Ntu 1 Echinacoside nce phev kom muaj nuj nqis hauv nas Los ntawm Targeting Hypothalamic Androgen Receptor

Zhihui Jiang 1,2, Bo Zhou 2, Xinping Li2, Gordon M. Kirby3 & Xiaoying Zhang 1,2

Txiv neej infertility yog ib qho teeb meem loj rau kev noj qab haus huv nrog kwv yees li ntawm 4.2 feem pua ​​​​ntawm cov txiv neej infertility thoob ntiaj teb. Peb txoj hauj lwm thaum ntxov tau pom tiasCistanche extracttiv thaiv cov phev puas hauv cov nas thiab qhov ntawdechinacoside cov tshuaj(ECH) yog ib qho ntawm cov khoom tseem ceeb. Ntawm no peb qhia txog lub luag haujlwm tseem ceeb rau ECH, ib yam khoom ntuj uas thim rov qab lossis tiv thaiv oligoasthen aspermia hauv nas. ECH tau soj ntsuam los ntawm HPLC, qhov ntau thiab qhov zoo ntawm cov phev raug soj ntsuam thiab cov tshuaj hormones raug txiav txim los ntawm xov tooj cua-immunosorbent assay. ECH txo cov theem ntawm androgen receptor (AR) thiab cov noob tseem ceeb steroidogenic cuam tshuam raws li kev txiav txim siab los ntawm Western blot thiab qPCR tsom xam. Kev sib cuam tshuam ntawm ECH thiab AR tau soj ntsuam los ntawm kev tsis ncaj qha ELISA thiab molecular docking. Cov txiaj ntsig tau pom tias ECH ua ke nrog hypothalamic AR hauv hnab ris ntawm Met-894 thiab Val-713 inhibits kev hloov ntawm AR los ntawm cytoplasm mus rau nuclei hauv hypothalamus. Txawm hais tias qhov kev tawm tswv yim tsis zoo ntawm kev tswj hwm poj niam txiv neej tau cuam tshuam, cov lus pom zoo tau txhawb nqa kom muaj kev tso tawm ntawm luteinizing hormone thiab testosterone tom qab txhim kho cov phev kom muaj nuj nqis. Ua ke, cov ntaub ntawv no qhia tau hais tias ECH blocks AR kev ua haujlwm hauv hypothalamus kom nce cov phev thiab tiv thaiv oligoasthenospermia hauv nas.

Kev tsim cov testosterone hauv cov qe ntshav yog tswj hwm los ntawm hypothalamic-pituitary-gonadal axis (HPG) los ntawm kev tsim cov lus qhia homeostatic loop1. Gonadotrophin-tso tshuaj hormone (GnRH), secreted los ntawm hypothalamus, tuaj yeem txhawb kev tso tawm ntawm luteinizing hormone (LH) los ntawm lub caj pas pituitary, uas ntxiv dag zog rautestosteronentau lawm nyob rau hauv testicular Leydig hlwb. Testosterone yog biosynthesized los ntawm ib tug series ntawm steroidogenic enzymes. Raws li ib qho ntawm txoj hauv kev loj, steroidogenic acute regulatory (StAR) protein tuaj yeem thauj cov roj cholesterol los ntawm cov khoom siv hauv lub cev mus rau hauv mitochondria2 qhov twg nws raug rau cov roj cholesterol sab-chain cleavage enzyme (CYP11A1), 3 -hydroxysteroid dehydrogenase (HSD3), { {6}}hydroxylase (CYP17A1), thiab 17 -hydroxysteroid dehydrogenase (HSD 17) uas catalyze hloov cov roj cholesterol mus rau testosterone3,4. Testosterone ces tsis zoo pub rov qab mus rau HPG lub ntsiab lus kom txo qis LH secretion ntxiv nyob rau hauv koob tshuaj. Cov nyhuv ntawm testosterone ntawm HPG axis tawm tswv yim voj tshwm sim los ntawm kev khi rau androgen receptor (AR), pom nyob rau hauv ob qho tib si hypothalamus thiab pituitary caj pas 5. Hauv cov nas, ablation ntawm AR thiab tsawg kawg testosterone ntau lawm ua rau qib ntawm LH thiab follicle-stimulating hormone (FSH) nce 6, qhia tias AR koom nrog kev tswj hwm ntawm qhov tsis zoo pub lub voj voog. Classical genomic mechanism ntawmtestosteroneKev taw qhia tshwm sim thaum testosterone diffuses rau hauv lub cell thiab khi rau AR. Qhov ligand-receptor complex ces translocates mus rau lub nucleus qhov twg nws khi rau androgen teb cov ntsiab lus (AREs) nyob rau hauv cov kev tswj cheeb tsam ntawm testosterone- teb cov noob los hloov lawv translocation. Testosterone kuj induces txoj kev uas tsis yog classical ntawm steroid hormone kev txiav txim, yam ntxwv los ntawm cov xwm txheej ceev uas ua rau kom lub cytosolic signaling cascades ib txwm tshwm sim los ntawm kev loj hlob yam 7,8. Classical thiab non-classical Testosterone txoj kev ob qho tib si pab txhawb nqa spermatogenesis thiab fertility. Txawm li cas los xij, kev ua haujlwm ntawm AR yog qhov tseem ceeb dua hauv txoj hauv kev classical li testosterone ua kom cov phev zoo. Echinacoside (ECH) yog ib qho ntawm cov khoom siv bioactive tau los ntawm cov nroj tsuag tshuaj ntawm Echinacea9 thiabCistanche10. Nrog ib tug dav spectrum ntawm pharmacological kev ua ub no, extracts ntawm lub Echinacea yog 1 Research Center ntawm Modern Biotechnology, Tsev Kawm Ntawv ntawm Biotechnology thiab Food Engineering, Anyang Institute of Technology, Anyang, Henan, 455000, Suav teb. 2 College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi, 712100, Suav. 3 Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Ontario, N1G 2W1, Canada. Kev xa ntawv thiab kev thov rau cov ntaub ntawv yuav tsum tau xa mus rau XZ

