PartⅠ Kev Txhim Kho hauv Cov Ntshav Urea Nitrogen Thiab Serum Creatinine Siv Kev Loj Hlob Ntawm Cordyceps Militaris

Abstract

Mob raum mob (CKD) yog ib qho teeb meem tseem ceeb ntawm kev noj qab haus huv rau pej xeem nrog lub nra nyiaj txiag loj rau cov neeg mob thiab tib neeg thoob ntiaj teb. Hmoov tsis zoo, tam sim no tsis muaj cov kev kho mob zoo los tiv thaiv lossis ncua kev loj hlob ntawm cov kab mob raum kawg (ESRD). Kev coj noj coj ua hauv Suav teb tau pom tias cov tub rog Cordyceps (C. tub rog) mycelia muaj ntau yam tshuaj muaj txiaj ntsig zoo, suav nrog tshuaj tua kab mob, tshuaj tiv thaiv kab mob, thiab hepatoprotection. Txawm li cas los xij, cov txiaj ntsig ntawm mycelial C. tub rog ntawm CKD tseem tsis paub meej. Cov txheej txheem. Ntawm no, peb tau tshawb xyuas qhov cuam tshuam ntawm C. militaris mycelia ntawm nas nrog CKD siv plaub hom xov xwm: HKS, HKS nrog cov vitamin A (HKS ntxiv rau A), CM, thiab CM nrog cov vitamin A (CM ntxiv rau A). Cov txiaj ntsig. .e cov txiaj ntsig ntawm hnub 10 tau qhia tias cov ntshav urea nitrogen (BUN) tau qis dua hauv HKS (41 feem pua), HKS ntxiv rau A (41 feem pua), thiab CM ntxiv rau A (34 feem pua) pawg piv nrog cov nyob hauv Cov pab pawg tswj tau sib xws (cov nas nephrectomy). .e theem ntawm cov ntshav creatinine hauv HKS ntxiv rau pawg A tau txo qis los ntawm 35 feem pua ​​​​ntawm hnub 10, thaum cov qib hauv HKS thiab CM ntxiv rau pawg A tau txo qis los ntawm 14 feem pua ​​​​thiab 13 feem pua, raws li, hnub 30. Ua ke, qhov no yog thawj daim ntawv tshaj tawm siv plaub qhov xov xwm tshiab (HKS, HKS ntxiv rau A, CM, thiab CM ntxiv rau A nruab nrab) rau C. militaris mycelia. Txhua qhov nruab nrab ntawm mycelial C. tub rog ntawm CKD nthuav tawm cov txiaj ntsig tshwj xeeb ntawm BUN, ntshav creatinine, lub cev hnyav, tag nrho cov protein, thiab uric acid. Cov lus xaus. Ua ke, qhov no yog thawj daim ntawv tshaj tawm uas siv plaub qhov xov xwm tshiab (HKS, HKS ntxiv rau A, CM, thiab CM ntxiv rau A nruab nrab) rau C. militaris mycelia. Txhua qhov nruab nrab ntawm mycelial C. tub rog ntawm CKD nthuav tawm cov teebmeem tshwj xeeb ntawm BUN, ntshav creatinine, lub cev hnyav, tag nrho cov protein, thiab uric acid. Peb xaus lus tias kev kho mob nrog C. militaris mycelia kab lis kev cai hauv HKS lossis CM ntxiv rau A nruab nrab tuaj yeem tiv thaiv kev ua haujlwm tsis zoo ntawm lub raum hauv nas nrog CKD.

Ntsiab lus

Mob raum mob (CKD), Cordyceps Sinensis, Mycelium, Ntshav urea nitrogen (BUN), Serum creatinine,Cov txiaj ntsig Cistanche.

