| Objective To observe the change of HPG axis related hormone and testosterone synthesis enzyme in rats,testosterone metabolism related factors in track and field athletes by using the Formula of TCM Extracts of Tonifying Kidney and Benefiting Yuan,as a new Chinese medicine preparation were contrasting with Herba Epimedii extractive to compare the interventional effect,To explore the effect of this prescription on exercise-induced low serum testosterone in rat model from the secretion,synthesis and metabolism of testosterone,in addition,the intervention effects of formula of TCM Extracts were compared with single herb extracts,so as to provide a possible theoretical basis for the clinical development of new sports nutrition supplements.At the same time,the preliminary study of the main effective components has been finished.Methods The contents of panaxoside and icariin were determined by double wavelength HPLC method with gradient elution;The content of total polysaccharide in the compound were determined by UV method.40 Male 2 months old Sprague-Dawley rats were divided randomly into CS,MS,ME,MLC and MHC group.All rats except those in CS group treadmill training every day with incremental load for 6 weeks,By the Elisa detection with the content of serum CORT and T hormone;The ultrastructure of testis in rats were observed by transmission electron microscope;The mRNA expression and protein expression level of FSH,LH,GnRH in brain tissue and HMG-CoA reductase,LDL-R,SR-BI,CYP11A1,StAR and LH/CG in testicular Leydig cells were determined by real-time PCR and western-blot;The expression level of 17-KS in the testosterone metabolism of athletes were determined by UV method.Results(1)The 11 active components of panaxoside and icariin could be clearly separated by HPLC.Each capsule contains 43.57 mg of total saponins of ginseng and 5.03 mg of icariin were calculated according to the external standard method.Each capsule contains 0.268 g of the polysaccharide by UV method and by standard curve method.(2)After vigorous exercise,the ratio of serum testosterone to cortisol,serum testosterone and hemoglobin in the MS group were significantly decreased(P < 0.01),the blood testosterone value decreased by 38%compared with the CS group.Blood lactate and CK increased significantly(P <0.01),and BUN increased obviously(P<0.05),and cortisol showed no significant change(P>0.05).While T,T/C ratio and Hb increased to different degrees in the three groups.BLA,CK and BUN decreased to different degrees.The results showed that the rat model of exercise-induced low serum testosterone was successfully established.(3)Leydig cell edema,chromatin edge accumulation,cell nucleus pyknosis,endoplasmic reticulum to expand,mitochondria partially vesicles were seen under the lens in the MS group.Compared with the MS group,the changes of Leydig cells included a few mild swelling of mitochondria and no obvious change of endoplasmic reticulum under the transmission electron microscopy in the drug group.(4)The mRNA and protein expression levels of HMG-Co A,St AR and CYP11A1 in ME,MLC and MHC groups were significantly higher than the MS group(P<0.01),but the mRNA and protein expression levels of LDL-R and SR-BI were not significantly different in MS group(P > 0.05).(5)The mRNA and protein expression levels of FSH,LH,GnRH and LH/CG in MS group were significantly lower than CS group(P < 0.01).The mRNA and protein expression levels of FSH、LH、Gn RH and LH/CG in drug group were significantly higher than that in the MS group(P <0.01).(6)Comparedwith MS group,the content of 17-KS in urine at 24 h in the ME,MLC and MHC groups showed no significant change(P>0.05),but showed an upward trend.Compared with MS group,the content of 17-KS in blood in the ME,MLC and MHC groups were significantly increased successively(P<0.01),but it was still lower than before the experiment.Conclusion(1)The formula of TCM extracts of Tonifying Kidney and Benefiting Yuan,The 11 components,including ginsenoside and icariin were effectively separated,and the polysaccharide was detected.The method was feasible and simple operation,high specificity,good reproducibility,and could be used for internal quality control of this compound.(2)The model of low-blood testosterone can be successfully established by using the running table with gradually increasing load.Based on the observation of the general situation of rats,the detection of serological indexes and the observation of the morphology of testicular interstitial cells,it is proved that the formula of TCM extracts of Tonifying Kidney and Benefiting Yuan has the effect of preventing and treating exercise-induced lower blood testosterone in rats.(3)By constructing motility in the rat model with low blood testosterone,The mechanism of action of the formula of TCM extracts of Tonifying Kidney and Benefiting Yuan against exercise-induced lower blood testosterone mainly focuses on the effect on testosterone secretion and synthesis in rats.This includes that(1)Two key steps of endogenous cholesterol synthesis and testosterone synthesis in testicular Leydig cells can be used to influence blood testosterone concentration,but the molecular mechanism of cholesterol synthesis influenced by LDL-R transcription and/or SR-BI mediated reverse transport needs to be further explored.(2)It has a multi-link and multi-target promoting effect on the HPG axis and affects the concentration of blood testosterone from the perspective of affecting the secretion of testosterone. |