Cistancheechinacoside cov tshuajtuaj yeem txhim kho lub raum ua haujlwm thiab txhim khotestosterone

Yog xav paub ntxiv thov hu rau:Joanna.jia@wecistanche.com

ib qho ntawm cov tshuaj ntsuab nrov tshaj plaws hauv Tebchaws Europe thiab Asmeskas feem ntau yog vim lawv cov tshuaj antioxidant11 thiab lawv lub peev xwm los tiv thaiv tus mob khaub thuas12. Txaus siab heev,CistancheExtracts thiab ECH tau ib txwm siv los ua tus neeg sawv cev tonic los kho cov kev ua me nyuam thiab txhawb cov txiv neej kev sib deev hauv cov tshuaj suav tshuaj 10. Qee cov khoom lag luam OTC ntawmCistanche extractiontau tsim los ua cov khoom noj khoom haus thiab tau txais txiaj ntsig hauv kev lag luam noj qab haus huv ntawm Tuam Tshoj thiab qee lub tebchaws Esxias (China Food and Drug Administration) 13. Txawm li cas los xij, cov txheej txheem hauv qab ntawm ECH kev txiav txim tseem tsis meej. Txiv neej infertility yog ib qho teeb meem loj rau kev noj qab haus huv nrog kwv yees li ntawm 4.2 feem pua ​​​​ntawm cov txiv neej infertility thoob ntiaj teb14. Kev kuaj mob ntawm tus txiv neej infertility tam sim no yog raws li kev tshawb fawb ntawm cov phev zoo nrog rau kev soj ntsuam ntawm cov kab mob hauv lub cev xws li phev concentration, motility, thiab morphology15. Te estrogen-mimic Bisphenol A (BPA) yog cov kab mob kis thoob plaws ib puag ncig uas tau kawm txog nws qhov cuam tshuam rau txiv neej fertility hauv ntau hom tsiaj thiab tib neeg16. BPA cuam tshuam cov hypothalamic-pituitary-gonadal axis, inhibits cov kab lus ntawm testicular steroidogenic enzymes thiab synthesis ntawm testosterone nyob rau hauv cov txiv neej pups17, ua rau lub xeev ntawm hypogonadotropic hypogonadism18. Hauv txoj kev tshawb no, peb tshawb xyuas qhov cuam tshuam ntawm ECH ntawm cov phev zoo thiab qib tshuaj hormones. Tsis tas li ntawd, BPA tau raug xaiv los ua tus neeg sawv cev ntawm cov phev raug mob los kawm ntxiv txog kev tiv thaiv ntawm ECH tiv thaiv cov phev tsis zoo.

Cistanche deserticola muaj ntau yam teebmeem, nyem qhov no kom paub ntau ntxiv

Cov txiaj ntsig

ECH txhim kho cov phev kom muaj nuj nqis.

