Hypoglycemic Thiab Hypolipidemic Los Ntawm Tag Nrho Glycosides Ntawm Cistanche Tubulosa hauv Kev Noj Qab Haus Huv / streptozotocin-induced Diabetic nas

Hu rau: Audrey Huaudrey.hu@wecistanche.com

Kuiniu Zhu, et al

TSAB NTAWV

Ethnopharmacological qhov tseeb: Cistanche tubulosa(Schrenk) R. Wight (Orobanchaceae) yog ib qho tshuaj uas nquag sau ua ntej hauv ntau cov tshuaj ntsuab tshuaj uas siv los kho ntshav qab zib hauv Suav teb. Nyob rau hauv cov kev tshawb fawb tsis ntev los no, kev ua haujlwm antidiabetic ntawmCistanchetubulosaextracts tau lees paub. Txawm li cas los xij, tsis muaj kev tshawb fawb tau qhia txog tag nrho glycosides ntawmTubulosa(TGCT). Lub hom phiaj ntawm txoj kev tshawb no: Txoj kev tshawb fawb tam sim no tsom rau kev tshawb xyuas cov teebmeem hypoglycemic thiab hypolipidemic ntawm TGCT thiab cov txheej txheem muaj peev xwm hauv kev noj zaub mov / streptozotocin (STZ) vim cov nas mob ntshav qab zib, thiab txhawm rau txheeb xyuas cov ntsiab lus tseem ceeb ntawm TGCT (tag nrho glycosides ntawmCistanchetubulosa).

Cov ntaub ntawv thiab cov txheej txheem:Cov ntsiab lus tseem ceeb ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa)tau tus cwj pwm los ntawm HPLC/Q-TOF-MS thiab cov kev ntsuas ntsuas tau ua nrog HPLC-DAD. Cov nas mob ntshav qab zib hom 2 tau tshwm sim los ntawm kev noj zaub mov muaj rog ntau (HFSD) thiab ib qho kev txhaj tshuaj ntawm STZ (30 mg / kg). TGCT (50 mg / kg, 100 mg / kg thiab 200 mg / kg) lossis metformin (200 mg / kg) tau muab tshuaj rau qhov ncauj rau 6 lub lis piam. Lub cev hnyav thiab calorie kom tsawg tau saib xyuas thoob plaws qhov kev sim. Fasting plasma glucose (FPG), qhov ncauj qhov ncauj glucose tolerance test (OGTT), thaj tsam hauv qab ntawm cov piam thaj (AUC-G), glycosylated hemoglobin (HbA1c), yoo mov insulin, ntshav C-peptide, glycogen cov ntsiab lus, thiab insulin rhiab heev index. kuaj. Cov theem ntawm phosphorylated protein kinase B thiab phosphorylated glycogen synthase kinase 3, cov dej num ntawm hexokinase thiab pyruvate kinase tau soj ntsuam. Lub caij no, cov kev hloov pauv hauv cov ntshav lipid profile, superoxide dismutase, glutathione peroxidase, malondialdehyde, thiab inflammatory yam raug ntsuas. Histological ntawm pancreas kuj tau soj ntsuam los ntawm hematoxylin-eosin stain.

Cov txiaj ntsig:Peb qhov kev tshawb nrhiav tau pom tias muaj phenylethanoid glycosides (PhGs): echinacoside (500.19 ± 11.52 mg / g), acteoside (19.13 ± 1.44 mg / g) thiab isoacteoside (141.82 ± 5.78 mg / g) hauv TGCT(tag nrho glycosides ntawmCistanchetubulosa). Kev ntsuam xyuas kws tshuaj qhia tau hais tias TGCT thim rov qab STZ-vim qhov hnyav poob (11.1 feem pua ​​, 200 mg/kg); txo FPG (56.4 feem pua, 200 mg/kg) thiab HbA1c (37.4 feem pua, 200 mg/kg); ameliorated OGTT, AUC-G, thiab insulin rhiab heev; nce glycogen cov ntsiab lus (40.8 feem pua ​​​​ntawm daim siab thiab 52.6 feem pua ​​​​ntawm cov leeg, 200 mg / kg) thiab cov dej num ntawm carbohydrate metabolizing enzymes; tswj lipid profile hloov pauv thiab cov haujlwm ntawm cov enzymes antioxidant; txo cov ntshav cov cim ntawm oxidative kev nyuaj siab thiab o nyob rau hauv ib koob tshuaj raws li (p <>

Cov lus xaus:Txoj kev tshawb no tau lees paub tias TGCT(tag nrho glycosides ntawmCistanchetubulosa) yog ib qho khoom noj muaj txiaj ntsig zoo rau ameliorating hyperglycemia thiab hyperlipidemia hauv kev noj zaub mov / STZ-induced ntshav qab zib nas, uas tej zaum yuav raug ntaus nqi rau cov dej num ntawm TGCT.(tag nrho glycosides ntawmCistanchetubulosa)ntawm inhibitions ntawm oxidative kev nyuaj siab thiab o.