Nyem qhov no kom tau txaiscov teebmeem ntawm Cistanche

Taw qhia

Mob raum mob (CKD) yog ib qho teeb meem kev noj qab haus huv thoob ntiaj teb [1]. CKD yog ib lo lus dav dav rau cov kab mob heterogeneous cuam tshuam rau lub raum qauv thiab kev ua haujlwm [2]. Cov neeg mob nrog CKD muaj kev pheej hmoo siab ntawm cov kab mob raum kawg (ESRD) [3, 4]. Cov kws kho mob tam sim no tsom rau cov teeb meem cuam tshuam nrog kev kho CKD, xws li hyperlipidemia, ntshav qab zib, thiab ntshav siab ntau dua li tshwj xeeb kho CKD nws tus kheej [5–8], ua rau kev kawm txog kev tiv thaiv lub raum ib qho kev kho mob tshiab. Cov kev tshawb fawb yav dhau los tau tsom mus rau homeodomain-interacting protein kinase 2 (HIPK2) vim tias nws yog ib qho transcriptional regulator ntawm gene qhia koom nrog hauv tubular raug mob thiab fibrosis [9, 10]; Txawm li cas los xij, tshwj xeeb HIPK2 inhibitors tsis muaj muag. Tsis tas li ntawd, thaiv cov renin-angiotensin-aldosterone system tau pom tias yuav txo tau ob qho kev pheej hmoo ntawm hyperkalemia kev loj hlob thiab tus nqi rov qab [11, 12]. Txawm hais tias cov tshuaj renin-inhibiting aliskiren yav dhau los tau muab coj los ua ke nrog angiotensin-hloov enzyme inhibitors lossis angiotensin receptor blockers tsis tu ncua, tam sim no tau siv ntau qhov kev saib xyuas vim tias nws cov kev mob tshwm sim hnyav [13].

Nws tau raug tshaj tawm tias muaj feem cuam tshuam rau cov kab mob raum ntev yog hnub nyoog, haiv neeg, rog rog, ntshav qab zib, qhov hnyav yug me nyuam, ntshav siab, thiab tsev neeg keeb kwm [14]. .e kev pheej hmoo ntawm CKD morbidity thiab kev tuag tseem ceeb heev, nrog rau cov neeg mob CKD feem ntau tau txais kev kho lub raum, xws li kev lim ntshav thiab hloov lub raum. .peb, kev kho tshiab yuav tsum tau tsim. Nyob rau hauv xyoo tas los no, kev kho tshuaj ntsuab tau muab lwm txoj kev kho mob rau CKD [15, 16]. Tsis tas li ntawd, ntau hom kev tshawb fawb tau nthuav tawm tias cov tshuaj suav tshuaj ntsuab tsim nyog muaj txiaj ntsig zoo rau CKD, uas tuaj yeem txo qis kev pheej hmoo ntawm ESRD hauv cov neeg mob CKD thiab txhim kho cov neeg mob uas muaj CKD mus ntev. Piv txwv li, kev ua tau zoo ntawm ntau yam tshuaj ntsuab, suav nrog Radix Astragali, Rheum officinale, Panax ginseng, thiab Lycopus lucidus, ntawm cov kab mob raum tau tshawb xyuas [18–21]. Qee cov tshuaj ntsuab tau pom tias muaj txiaj ntsig zoo hauv kev txo qis proteinuria lossis nce ntshav albumin. Lwm tus, txawm li cas los xij, muaj cov khoom xyaw tshuaj lom, xws li aristolochic acid lossis cov hlau hnyav, uas tuaj yeem cuam tshuam rau lub raum ua haujlwm thiab ua rau nephropathy [22]. Txawm hais tias lawv tau siv ntau zaus hauv qee lub tebchaws tsim kho, cov lus ceeb toom ntawm lawv cov kev ua tau zoo tseem muaj teeb meem. .erefore, tsim cov khoom siv tau zoo los ntawm cov khoom siv ntuj tsim los kho CKD yog qhov kev txhawj xeeb ceev.