Te phev naj npawb hauv cauda epididymis, phev viability, thiab phev motility yog nthuav nyob rau hauv Table 1. Kev kho mob nrog 80mg/kg ntawm ECH thiab 15mg/kg ntawm testosterone propionate (TP) ho nce cov phev epididymal suav. Txawm li cas los xij, tsis muaj qhov sib txawv ntawm cov phev viability thiab phev motility.

ECH nce qib testosterone thiab LH hauv cov ntshav, encephalon ntxiv rau pituitary, thiab testis.

ECH kev kho mob tshwj xeeb nce qib LH hauv cov ntshav, encephalon ntxiv rau pituitary, thiab testis (Fig. 1A). Hauv pawg ECH(H), LH theem tau nce ntxiv los ntawm 1.1-fold hauv cov ntshav, 1.4-fold hauv encephalon ntxiv rau pituitary, thiab 1.2-fold hauv testis. Testosterone tau nce ntxiv los ntawm 1.5 thiab 1. Tom qab TP kev kho mob, qib testosterone tau nce ntxiv los ntawm 1.6, 1.8, thiab 1.

Cistanche echinacoside tuaj yeem txhim kho lub raum ua haujlwm thiab txhim kho testosterone

ECH nce steroidogenic enzyme gene qhia.

ECHnce qhov kev nthuav qhia ntawm cov steroidogenic enzymes tseem ceeb hauv cov qe ntshav (Fig. 1B). Cov qib mRNA ntawm CYP11A1, CYP17A1, HSD3 1/2, thiab HSD17 hauv pawg ECH(H) tau nce ntau dua 3-fold, thaum theem ntawm Star mRNA tsis pom qhov sib txawv tseem ceeb nrog txhua tus.ECHtreatments (p>0.05). TP tsis tau hloov pauv qhov kev qhia ntawm CYP11A1, CYP17A1, HSD3 1/2, thiab HSD17 . Qhov no tau tshwm sim nyob rau hauv daim ntawv qhia kev ntsuas kub ntawm ECH ntawm steroidogenic enzyme qhia qhia tiasECHnce mRNA theem ntawm CYP11A1, CYP17A1, HSD3 1/2, thiab HSD17, thiab tshwj xeeb tshaj yog HSD3 1/2 (Fig. 1C).

ECH tuaj yeem hla cov ntshav-hlwb barrier thiab tsawg dua ntawm cov ntshav-testis barrier.

Txhawm rau soj ntsuam cov lus tawm tswv yim androgen ntawm HPG-axis, lub peev xwm ntawmECHnkag mus rau hauv lub hypothalamus raug soj ntsuam nyob rau hauv ib tug series ntawm PK kev tshawb fawb.ECHCov concentrations tau kuaj pom nyob rau hauv hypothalamus tom qab 12h ib qho kev tswj qhov ncauj ntawm 30mg / kg ntawm ECH, thiab ECH qhia pom tias hypothalamus penetrance nrog qhov nruab nrab hypothalamus / plasma ratio ntawm 33.92 feem pua ​​(Fig. 2A). Te T1/2 qhov tseem ceeb yog 2.61±{13}}.42 h thiab 1.88±{16}}.22 h hauv cov ntshav plasma thiab hypothalamus, ntsig txog (Fig. 2B). Txawm li cas los xij, kev nkag tau tsawg heev ntawm ECH rau hauv cov qog ntshav tau kuaj pom siv HPLC. Te concentrations ntawm ECH hauv testis yog 0.

ECH txo cov hypothalamic AR translocation rau lub nucleus.

Mus kuaj cov nyhuv ntawmECHntawm AR translocation, kev qhia ntawm AR nyob rau hauv cytoplasm thiab nucleus ntawm testis thiab hypothalamus tau txiav txim siab siv western blot tsom xam. Raws li qhia hauv daim duab 3A, ECH down-regulated AR protein nyob rau hauv hypothalamic nuclei los ntawm tsib-fold piv rau pawg tswj. Kev kho mob nrog enzalutamide, AR inhibitor, txo qis AR protein nyob rau hauv hypothalamic nuclei los ntawm 4.8-fold. Qhov tseem ceeb, cytoplasmic AR tau siab dua nrog ECH kev kho mob thiab kev kho enzalutamide dua li kev tswj hwm, qhia tiasECHthaiv AR thauj los ntawm cytoplasm mus rau lub nucleus hauv hypothalamus. Raws li qhov xav tau, ECH tsis cuam tshuam AR kev thauj mus los ntawm cytoplasm mus rau lub nucleus hauv testis (Fig. 3B).

