The Effect And Mechanism Of Ganoderma Lucidum Triterpenoid Inhibiting The Growth Of Benign Prostatic Hyperplasia

Prostate disease has become one of the major diseases threatening human health.In the United States,the incidence of benign prostatic hyperplasia increases by 10%for every 10 years of age of elderly men.However,with the aging society and the change of living habits and dietary habits in China,the incidence of benign prostatic hyperplasia is increasing year by year.And hyperplasia of the prostate can lead to a doubling of the incidence of prostate cancer.Therefore,the development of treatment methods for prostate diseases is extremely urgent.Edible and medicinal macrofungi are important biological groups with new structures and active natural small molecular compounds,and have become important resources for drug development.Ganoderma lingzhi has the functions of immune regulation,antitumor activity,protecting liver.Many studies have shown that natural products of Ganoderma lingzhi polysaccharides and triterpenes of prostate disease have good treatment recently,and the team's previous research has shown that Ganoderma lingzhi fruiting body alcohol extract with no obvious side effects,but in vivo experiments of Ganoderma triterpene compounds and prostate disease activity and mechanism of action research is less.In this paper,used Ganoderma lucidum triterpenoids by hot reflux extraction?HRE?and supercritical carbon dioxide extraction?SFE-CO₂?methods,studied the effluence on prostatic hyperplasia in mice induced by testosterone propionate.AlamarBlue?method,flow cytometry?Annxin V-FITC?/PI double staining method and HPLC method were used to evaluate anti-tumor activity of Ganoderma lucidum triterpenoids against LNCaP in vitro.Flow cytometry 2',7'-dichlorofluorescindiacetate?H2DCFDA?staining,quantitative real-time PCR?qPCR?and western blot?WB?were used to explore the mechanism of anti-tumor action.which can provide a theoretical basis for the screening model of relevant targeted active substances.The results are as follows:According to the effect of Ganoderma lucidum extract on testosterone propionate-induced prostate hyperplasia in mice,it was concluded that Ganoderma lucidum extract can reduce the content of androgen T and DHT in androgen-dependent benign prostatic hyperplasia in mice,and inhibit The expression of AR,and the inhibition of the expression of PSA and PAP closely related to the prostate,can inhibit the release of FSH and LH in the hypothalamic-pituitary-testis interstitial axis,thereby inhibiting the androgen feedback regulation.In addition,the observation of prostate morphology in experimental mice proved that Ganoderma lucidum triterpenoid extract can not only inhibit the expression of prostatic hyperplasia related hormones,but also inhibit the development of benign prostatic hyperplasia and improve prostatic hyperplasia cells.According to Ganoderma lucidum A,Ganoderma lucidum D,Ganoderma sterol triol,Ganoderma lucidum DM,Ganoderma lucidum S,Ganoderma lucidum B,six kinds of Ganoderma lucidum triterpenoids for the proliferation inhibition test of prostate cells LNCaP,the six Ganoderma lucidum triterpenoids were obtained.The compound inhibited androgen-induced prostate cell LNCaP.The inhibitory effect of ganoderic acid A was 52.3%,and the inhibitory effects of ganoderic acid D and ganodester triol were42.1%and 45.7%,respectively.In this experiment,HPLC,RT-qPCR and western experiments were used to test the mRNA and protein expression of 5-?reductase and AR by three kinds of Ganoderma lucidum triterpenoids such as Ganoderma lucidum A,Ganoderma lucidum D,Ganoderma glycerol and Ganoderma lucidum DM.The results showed that the four Ganoderma lucidum compounds could inhibit the action of 5-?reductase.Among them,300?M Ganoderma ketone triol had the best inhibitory effect,which was 61.3%,while the other three compounds had the best inhibitory effect at 50.%about.In addition,Ganoderma lucidum A,Ganoderma lucidum D and Ganoderma lucidum triterpenoids can inhibit the androgen-induced AR and AR protein expression in LNCaP cells,and the inhibitory effect on AR protein expression is concentration-dependent.Sexuality,300?M of Ganoderma lucidum D has the most significant inhibitory effect,far less than the negative control,only one tenth of it.The experimental results also confirmed that Ganoderma lucidum A,Ganoderma lucidum D and Ganoderma glycerol can induce apoptosis of LNCaP in prostate cells and inhibit LNCaP cell proliferation in a concentration-dependent manner.When the concentration was 300?M,the early apoptotic rate induced by ganoderma ketol was8.5%,the total apoptotic rate was 9.9%,the early apoptotic rate induced by ganoderic acid A was 6.3%,and the total apoptotic rate was 7.5%.The early apoptotic rate induced by acid D was 5.3,and the total apoptotic rate was 6.3%,which may be one of the important reasons for the inhibition of apoptosis of LNCaP cells by Ganoderma lucidum compounds.In addition,these three kinds of Ganoderma lucidum can also induce the release of ROS in LNCaP cells,and increase with this concentration,which may be one of the important reasons for the induction of apoptosis by three Ganoderma lucidum compounds.In this experiment,the expression of apoptotic genes and proteins was also determined in cells treated with Ganoderma lucidum A,Ganoderma lucidum D and Ganoderma glycerol.The results showed that the treatment of Ganoderma lucidum A resulted in up-regulation of mRNA and protein expression levels of caspase-3,bad and aifm1 in LNCaP cells.It is indicated that Ganoderma lucidum A promotes the expression of Bad,Caspase-3 and AIFM1 in LNCaP of prostate cells,which may be one of the important reasons for promoting apoptosis of prostate cancer cells.The ganoderma ketone triol group showed that the mRNA and protein expression levels of caspase-3,bad,aifm1 and bax were up-regulated,while the Ganoderma lucidum D group could up-regulate the expression of caspase-3 and bad genes and proteins,and inhibit bcl-2.The expression of genes and proteins may be one of the important reasons for their induction of apoptosis in prostate cell LNCaP.