Cordyceps belongs rau tsev neeg fungus thiab yog ib hom tshuaj suav tshuaj uas muaj kab mob cab kab mob loj hlob thiab maj mam tig mus rau hauv lub cev loj hlob. Cordyceps sinensis (C. sinensis) thiab Cordyceps tub rog (C. tub rog) yog ob hom Cordyceps paub zoo. Tau ntau xyoo, C. sinensis tau siv los kho kev qaug zog, lub raum thiab lub ntsws ua haujlwm tsis zoo, hyperglycemia, hyperlipidemia, thiab arrhythmia [23], thiab nws cov teebmeem hauv CKD cov neeg mob thiab raum hloov pauv tau raug kawm [24–26]. Feem ntau, C. militaris yog qhov zoo rau kev tsim khoom loj [27] thiab nthuav tawm ob qho tib si sib txawv thiab tshwj xeeb cov khoom siv tshuaj [28–30]. Tsis tas li ntawd, ib txoj kev tshawb fawb yav dhau los ua los ntawm peb pab pawg tau tshawb xyuas cov haujlwm anticancer ntawm C. militaris thiab pom tau tias nws cov mycelial fermentation tuaj yeem tswj cov mitogen-activated protein kinases (MAPK) teeb liab txoj hauv kev kom nres lub voj voog ntawm tes, txhawb nqa chromosomal DNA tawg, thiab thaum kawg. ua rau ob qho tib si apoptotic thiab autophagic tuag ntawm kab lis kev cai glioblastoma hlwb [31]. .erefore, C. militaris yuav ua hauj lwm rau nws cov teebmeem renoprotective hauv txoj kev tshawb no.

Mycelial C. militaris tsis ntev los no tau tsim los ua khoom noj khoom haus nrov hauv Asia. Extracts los ntawm C. militaris txiv hmab txiv ntoo lub cev tuaj yeem ncua kev loj hlob ntawm lub raum malfunction induced los ntawm subtotal nephrectomy [32]. Kev tshawb fawb txog cov nyhuv ntawm C. militaris mycelia ntawm CKD, txawm li cas los xij, tsis tshua muaj. .is txoj kev tshawb fawb yuav sim peb qhov kev xav tias mycelial C. militaris tuaj yeem tiv thaiv kev ua haujlwm tsis zoo ntawm lub raum hauv nas nrog CKD. Plaub hom xov xwm yuav raug tsim thiab ua haujlwm los tsim cov tub rog mycelial C. tub rog: (i) HKS, (ii) HKS ntxiv rau A, (iii) CM, thiab (iv) CM ntxiv A. Rau 30 hnub, cov nas hauv pab pawg kho mob yuav tau txais kev tswj hwm txhua hnub los ntawm qhov ncauj gavage ntawm C. militaris mycelia kab lis kev cai nyob rau hauv ib qho ntawm plaub qhov sib txawv ntawm cov xov xwm, whereas cov nas nyob rau hauv sham tswj (C) thiab nephrectomy tswj (Nx) pawg yuav tau txais cov dej distilled. .ese kev ntsuam xyuas yuav tsum tau qhia meej txog kev nyab xeeb ntawm mycelial C. militaris ua zaub mov ua haujlwm.

Cistanche tubulosa

Cov ntaub ntawv thiab cov txheej txheem

1. Khoom siv.

C. militaris mycelia (BCRC 32219) tau yuav los ntawm Bioresource Collection thiab Research Center ntawm Food Industry Research and Development Institute (Hsinchu, Taiwan). Glucose tau yuav los ntawm JT Baker (EU). Malt extract, peptone, thiab poov xab extract tau yuav los ntawm Becton Dickinson (Franklin Lakes, NJ, USA). Vitamin A tau muas los ntawm Sigma-Aldrich (St. Louis, MO, USA) thiab agar tau yuav los ntawm High Standard Enterprise Co., Ltd. (Taiwan), feem. Cov khoom siv tshuaj biochemical kuaj xyuas lub raum ua haujlwm tau txais los ntawm Arkray (Kyoto, Nyiv).