Ntsiab lus:Tag nrho glycosides ntawmCistanche tubulosaHom 2 mob ntshav qab zib mellitus Hypoglycemic Hypolipidemic Anti-oxidant Anti-inflammatory

1. Taw qhia

Mob ntshav qab zib mellitus yog suav tias yog kab mob metabolic, uas yog tus yam ntxwv ntawm hyperglycemia uas tshwm sim los ntawm kev tsim cov tshuaj insulin tsis txaus lossis / thiab insulin tsis kam (IR) (American Diabetes Association, 2019). Mob hyperglycemia yog txuam nrog ntau yam teeb meem loj xws li nephropathy, retinopathy, neuropathy, thiab teeb meem plawv (Ekoe, 2019). Tus naj npawb ntawm cov neeg mob ntshav qab zib hauv ntiaj teb xyoo 2019 kwv yees yog 463 lab (9.3 feem pua), nce mus rau 578 lab (10.2 feem pua) los ntawm 2030 thiab 700 lab (10.9 feem pua) los ntawm 2045 (Saeedi li al., 2019). Ntau cov tshuaj xws li metformin (Met, txhawb kev tsim cov kab mob siab glycogen), insulin (txo kev tsim cov piam thaj thiab nce kev siv cov piam thaj), thiab sulfonylureas (txhim kho cov kab mob pancreatic islet cell kom zais cov tshuaj insulin) zoo txo ​​qis glycemia. Txawm li cas los xij, ntau qhov kev tsis txaus siab tsis zoo (xws li qhov hnyav nce, hypoglycemia, IR, thiab edema) tau txwv lawv siv (Moller, 2001). Yog li ntawd, ntau tus kws tshawb fawb tau tshawb nrhiav cov tshuaj lom neeg lom neeg los ntawm cov tshuaj cog qoob loo rau kev kho mob ntshav qab zib xyoo dhau los (Kasangana li al., 2019; Liu et al., 2020).

Cistanchetubulosa(Schrenk) R. Wight (Orobanchaceae) tau siv dav hauv cov tshuaj suav tshuaj, uas feem ntau tau sau tseg hauv cov qauv tshuaj kho mob raum tsis txaus, poj niam muaj menyuam, thiab ntshav qab zib mellitus (Li et al., 2016; Han li al., 2017). ; Su et al., 2017). Cov kev tshawb fawb tsis ntev los no tau tshaj tawm tias aqueous extract ntawmCistanchetubulosapom cov teebmeem hypoglycemic thiab hypolipidemic hauv db / db nas nrog hom 2 mob ntshav qab zib mellitus (T2DM) (Xiong li al., 2013), thiab ameliorated ntshav qabzib qib, IR, thiab lipid peroxidation hauv streptozotocin (STZ)-nicotinamide-induced Diabetic rats Kong et al., 2018). Phenylethanoid glycosides (PhGs) yog cov khoom tseem ceeb ntawmCistanchetubulosa(Morikawa et al., 2014), uas tau nthuav tawm ntau yam kev ua ub no xws li tshuaj tiv thaiv oxidation (Xue et al., 2017) thiab tshuaj tiv thaiv kab mob cancer (Fu et al., 2019). Ntxiv mus, PhGs cuam tshuam qhov nce ntawm postprandial ntshav qabzib qib hauv cov hmoov txhuv nplej siab loaded nas (Morikawa li al., 2014), suppressed cov sodium-dependent qabzib cotransporter 1-cov piam thaj hauv nruab nrab ntawm cov plab hnyuv siab raum (Shimada li al., 2017), thiab inhibited qhov kev ua ntawm aldose reductase hauv nas lo ntsiab muag (Morikawa li al., 2019). Txawm li cas los xij, tsis muaj kev tshawb fawb yav dhau los tau tshawb xyuas qhov kev tiv thaiv hyperglycemic ntawm tag nrho glycosides ntawmCistanchetubulosa(TGCT).

Hauv kev tshawb fawb tam sim no, cov tshuaj tiv thaiv kab mob ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa)tau raug soj ntsuam nyob rau hauv cov zaub mov muaj roj siab-sucrose (HFSD) thiab STZ-induced ntshav qab zib nas. Tsis tas li ntawd, cov tshuaj tua kab mob antioxidant thiab tshuaj tiv thaiv kab mob kuj tau tshawb xyuas kom nkag siab txog lub peev xwm ntawm TGCT.

2 Cov ntaub ntawv thiab cov txheej txheem

2.1. Tshuaj thiab reagents

STZ tau yuav los ntawm Sigma-Aldrich Corp. (Saint Louis, USA). Kev sib ntsib tau txais los ntawm Sino-American Shanghai Squibb Pharma. Cov khoom siv ELISA ntawm insulin, C-peptide tau muab los ntawm Elabscience Biotechnology Co., Ltd (Wuhan, Suav). Cov khoom siv ELISA cov piam thaj, glycosylated hemoglobin (HbA1c), tag nrho cov roj (TC), triacylglycerol (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), superoxide dismutase (SOD) , glutathione peroxidase (GSH-Px), malondialdehyde (MDA), qog necrosis factor- (TNF), interleukin 1 (IL-1), interleukin 6 (IL-6) tau yuav los ntawm Nanjing Jiancheng Bioengineering Institute . Cov khoom siv ELISA ntawm phosphorylated protein kinase B (p-PKB) thiab phosphorylated glycogen synthase kinase 3 (p-GSK3) tau txais los ntawm Shanghai Enzyme-linked Biotechnology Co., Ltd. Echinacoside (purity Ntau dua lossis sib npaug li 98 feem pua) thiab acteoside (purity Ntau dua lossis sib npaug li 97 feem pua) tau yuav los ntawm National Institutes for Food and Drug Control. Isoacteoside (purity Ntau dua lossis sib npaug li 98 feem pua) yog muab los ntawm Chengdu Yuav Tsum Biotechnology Co., Ltd.