ECH tau nce cov kev qhia ntawm HPG-axis ntsig txog cov noob.

ECHKev nce ntxiv ntawm LH, LHR, GnRH 1, thiab Gnrhr hauv cov pas dej sib xyaw ntawm encephalon ntxiv rau pituitary, txawm li cas los xij, tsis muaj qhov sib txawv tseem ceeb hauv GnRH 1 qib tom qab kho TP. Kev tsom xam ntawm cov cua sov qhia tau hais tias cov kab lus ntawm GnRH 1 thiab LH yog siab tshaj ntawm HPG-axis ntsig txog cov noob.

AR yog lub hom phiaj ntawm ECH.

Txhawm rau kom paub meej txog kev sib raug zoo ntawm ECH thiab AR, ECH-Ovalbumin (ECH-OVA) tau ua tiav tiav (Fig. 5A, B). Qhov hnyav molecular ntawm ECH-OVA yog qhov loj dua OVA, yog li lawv muaj qhov sib txawv ntawm kev txav mus los (Fig. 5A), thiab lub siab absorbance ntawm ECH-OVA yog nruab nrab ntawm ECH thiab OVA (Fig. 5B),

qhia tias ECH tau ua tiav nrog OVA. Indirect ELISA cov txiaj ntsig (Daim duab 5C), qhia tias cov tshuaj tiv thaiv AR antibody muaj peev xwm kuaj tau AR protein nyob hauv qhov dej uas muaj ECH-OVA (qhia los ntawm anti-OVA antibody) qhia tias ECH tuaj yeem khi rau AR protein.

ECH ua ke nrog AR hnab tshos ntawm Met-894 thiab Val-713.

Compound ECH tau docked mus rau AF2 qhov chaw nyob rau saum npoo ntawm tib neeg AR, thiab theoretical binding hom ntawm ECH thiab AR tau qhia hauv daim duab 5D. Compound ECH tau txais kev sib cog lus sib txuas ntawm lub hnab tshos ntawm tib neeg AR. Te ob pawg phenyl ntawm ECH khi ntawm hydrophobic domain ntawm AR hnab tshos thiab khaws cia ze hydrophobic tiv tauj nrog cov residues Leu-712, Val-716, Met-734, Ile-737 , Tau ntsib -894, thiab Ile-898, whereas ob sab ntawm ECH tau muab tso rau ntawm qhov nkag ntawm lub hnab tshos thiab tsuas yog ob peb hu. Kev soj ntsuam ntxaws pom tau tias ob qho tib si Met-894 (cov nyiaj tshuav ntev: 2.4Å) thiab Val-713 (cov nyiaj tshuav ntev: 2.7Å) tsim cov ntawv cog lus hydrogen nrog cov hydroxy1 pawg ntawm ECH, uas yog qhov tseem ceeb ntawm ECH. thiab human AR.

Cistanche echinacoside tuaj yeemtxhim kho daim siab ua haujlwmthiabtxhim khu testosterone

ECH attenuates BPA-induced reproductive puas tsuaj.

Txhawm rau tshawb xyuas cov txiaj ntsig ntawm ECH ntawm kev ua me nyuam kev puas tsuaj, cov phev zoo thiab qib ntawm cov tshuaj hormones poj niam txiv neej thiab cov steroidogenic enzymes raug tshuaj xyuas hauv BPA-induced nas. Raws li pom nyob rau hauv daim duab 6, BPA kev kho mob txo cov phev suav thiab phev motility los ntawm 26.5 feem pua ​​thiab 39.2 feem pua. Kev tswj hwm ECH tiv thaiv qhov txo qis ntawm cov phev suav thiab phev txav los ntawm 35.5 feem pua ​​​​thiab 30.1 feem pua, raws li. Kev tswj hwm BPA kuj ua rau muaj qhov txo qis hauv LH thiab T secretion, thiab ECH tau nce qib ntawm LH thiab T los ntawm 24.1 feem pua ​​thiab 18.3 feem pua, raws li piv rau BPA kev kho mob ib leeg (Fig. 6A, B). Cov qib mRNA ntawm Star, CYP17A1, 3 -HSD, thiab 17 -HSD hauv BPA-ECH kev kho mob tau nce ntau dua piv rau kev kho BPA (Fig. 6C, D).