2. Fungus Media Kev Npaj.

Mycelial C. militaris yog incubated nyob rau hauv plaub hom xov xwm, namely CM, CM ntxiv rau A, HKS, thiab HKS ntxiv A. thiab 2 feem pua ​​​​qabzib [33]. .e HKS nruab nrab yog ib qho kev hloov kho ntawm qhov nruab nrab tsim los ntawm Prof. Huang Keng-Shiang thiab muaj 2 feem pua ​​​​malt extract, 2 feem pua ​​agar, 0.2 feem pua ​​peptone, thiab 0.2 feem pua ​​poov xab extract . .e CM thiab HKS xov xwm sib txawv nyob rau hauv cov concentrations ntawm peptone thiab poov xab extract. .e HKS ntxiv rau A thiab CM ntxiv rau A xov xwm txhua tus muaj 1 feem pua ​​​​vitamin A ntxiv rau hauv lub hauv paus nruab nrab. A 0.5 × 0.5 cm square ntawm C. militaris mycelium raug txiav thiab hloov mus rau txhua lub phaj thiab incubated ntawm 25 degree . Tom qab C. militaris tau coj mus rau hauv cov xov xwm sib txawv rau 30 hnub, cov kab mob mycelia tau ua tib zoo khawb tawm ntawm qhov chaw ntawm cov khoom nruab nrab siv riam. .e sau cov fungal mycelia hmoov tau khov-qhuav (EYELA FDU-1100) hauv lub tshuab nqus tsev ntawm −54 degree rau 48 teev. .e khov-qhuav C. militaris mycelia hmoov tau khaws cia ntawm −20 degree kom txog thaum siv.

3. CKD Mice Model Tsim.

.e tsib-sixth nephrectomy yog txoj kev tsim kho mob ntev tshaj plaws uas ua rau lub raum tsis ua haujlwm tom qab poob lub raum loj. .e CKD nas tau tsim los tom qab ob-kauj ruam, tsib-rau-thib nephrectomy raws li tau piav qhia yav dhau los [34, 35]. Luv luv, lub raum sab laug tau raug nthuav tawm thiab txiav rau ntawm lub sab sauv thiab qis ib feem peb ntawm tus ncej. .e 2/3 ntawm cov ceg ntoo ntawm lub raum cov hlab ntsha ntawm lub raum sab laug tau ligated thiab tom qab ntawd ua raws li txoj cai nephrectomy tom qab ib lub lim tiam. Tsiaj txhu raug xa rov qab mus rau lawv lub tawb tom qab kev phais tsawg kawg ob lub lis piam ua ntej uremia raug ntxias. .e cov txheej txheem tau txais kev pom zoo los ntawm Pawg Saib Xyuas Tsiaj Tsiaj thiab Siv Khoom ntawm I-Shou University (Kev Pom Zoo No: IACUC-ISU- 101025).

4. Cov txheej txheem sim.

Cov hmoov qhuav qhuav ntawm C. militaris mycelia tau tsau rau hauv dej distilled (10 mg / mL) ntawm chav tsev kub. Ob lub lis piam tom qab qhov thib ob nephrectomy, cov nas tau muab faib ua pawg kho mob (n=4 rau 6 ib pawg) thiab kho nrog ib qho ntawm plaub hom C. militaris mycelia daws, incubated hauv HKS, HKS ntxiv rau A. , CM, lossis CM plus A nruab nrab, los ntawm qhov ncauj gavage rau 30 hnub sib law liag. Pawg sham thiab Nx nas tau tswj hwm qhov sib npaug ntawm cov dej distilled yam tsis muaj C. militaris mycelia. Lub cev hnyav tau ntsuas thiab kuaj ntshav tau sau rau hnub 1, 10, thiab 30, feem. Thaum kawg ntawm lub sijhawm kho, cov nas tau txi nrog CO2. .e ob lub raum raug muab tshem tawm thiab ntxuav nrog phosphate-buffered saline thiab muab tso rau hauv 10 feem pua ​​​​ntxhiab tsw buffered formalin rau tom ntej histological ua. Cov ntshav kuaj tau sau los ntawm periorbital venous sinus. Ntshav urea nitrogen (BUN), ntshav creatinine, tag nrho cov protein, thiab uric acid tau ntsuas los ntawm cov khoom lag luam uas siv cov khoom siv tshuaj biochemical automated (SPOTCHEM EZ SP-4430) raws li cov chaw tsim khoom cov lus qhia (Arkary, Inc., Kyoto, Nyiv).

Standardized Cistanche

5. Tsiaj noj.

Lub koom haum ntawm Cancer Research txiv neej nas (kwv yees li 30 g) tau muab los ntawm BioLASCO Taiwan Co., Ltd. thiab khaws cia rau hauv lub tawb txheem ntawm qhov kub thiab txias ntawm 22 ± 1 degree nrog rau 12- teev lub teeb-tsaus voj voog. Tsiaj txhu tau noj nrog nas chow li niaj zaus thiab kais dej ad libitum. .e tsiaj txhu siv nyob rau hauv txoj kev tshawb no tau nyob hauv tsev thiab saib xyuas los ntawm NIH Phau Ntawv Qhia Txog Kev Saib Xyuas thiab Siv Cov Tsiaj Looj Mem.