2.2. Cov peev txheej cog thiab kev npaj ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa)

Qhuav succulent qia ntawmCistanchetubulosatau yuav los ntawm Bozhou Yihongtang Pharmaceutical Co., Ltd., (Anhui, Tuam Tshoj) thiab txheeb xyuas los ntawm Dr. Puyang Gong ntawm Pharmaceutical Botany Department, Southwest Minzu University. Daim ntawv pov thawj daim ntawv pov thawj (No. 20161103) raug crushed rau hauv cov hmoov thiab muab tso rau hauv herbarium ntawm Jiangsu Kanion Pharmaceutical Co., Ltd. Raws li SFDA lub teb chaws tshuaj tus qauv "Congrong Zonggan Jiaonang" (YBZ07482005-2011Z), lub tshuaj crude (1 kg) tau muab rho tawm nrog dej rau peb zaug tom qab soaking rau 1 h. Tom qab filtration, lub filtrate yog concentrated nyob rau hauv lub siab txo, ntxiv cawv rau cov concentrated tov kom txog thaum lub ethanol concentration mus txog 60 feem pua. Cov kua supernatant yog concentrated kom tsis muaj cawv saj thiab ces yuav purified los ntawm macroporous resin. Ua ntej, dej eluent thiab 40 feem pua ​​​​ethanol eluent tau sau ua tiav rau kev siv tom qab. Qhov thib ob, cov dej eluent tau rov ua dua rau hauv macroporous resin thiab eluted nrog dej. Cov dej eluent tau muab pov tseg. Thib peb, elution nrog 40 feem pua ​​​​ethanol thiab cov eluent tau sau rau tom qab siv. Thaum kawg, 40 feem pua ​​​​ethanol eluent tau muab tso ua ke thiab concentrated los ntawm rotary evaporator, ces cov tshuaj tau qhuav los ntawm cov tshuaj tsuag kom qhuav. Txog 60 g xim av tau txais (piv txwv li TGCT). Lub purity ntawm TGCT tau kuaj pom raws li tus qauv (YBZ07482005-2011Z), uas tau mus txog 853 mg / g. TGCT(tag nrho glycosides ntawmCistanchetubulosa)({{0}}}.21 mg) thiab peb cov qauv sib xyaw (echinacoside: 131.3 ug, acteoside: 4.2 ug, isoacteoside: 39.4 ug) raws li yaj hauv 1 mL methanol: dej (50/50, v / v), thiab tom qab ntawd lim los ntawm 0.45 μm membrane ua ntej txhaj tshuaj.

2.3. Kev soj ntsuam zoo ntawm TGCT los ntawm HPLC/Q-TOF-MS

HPLC system tau cuam tshuam nrog Agilent 6538 Q-TOF-MS (Agilent Corp, USA) nruab nrog electrospray ionization. Kev tsom xam tau ua tiav ntawm Zorbax SB-C18 kem (15{{10}} mm × 4.6 mm, 5 μm). Cov theem txawb tau tsim los ntawm methanol-dej (muaj 0.1 feem pua ​​H3PO4). Tus nqi ntws yog 1.0 mL/min. Gradient elution cov kev pab cuam tau sau tseg raws li nram no: 0–20 min, 18 feem pua ​​– 28 feem pua ​​A; 20-50 min, 28% -32% A; 50–60 min, 32 feem pua ​​A; 60-70 feeb, 32% -50%. TOF-MS tau ua nyob rau hauv ob qho tib si zoo thiab tsis zoo ion hom tshaj m / z 100-3000 nyob rau hauv cov nram qab no tsis ua hauj lwm: capillary voltage 3500 V (ESI-) los yog 4000 V (ESI ntxiv); qhuav roj, 10.0 L / min; gas kub, 350 ◦C; nebulizer siab, 35 psi; skimmer voltage, 65V; fragment lossis voltage, 135 V; OCTRFV, 750 V. Tag nrho cov ntaub ntawv tau tswj los ntawm Cov Ntaub Ntawv Tau Txais rau TOF / Q-TOF Ver. B.03.01 and Qualitative Analysis Ver. B.03.01 (Agilent Technologies, USA).

2.4. Quantitative analysis of TGCT(tag nrho glycosides ntawmCistanchetubulosa)los ntawm HPLC-DAD

Kev sib cais tau ua tiav ntawm Zorbax SB-C18 kem (150 mm × 4.6 mm, 5 μm) hauv 50 min (1.0 mL / min). Chromatographic tej yam kev mob rau HPLC-DAD yog tib yam li kev soj ntsuam zoo. Lub ntim ntim, kem kub, thiab UV wavelength tau teem rau ntawm 5 μL, 30 ◦C, thiab 330 nm, feem.

2.5. Kev sim tsiaj

Txiv neej SD nas, hnyav 180 ± 20 g, tau yuav los ntawm Laboratory Tsiaj Center ntawm Nanjing Qinglongshan [Certificate No. SCXK (SU) 2017-0001]] thiab nyob hauv Chaw Saib Xyuas Tsiaj hauv Jiangsu Kanion Pharmaceutical Co., Ltd. (Jiangsu, Suav teb). Cov txheej txheem kev saib xyuas tsiaj thiab kev sim tau pom zoo los ntawm Pawg Saib Xyuas Tsiaj Tsiaj thiab Siv Khoom Siv, Huzhou Lub Tsev Haujlwm Saib Xyuas Khoom Noj thiab Tshuaj (pom zoo No. 19018), thiab tau ua raws li Cov Cai rau Tsiaj Kev sim ntawm Tuam Tshoj. Cov nas tau khaws cia nyob rau hauv qhov ntsuas kub (24 ± 2 ◦C) thiab av noo (50 ± 10 feem pua) nrog 12 h lub teeb thiab tsaus voj voog, acclimatized rau cov neeg nyob rau 7 hnub los ntawm ib txwm kuaj chow thiab dej ad libitum.

