic hyperplasia.Compared with the model group,W&D significantly reduced prostate size,prostate wet weight and prostate index.HE staining showed that the epithelial cells in the model group were densely arranged and the papillary hyperplasia in the gland cavity was prominent.After the treatment of W&D,the arrangement of the glandular epithelium tended to be normal.Scanning electron microscope results showed that after being treated with W&D.the prostate gland shrank,the space between the glands increased,the smooth muscle atrophied,and then the prostate hyperplasia was significantly inhibited.2.Rab27B is a key gene in the development of benign prostatic hyperplasia.DNA microarray analysis showed that the high expression of Rab27B in BPH was about 9 times that in normal tissues.Immunohistochemical tests of clinical specimens from BPH patients showed that Rab27B was highly expressed in BPH epithelial tissues compared with adjacent normal tissues(P=0.0001).3.Rab27B regulates paracrine signaling of prostate cells.After overexpression of Rab27B in normal prostate epithelial cells,ELISA results showed the secretion levels of IGF-1,EGF and TGF-β increased.Western blotting results showed that membrane presentation of growth factor receptors,IGF-1Rβ,EGFR,TGF-βRⅡ,were enhanced,and protein phosphorylation levels of paracrine signaling pathways,AKT,mTORC1,MAPK signaling,were also increased.4.W&D can down-regulate Rab27B to maintain prostatic homeostasis.Compared with the model group,Rab27B in the BPH tissues of rats was significantly down-regulated after being treated with W&D,and the paracrine signal disorder was significantly relieved.The expression of growth factors,IGF-1,EGF and TGF-β,were decreased,and the activity of AKT,MAPK pathways were also decreased.Conclusion:The Wen Pi Tang&Danggui Beimu Kushen Wan can regulate paracrine signaling to maintain prostate homeostasis and thereby inhibit the development of benign prostatic hyperplasia by down-regulating Rab27B. |
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