6. Kev tshuaj ntsuam biochemical ntawm cov ntshav kuaj.

Cov ntshav kuaj tau sau los ntawm periorbital venous sinus. Cov qauv ntshav tau centrifuged ntawm 12, 000 rpm rau 10 min ntawm 4 degree thiab khaws cia ntawm −20 degree ua ntej kev tshuaj xyuas. Ntshav urea nitrogen (BUN), ntshav creatinine, tag nrho cov protein, thiab uric acid tau ntsuas los ntawm cov khoom siv muag khoom siv cov khoom siv biochemical automated (SPOTCHEM EZ SP-4430) raws li cov chaw tsim khoom cov lus qhia (Arkary, Inc., Kyoto, Nyiv).

7. Lub raum Histological Analysis.

Cov nas lub raum tau muab tso rau hauv paraffin blocks, dissected rau hauv 3-μm-thick seem, thiab ua tiav nrog Harry's hematoxylin-eosin (HE) stain tom qab tus txheej txheem txheej txheem. .e lub raum glomeruli thiab tubules raug tshuaj xyuas thiab thaij duab rau yav tom ntej. Cov duab raug ntes tau siv lub koob yees duab xim (VKC150, Hitachi, Tokyo, Nyiv) txuas nrog lub tshuab ntsuas (DP72, Olympus, Center Valley, PA, USA) thiab tsom mus rau qhov muag tsis pom los ntawm tus kws paub txog kab mob. .e txhais tau hais tias glomerular cross-sectional cheeb tsam tau los ntawm kev xam qhov nruab nrab thaj tsam ntawm kwv yees li 8 mus rau 15 tus neeg glomeruli siv qhov program Image J.

8. Kev txheeb xyuas qhov tseeb.

Tag nrho cov ntaub ntawv raug nthuav tawm raws li qhov ntsuas- ± tus qauv kev ua yuam kev ntawm qhov nruab nrab (n=4 rau 6 ib pawg). Cov ntaub ntawv raug tshuaj xyuas siv ob txoj kev tsom xam ntawm qhov sib txawv raws li Bonferroni post hoc xeem (SigmaPlot version 10.0, San Jose, CA, USA). p <0.05 tau suav tias yog qhov tseem ceeb.

Cistanche extract

Cov ntaub ntawv

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Chih-Hui Yang, 1,2,3Wen-Shuo Kuo,4 Jun-ShengWang,2Yi-Ping Hsiang,3,5Yu-Mei Lin,1,6 Yi-Ting Wang, 1,6 Fan-Hsuan Tsai,6 Chun- Ting Lee, 6,7 Jiun-Hua Chou, 1 Huei-Ya Chang, 1 Lung-Shuo Wang, 6,8 Shu-Chi Wang, 6,9 thiab Keng-Shiang Huang 6

1 Department of Biological Science thiab Technology, I-Shou University, Kaohsiung, Taiwan

2 Taiwan Instrument Research Institute, National Applied Research Laboratories, Hsinchu, Taiwan

3 Pharmacy Department of E-Da Tsev Kho Mob, Kaohsiung, Taiwan

4 Lub Tsev Kawm Ntawv ntawm Chemistry thiab Materials Science, Nanjing University of Information Science thiab Technology, Nanjing, Suav

5 Department of Biotechnology thiab Chemical Engineering, I-Shou University, Kaohsiung, Taiwan

6 Lub Tsev Kawm Ntawv ntawm Suav Tshuaj rau Kev Kawm Qib Siab, I-Shou University, Kaohsiung, Taiwan

7 Amulette Suav Tshuaj Kho Mob, Taipei, Taiwan

8 Department of Chinese Medicine, Sin-Lau Tsev Kho Mob, Tainan, Taiwan

9 Lub Tsev Kawm Ntawv Tshuaj rau Cov Tub Ntxhais Kawm Thoob Ntiaj Teb, I-Shou University, Kaohsiung, Taiwan