2.6. Induction ntawm ntshav qab zib

Cov nas nyob hauv pawg tswj hwm ib txwm muaj (NC, n {{0}}) tau noj nrog kev noj zaub mov ib txwm, thaum cov nas sim tau noj nrog HFSD (ib txwm noj zaub mov ntxiv nrog 20 feem pua ​​​​sucrose, 10 feem pua ​​​​lard, 2.5 feem pua ​​​​cov roj (cholesterol), thiab 1 feem pua ​​​​cholate, 3.95 kcal / g) rau 4 lub lis piam. Tom qab 12 teev yoo mov, nas tau txhaj tshuaj intraperitoneally nrog ib koob tshuaj STZ (30 mg / kg), uas yaj hauv qhov txias citrate tsis (0.1 M, pH 4.5) tam sim ua ntej siv. Hnub 8 ntawm STZ-txhaj tshuaj, cov ntshav tau coj los ntawm tus Tsov tus tw kawg los ntawm cov ntshav uas siv cov koob pov tseg, thiab yoo ntshav ntshav qabzib (FPG) tau txiav txim siab los ntawm lub portable glucometer (LifeScan, Inc. UK). Cov nas uas muaj cov tsos mob ntawm polyuria, polydipsia, thiab FPG Ntau dua lossis sib npaug li 11.1 mmol / L raug suav hais tias yog nas mob ntshav qab zib thiab muab faib ua tsib pawg (n =10).

Pab pawg I: NC, pub nrog 0.5 feem pua ​​​​sodium carboxyl methylcellulose (CMC Na, 10 mL/kg).

Pawg II: Kev tswj ntshav qab zib (DC), noj nrog 0.5 feem pua ​​CMC-Na (10 mL/kg).

Pawg III: TGCT-50, kho nrog TGCT (50 mg/kg).

Pab pawg Ⅳ: TGCT-100, kho nrog TGCT (100 mg/kg).

Pab pawg Ⅴ: TGCT-200, kho nrog TGCT (200 mg/kg).

Pab pawg Ⅵ: Tau ntsib -200, kho nrog Met (200 mg/kg).

Cov koob tshuaj siv nyob rau hauv txoj kev tshawb no tau raug xaiv raws li Suav Pharmacopeia (2015 tsab). Tag nrho cov pab pawg tau txais kev hais lus ib zaug ib hnub thiab txuas ntxiv tau txais lawv cov khoom noj rau lwm 6 lub lis piam.

2.7. Saib xyuas qhov xwm txheej ntawm nas

Cov tsos mob ntawm cov plaub hau, zis tso zis, thiab kev ciaj sia ntawm nas tau pom txhua hnub. Lub cev hnyav (BW) thiab calorie kom tsawg tau saib xyuas thoob plaws qhov kev sim. FPG raug soj ntsuam ntawm 0, 2nd, 4th thiab 6 lub lis piam tom qab kho nrog TGCT(tag nrho glycosides ntawmCistanchetubulosa).

2.8. Qhov ncauj Glucose Tolerance Test (OGTT)

OGTT tau ua rau cov nas yoo mov thaum hmo ntuj ntawm lub davhlau ya nyob twg ntawm tag nrho cov kev tshawb fawb. Tsuas yog 6 0 μL cov ntshav kuaj tau raug sau nrog cov kav dej capillary los ntawm orbital sinus (0 h), ces muab tshuaj nrog TGCT(tag nrho glycosides ntawmCistanchetubulosa)(50 mg/kg, 100 mg/kg, thiab 200 mg/kg) los yog Met (200 mg/kg) feem. Cov ntshav kuaj tau raug sau ntawm 0.5 h, 1 h, 2 h tom qab cov piam thaj load (2 g / kg). Cov nas tsuag tau muab tshuaj loog nrog isoflurane rau ob peb feeb ua ntej noj ntshav, thiab tom qab ntawd nyem tam sim kom tsis txhob los ntshav los ntawm hemostatic paj rwb. Txhua qhov kev sim tau ua tiav nrog kev saib xyuas zoo kom ntseeg tau tias muaj kev noj qab haus huv ntawm cov tsiaj. Plasma glucose concentrations tau txiav txim siab los ntawm cov khoom siv qabzib raws li txoj kev glucose oxidase peroxidase. Cov cheeb tsam hauv qab nkhaus ntawm qabzib (AUC-G) tau suav los xa mus rau cov ntaub ntawv (Shao li al., 2013).

2.9. Kev txiav txim siab ntawm kev yoo mov insulin (FINS) thiab insulin rhiab heev index (ISI)

Ib ntus ntawm 1 hnub tom qab OGTT, txhua tus nas nyob hauv qhov xwm txheej zoo yam tsis muaj tsos mob xws li qhov muag tsis pom kev thiab mob. Tom qab ntawd, lawv tau muab tshuaj loog nrog pentobarbital sodium (40 mg / kg, ip) tom qab yoo mov rau 12 teev thiab kuaj ntshav los ntawm lub plab aorta nrog thiab tsis muaj heparin rau kev kwv yees biochemical. Cov ntshav tau sau los ntawm cov ntshav kuaj (tsis muaj heparin) los ntawm centrifugation. FINS tau soj ntsuam los ntawm cov khoom siv ELISA. FPG tau txiav txim siab siv cov khoom siv coj mus muag raws li txoj kev qabzib peroxidase. ISI raug xam raws li tus qauv: ISI=1/[FINS (pmol/L) × FPG (mmol/L)] (Wang et al., 2013).

2.10. Kev kwv yees ntawm glycogen synthesis hauv daim siab thiab cov leeg

Lub siab thiab gastrocnemius cov leeg tau raug tshem tawm, yaug, hnyav, thiab khaws cia ntawm -70 ◦C. Glycogen hauv daim siab thiab cov leeg tau ntsuas los ntawm txoj kev anthrone raws li tau piav qhia dhau los (Ren li al., 2015). Cov ntsiab lus ntawm glycogen tau qhia raws li mg / g ntub qhov hnyav ntawm cov ntaub so ntswg. Cov dej num ntawm hexokinase (HK) thiab pyruvate kinase (PK) hauv lub siab tau txiav txim siab los ntawm cov khoom siv muag khoom raws li cov chaw tsim khoom cov lus qhia.

