Studies On The Bioactive Fraction Of Prinsepia Utilis Royle Leaves Extracs On Inhibition Of Benign Prostatic Hyperplasia

Prinsepia utilis Royle belongs to prinsepia genus?rosaceae?,mainly distributed in southwest China such as Yunnan and Guizhou.Pharmacological studies have shown that the stems and leaves of P.utilis had the functions of regulating blood lipids,anti-oxidation,anti-osteoporosis,anti-tumor,et al.P.utilis leaves extracts?QCJ?was also confirmed to improve benign prostatic hyperplasia?BPH?previously in our laboratory.To clarify the active components and mechanism of QCJ on anti-BPH,the BPH model was built on the castrated rats by subcutaneous injection of testosterone propionate.The anti-BPH activity of QCJ and its three fractions?Fr.A,Fr.B,Fr.C?were evaluated.The prostatic index,prostate tissue morphology,serum prostatic acid phosphatase?PACP?and dihydrotestosterone?DHT?levels were used as indicators to screen the active fraction of BPH in rats.To investigate the anti-BPH mechanism of QCJ and its effective fraction,the expression of vascular endothelial growth factor?VEGF?in the prostate tissue of rats was tested,and serum levels of testosterone?T?,estradiol?E₂?,interleukin-6?IL-6?,tumor necrosis factor-??TNF-??and total superoxide dismutase?SOD?,glutathione peroxidase?GSH-Px?,catalase?CAT?,malondialdehyde?MDA?in prostate homogenate were measured.Furthermore,the chemical compositions of effective fraction were characterized by UPLC-Q-TOF-MS/MS and partially confirmed by further isolation and identification with spectroscopic analysis.The results showed that the anti-BPH activity of Fr.B was the closest to QCJ in all fractions,which significantly decreased pathological changes in the prostate tissue,reduced the prostatic index,serum PACP and DHT?P<0.01?.Both QCJ and Fr.B significantly reduced the expression of VEGF?P<0.01?and MDA?P<0.01?levels in prostate tissue,as well as DHT?P<0.01?content,DHT/E₂?P<0.01?ratio and TNF-??P<0.01?content in serum of BPH rats.Besides,QCJ and Fr.B increased SOD?P<0.01?,GSH-Px?P<0.05?and CAT?P<0.01?levels in prostate tissue and T?P<0.01?content in serum of BPH rats.In addition,ten flavonoid glycosides were assigned from Fr.B by UPLC-Q-TOF-MS/MS,which accounted for 54.96%of all substance of Fr.B.Three flavonoid glycosides were further isolated from Fr.B and identified by spectroscopic analysis as Kaempferol-3-O-[?-L-rhamnopyranosyl-?1?6?-[?-L-rhamnopyranosyl-?1?2?]-?-D-glucopyranosyl]-7-O-?-D-glucopyranoside?1,peak 2?,Kaempferol 3-O-[?-L-rhamnopyranosyl-?1?6?-[?-L-rhamnopyranosyl-?1?2?]]-?-D-glucopyranoside?2,peak 6?and Kaempferol 3-O-[?-L-rhamnopyranosyl-?1?6?]-?-D-glucopyranoside?3,peak 9?.Compound 1 has not been reported before,and compound 2 is isolated from P.utilis for the first time.This study demonstrated that Fr.B was an effective fraction of QCJ on the inhibition of BPH.The mechanism of QCJ and Fr.B may be relevant to downregualtion of the ratio of DHT to T and upregulation of T level in BPH rats serum,inhibition of the VEGF expression of prostate,and increasing the anti-inflammatory and anti-oxidant levels,the flavonoids might be the pharmacological ingredients of P.utilis leaves.This study provided scientific basis and supported for clinical application of P.utilis for BPH treatment.