2.11. Biochemical tsom xam

HbA1c hauv cov ntshav tag nrho (nrog heparin) tau ntsuas los ntawm cov khoom siv kuaj mob. Serum C-peptide, p-PKB, p-GSK3, TC, TG, LDL-C, HDL-C, SOD, GSH-Px, MDA, TNF- , IL-6, thiab IL-1 tau sim siv cov khoom siv coj mus muag raws li cov chaw tsim khoom cov lus qhia.

2.12. Kev ntsuas histological ntawm pancreas

Pancreas cov ntaub so ntswg kuj raug tshem tawm, yaug, thiab kho hauv 10 feem pua ​​​​ntawm cov formalin, tom qab ntawd lub cev qhuav dej hauv gradient ethanol (75 feem pua, 85 feem pua, 95 feem pua, thiab 100 feem pua) thiab xylene (100 feem pua). Tom qab permeation, lawv tau embedded nyob rau hauv paraffin thiab txiav mus rau hauv 3 μm tuab seem nrog ib tug teb microtome. Cov ntaub so ntswg tau stained nrog hematoxylin-eosin (H&E) rau kev kuaj pom lub teeb (Chen li al., 2014).

2.13. Kev txheeb cais

Kev txheeb xyuas txheeb cais tau ua los ntawm SPSS version 16.0 software. Cov ntaub ntawv tau nthuav tawm los ntawm qhov nruab nrab ± SD. Kev sib piv ntawm cov pab pawg tau ua tiav siv ib-txoj kev ANOVA ua raws li Tukey qhov kev xeem thiab tus nqi ntawm p < 0.05="" raug="" coj="" los="" ua="" qhov="" tseem="">

3. Cov txiaj ntsig

3.1. Phytochemical tsom xam ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa)

Kev soj ntsuam zoo tau ua tiav thiab nthuav tawm hauv cov ntaub ntawv ntxiv. Tag nrho cov ion chromatogram nyob rau hauv qhov tsis zoo ion hom tau pom nyob rau hauv daim duab S1. MS cov ntaub ntawv tau txais kev tso cai los ntawm kev sib piv nrog cov ntaub ntawv hauv tsab ntawv tshaj tawm dhau los (Li et al., 2015) thiab sau tseg hauv Table S1. Cov txheej txheem tshuaj ntsuam xyuas kom muaj nuj nqis ntawm cov cim kuj tau raug lees paub thiab piav qhia luv luv hauv cov ntaub ntawv ntxiv. HPLC chromatograms tau nthuav tawm hauv daim duab 1. Peb qhov tseem ceeb (echinacoside, acteoside, thiab isoacteoside) ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa)tau txheeb xyuas piv nrog cov ntaub ntawv siv. Kev txheeb xyuas ntawm cov tebchaw thiab lawv cov concentration hauv TGCT tau qhia hauv Table 1.

3.2. Cov teebmeem ntawm TGCT ntawm BW thiab calorie kom tsawg

Raws li pom hauv daim duab 2A, nas noj HFSD pom qhov hnyav nce los ntawm qhov nruab nrab ntawm 41 g piv nrog NC pab pawg tom qab 4 lub lis piam. Txawm li cas los xij, STZ pom tseeb txo qis BW ntawm nas thaum piv nrog NC pab pawg. Hauv qhov sib piv, BW hauv TGCT pawg (100 thiab 200 mg / kg) tau nce zuj zus thiab nce ntxiv (p <0.05) los="" ntawm="" 8.1="" feem="" pua="" ​​​​thiab="" 11.1="" feem="" pua="" ​​​​ntawm="" qhov="" piv="" nrog="" dc="" pawg="" thaum="" kawg="" ntawm="" lub="" sijhawm="" sim="" ,="" uas="" qhia="" tias="">(tag nrho glycosides ntawmCistanchetubulosa)tuaj yeem tiv thaiv kev poob phaus ntau dhau hauv cov kab mob pathological. Cov calorie kom tsawg ntawm NC pab pawg tau pom tau qis dua li lwm pab pawg, thiab tsis muaj qhov sib txawv ntawm cov calorie kom tsawg ntawm lwm pawg tsib (Fig. 2B).

3.3. Cov teebmeem ntawm TGCT ntawm FPG, OGTT, thiab HbA1c

STZ-induced ntshav qab zib nas pom tau tias muaj kev nce ntxiv hauv FPG piv nrog NC pawg (p < 0.01)="" raws="" li="" tau="" piav="" qhia="" hauv="" daim="" duab="" 3a.="" kev="" tswj="" hwm="" qhov="" ncauj="" ntawm="">(tag nrho glycosides ntawmCistanchetubulosa)Ua kom pom cov nyhuv hypoglycemic nyob rau hauv ib lub sij hawm thiab koob tshuaj-nyob ntawm tus yam ntxwv. TGCT(tag nrho glycosides ntawmCistanchetubulosa)(100 thiab 200 mg/kg) txo qis FPG theem ntawm 4th (22.1 feem pua ​​thiab 24.8 feem pua) thiab 6th (23.2 feem pua ​​thiab 56.4 feem pua) lub lis piam piv nrog DC pawg. Raws li tau piav qhia hauv daim duab 3B thiab C, TGCT (100 mg / kg thiab 200 mg / kg) pom tseeb txo cov ntshav qabzib los ntawm 16.1 feem pua ​​​​thiab 22.2 feem pua ​​​​ntawm 0.5 h thiab txo los ntawm 17.2 feem pua ​​thiab 26.5 feem pua ​​​​ntawm 1 h, thiab AUC -G ntawm TGCT pawg kuj raug txo los ntawm 8.1 feem pua, 18.5 feem pua, thiab 25.4 feem pua. Raws li tau piav qhia hauv daim duab 3D, muaj qhov nce ntxiv hauv HbA1c (93.3 feem pua) piv nrog NC pawg, thaum kev tswj qhov ncauj ntawm TGCT (100 mg / kg thiab 200 mg / kg) rau cov nas mob ntshav qab zib zoo kawg li (p <0.05) txo="" qis.="" hba1c="" (26.7="" feem="" pua="" ​​thiab="" 37.4="" feem="" pua)="" piv="" nrog="" dc="">

3.4. Glycogen cov ntsiab lus hauv daim siab thiab cov leeg

Raws li pom hauv daim duab 4A thiab B, qib glycogen tau txo qis hauv cov nas ntshav qab zib. Thaum sib txawv ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa)tau muab tshuaj rau cov nas mob ntshav qab zib rau 6 lub lis piam, daim siab glycogen hauv pawg TGCT (100 mg / kg thiab 200 mg / kg) siab dua (25.2 feem pua ​​​​thiab 40.8 feem pua) dua li cov hauv pawg DC (p <0.05). ,="" fig.="" 4a).="" ib="" qho="" txiaj="" ntsig="" zoo="" sib="" xws="" tau="" pom="" hauv="" cov="" leeg="" nqaij="" uas="" glycogen="" hauv="">(tag nrho glycosides ntawmCistanchetubulosa)pawg (100 mg/kg thiab 200 mg/kg) siab dua (40.7 feem pua ​​​​thiab 52.6 feem pua ​​) dua li cov hauv pawg DC (p <0.05, Fig. 4B).

3.5. Cov teebmeem ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa)Hauv cov tshuaj insulin, C-Peptide, thiab ISI

Raws li qhia hauv Table 2, insulin thiab C-peptide tau pom tseeb (p < 0.01)="" txo="" qis="" hauv="" cov="" nas="" ntshav="" qab="" zib="" piv="" nrog="" nc="" pawg.="" txawm="" li="" cas="" los="" xij,="">(tag nrho glycosides ntawmCistanchetubulosa)at all doses caused no significant increase in insulin and C-peptide levels (p >{{0}}.05), txawm hais tias insulin thiab C-peptide hauv pawg TGCT-kho tau siab dua me ntsis hauv pawg DC. Lub caij no, peb suav ISI hauv daim duab 4C. Hauv kev sib piv rau cov tshuaj insulin secretion, ISI tau nce 32.9 feem pua ​​thiab 37.8 feem pua ​​​​los ntawm TGCT (100 mg / kg thiab 200 mg / kg) piv nrog DC pawg (p <>

3.6. Kev kuaj histopathological ntawm pancreas

Txhawm rau txheeb xyuas cov txiaj ntsig ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa)Hauv kev tsim cov kab mob pancreatic islets, kev soj ntsuam histological ntawm cov txiav tau ua tiav. Nyob rau hauv daim duab 5A, ib txwm histological qauv thiab loj islets tau pom nyob rau hauv pawg NC. Hauv qhov sib piv, STZ-txhaj tshuaj ua rau txo tus lej thiab txoj kab uas hla ntawm cov islets nrog cov cim hloov pauv microvesicular (Fig. 5B). Ob leeg TGCT(tag nrho glycosides ntawmCistanchetubulosa)thiab tau ntsib ntau ntxiv thiab qhov loj ntawm cov islets los ntawm kev txheeb xyuas cov qhab nia (Fig. 5C–F).

3.7. Cov teebmeem ntawm TGCT ntawm p-PKB, p-GSK3, HK thiab PK

Hauv Table 2, cov qib phosphorylation ntawm PKB thiab GSK3 tau pom tias muaj ntau heev (p < 0.05)="" txo="" qis="" hauv="" cov="" nas="" mob="" ntshav="" qab="" zib.="">(tag nrho glycosides ntawmCistanchetubulosa)(100 mg/kg thiab 200 mg/kg) zoo kawg nkaus ua kom cov concentrations p-PKB (TGCT-100: 13.5 feem pua ​​, p < 0="" .="" cov="" teebmeem="" zoo="" sib="" xws="" ntawm="" tgct="" ntawm="" hk="" thiab="" pk="" kuj="" tau="" pom.="" tgct="" pawg="" tau="" nce="" cov="" dej="" num="" ntawm="" hk="" (tgct{19}}:="" 30.2="" feem="" pua,="" p=""><0.05 thiab="" tgct-200:="" 59.1="" feem="" pua,="" p=""><0.01) thiab="" pk="" (tgct-200:="" 32.7="" feem="" pua,="" p=""><0.05) dua="" li="" cov="" hauv="" pawg="">

3.8. Cov teebmeem ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa)ntawm dyslipidemia

Raws li pom nyob rau hauv Table 3, Cov nas kho nrog TGCT (100 mg/kg thiab 200 mg/kg) yog qhov tshwj xeeb ameliorated lipid txawv txav. TG raug txo los ntawm 19.8 feem pua ​​(p <0.05) thiab="" 25.9="" feem="" pua="" ​​(p=""><0.01); tc="" raug="" txo="" los="" ntawm="" 28.5="" feem="" pua="" ​​thiab="" 31.4="" feem="" pua="" ​​(p=""><0.05); ldl-c="" raug="" txo="" los="" ntawm="" 20.0="" feem="" pua="">< 0.01)="" compared="" with="" dc="" group.="" however,="" the="" suppressed="" hdl-c="" level="" in="" dc="" group="" was="" significantly="" elevated="" by="" 26.8%="" (p="" <="" 0.05)="" in="" tgct-200="">

3.9. Cov teebmeem ntawm TGCT ntawm oxidative kev nyuaj siab thiab o

Nyob rau hauv lub teeb ntawm lub luag hauj lwm tseem ceeb ntawm oxidative kev nyuaj siab thiab inflamma tion nyob rau hauv lub pathophysiology ntawm ntshav qab zib, peb soj ntsuam lub peev xwm ntawm TGCT ntawm oxidative kev nyuaj siab thiab o nyob rau hauv cov nas mob ntshav qab zib. TGCT(tag nrho glycosides ntawmCistanchetubulosa)(100 thiab 200 mg/kg) zoo nce (14.7 feem pua, p <0.05 thiab 20.5 feem pua ​​, p< 0.01)="" the="" activity="" of="" sod,="" remarkably="" elevated="" (16.3%,="" p="" <="" 0.01="" and="" 22.3%,="" p="" <="" 0.01)="" the="" gsh-px="" activity="" and="" significantly="" decreased="" (15.0%,="" p="" <="" 0.05="" and="" 19.7%,="" p="" <="" 0.05)="" the="" mda="" formation="" compared="" with="" dc="" group="" in="" table="" 3.="" similarly,="" tgct="" (200="" mg/kg)="" treatment="" also="" blocked="" the="" stz-induced="" overproduction="" of="" pro-inflammatory="" cytokines="" tnf-α="" (21.8%,="" p="" <="" 0.01),="" il-6="" (14.0%,="" p="" <="" 0.05)="" and="" il-1β="" (15.2%,="" p=""><>

4. Kev sib tham

Mob ntshav qab zib mellitus yog ib qho mob hnyav thiab mob ntev uas tshwm sim los ntawm hyperglycemia. Tam sim no, FPG yog ib qho kev ntsuas tshwj xeeb ntawm cov ntshav qabzib ntau, OGTT yog qhov ntsuas ntsuas qhov ntsuas ntawm qhov txawv txav thaum ntxov ntawm cov piam thaj pov tseg, thaum HbA1c tau siv dav los ua tus qauv ntsuas kub rau kev tswj glycemic qhia txog qib qabzib nruab nrab ntawm 120 hnub ua ntej qhov kev xeem ( Nagy et al., 2018; Gan et al., 2018). FPG, OGTT, thiab HbA1c yog siv rau hauv kev kho mob rau kev kuaj mob thiab kev tswj cov kab mob prediabetes thiab ntshav qab zib (Chai et al., 2017). Hauv peb txoj kev tshawb fawb, nws pom tseeb tias FPG ntawm cov nas mob ntshav qab zib tau kho nrog TGCT(tag nrho glycosides ntawmCistanchetubulosa)(100 mg / kg thiab 200 mg / kg) tau poob qis dua piv nrog DC pawg. Raws li cov ntaub ntawv qhia nyob rau hauv OGTT thiab AUC-G, cov piam thaj tsis txaus siab thiab cov piam thaj nce siab tau thim rov qab los ntawm TGCT hauv cov nas mob ntshav qab zib. Lub caij no, qhov tshwm sim kuj tau txais kev txhawb nqa los ntawm kev ntsuas HbA1c cov ntsiab lus. Cov ntaub ntawv no qhia tias TGCT(tag nrho glycosides ntawmCistanchetubulosa)ameliorated lub physiological indexes ntawm cov kab mob ntshav qab zib nas los ntawm kev tswj cov ntshav qabzib homeostasis.

Cov tshuaj insulin tsis txaus thiab IR ua lub luag haujlwm tseem ceeb hauv kev txhim kho hyperglycemia. Targeting ob leeg yog tsim nyog los txhim kho glycemic tswj thiab tiv thaiv T2DM (Szoke thiab Gerich, 2005; Punthakee li al., 2018). Ntau tus kws tshawb fawb tau tshaj tawm tias kev sib xyaw ua ke ntawm kev noj zaub mov muaj roj ntau thiab cov tshuaj STZ qis yog ib qho txiaj ntsig zoo ntawm kev ua rau T2DM hauv kev sim nrog tsiaj. Cov koob tshuaj STZ qis ua rau muaj kev cuam tshuam me me ntawm insulin secretion uas zoo sib xws rau cov theem tom qab ntawm T2DM (Gheibi li al., 2017). Hauv txoj kev tshawb no, TGCT tsis muaj qhov nce ntxiv ntawm cov tshuaj insulin los yog rov qab kho cov kab mob pancreas ntawm cov nas mob ntshav qab zib, txawm tias cov tshuaj insulin thiab cov islets hauv TGCT-kho cov pab pawg tau siab dua li ntawm pawg DC. Cov kev tshawb pom no yog ua raws li txoj kev tshawb fawb yav dhau los hauv db/db nas (Xiong li al., 2013). Peb cov txiaj ntsig tau pom qhov hnyav nce ntxiv hauv cov nas ntshav qab zib kho nrog TGCT(tag nrho glycosides ntawmCistanchetubulosa), uas tuaj yeem piav qhia los ntawm kev nce me ntsis ntawm insulin uas tuaj yeem cuam tshuam cov protein catabolism hauv cov leeg nqaij (Adams li al., 2019). Tsis tas li ntawd, qhov tshwm sim ntawm ISI tau nthuav tawm tias TGCT pom tseeb tau kho IR ntawm cov nas ntshav qab zib uas zoo ib yam nrog cov ntawv tshaj tawm dhau los (Kong et al., 2018), uas tau muab cov pov thawj tshiab hais txog lub peev xwm ntawm cov tshuaj tiv thaiv kab mob ntawm TGCT.

Raws li peb txhua tus paub, PKB / GSK3 txoj hauv kev yog ib txoj hauv kev tseem ceeb tshaj plaws ntawm kev qhia cov tshuaj insulin, tau raug qhia los kho cov tshuaj insulin-induced glycogen synthesis (Zheng li al., 2015). HK thiab PK ua raws li lub hom phiaj tshuaj muaj peev xwm hauv kev kho tshuaj kho mob ntshav qab zib. Kev ua haujlwm qis HK thiab PK tau lees paub hauv IR, thaum ua kom HK thiab PK ua rau muaj glycogen reserves ntau dua lossis glycolysis ua kom muaj zog txaus los ntawm kev siv cov ntshav qabzib (Hu et al., 2014). Hauv kev tshawb fawb tam sim no, TGCT(tag nrho glycosides ntawmCistanchetubulosa)Kev kho mob ib txhij nce cov phosphorylated proteins qhia ntawm PKB thiab GSK3, ua rau muaj kev cuam tshuam tseem ceeb hauv cov dej num ntawm HK thiab PK, thiab rov qab kho cov ntsiab lus glycogen hauv daim siab thiab cov leeg vim cov ntshav qabzib txo qis. Cov txiaj ntsig no qhia tias TGCT(tag nrho glycosides ntawmCistanchetubulosa)Ua kom cov enzymes tseem ceeb ntawm txoj kev taw qhia cov tshuaj insulin, thiab ntxiv cov pov thawj tias cov tshuaj insulin rhiab heev hauv cov nas mob ntshav qab zib.

Kev mob ntshav qab zib ntev ntev kuj tseem txhawb nqa LDL-C thiab txo qis HDL-C qib uas ua rau lipid dysregulation (Jayashankar et al., 2016), thiab dyslipidemia yog ib qho cim tsim rau endothelial dysfunction thiab cardiovascular kev pheej hmoo ntawm ntshav qab zib (Shahwan li al., 2019). Hauv peb txoj kev tshawb fawb, TGCT (200 mg / kg) zoo kawg nkaus txo qis ntawm TC, TG, thiab LDL-C, thiab txhim kho qib HDL-C hauv cov nas mob ntshav qab zib, uas zoo ib yam nrog cov ntaub ntawv dhau los.Cistanchetubulosatswj cov lipid metabolism hauv nas (Shimoda li al., 2009; Xiong et al., 2013). Cov kev tshawb pom no qhia tias TGCT(tag nrho glycosides ntawmCistanchetubulosa)Tej zaum yuav muaj txiaj ntsig zoo rau cov neeg mob ntshav qab zib uas muaj ntshav lipid txawv txav.

STZ yog ib hom tshuaj tua kab mob dav dav, uas muaj kev xaiv toxicity ntawm cov kab mob pancreatic -cells uas tshwm sim los ntawm kev nce ntawm superoxide radical, thiab ntxiv rau kev tswj tsis zoo glycemic nyob rau hauv lem (Ghosh li al., 2015; Swain et al., 2020 ). Lub caij no, oxidative kev nyuaj siab kuj yog ib qho tseem ceeb ua rau IR nyob rau hauv ntau qhov chaw (Taniguchi li al., 2006). IR thiab ntshav qab zib muaj feem cuam tshuam nrog kev txo qis ntawm cov enzymes antioxidant, xws li SOD thiab GSH-Px (Styskal et al., 2012). Cov kev tshawb fawb yav dhau los tau hais tias kev txo qis oxidative kev nyuaj siab tuaj yeem txo cov ntshav qabzib hauv cov nas ntshav qab zib (Lim li al., 2012; Gao et al., 2016). Hauv txoj kev tshawb no, kev kho mob nrog TGCT(tag nrho glycosides ntawmCistanchetubulosa)rov qab kho cov cellular tiv thaiv kev ua haujlwm ntawm SOD thiab GSH-Px thiab txo MDA qib hauv cov nas mob ntshav qab zib, uas pom tau tias TGCT muaj cov khoom hauv kev tiv thaiv oxidation.

Cov ntsiab lus tseem ceeb hauv TGCT(tag nrho glycosides ntawmCistanchetubulosa)yog PhGs thiab tag nrho cov ntsiab lus ntawm echinacoside, acteoside thiab isoacteoside yog ntau tshaj 661 mg / g. Cov nroj tsuag uas muaj cov tshuaj tiv thaiv antioxidant siab, xws li PhGs, feem ntau tau ua pov thawj tias muaj cov nyhuv hypoglycemic (Morikawa li al., 2014; Shimada li al., 2017; Spínola li al., 2019). Yog li, peb kwv yees tias cov tshuaj tiv thaiv kab mob ntshav qab zib ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa)tej zaum yuav raug ntaus nqi los ntawm kev tiv thaiv oxidant ntawm PhGs. Hauv kev xav txog cov ntsiab lus ntawm echinacoside hauv TGCT, cov kev tshawb fawb ntxiv yuav tsum tau tshawb xyuas cov teebmeem ntawm echinacoside ntawm IR thiab ntshav qab zib hauv vivo thiab hauv vitro qauv.

Nws yog dav paub tias mob ntshav qab zib mellitus yog ib yam kab mob inflammatory. Inflammatory cytokines xws li TNF-, IL-6, thiab IL-1 tuaj yeem cuam tshuam nrog cov insulin receptor signaling txoj kev, thiab ua rau IR (Bastard li al., 2006). Kev mob tshwm sim zoo li yog lub hom phiaj siv tshuaj hauv kev kho mob IR, thiab los ntawm kev mob ntshav qab zib ntxiv (Chen li al., 2015). Hauv kev tshawb fawb tam sim no, TGCT(tag nrho glycosides ntawmCistanchetubulosa)tuaj yeem txo qib ntawm TNF- , IL-6, thiab IL-1 , uas qhia tau tias muaj kev tiv thaiv kab mob. Qhov no yuav yog lwm txoj kev tiv thaiv kab mob ntshav qab zib ntawm TGCT hauv cov nas mob ntshav qab zib.

Cov ntaub ntawv pov thawj qhia tau tias cov ntshav qabzib siab tsis tu ncua ua rau muaj qhov txawv txav hauv cov qauv thiab kev ua haujlwm ntawm cov protein thiab lipids, uas ua rau glycoxidation thiab peroxidation, thiab tom qab ntawd txhawb kev tsim cov cytokines. Ib yam li ntawd, nce inflammatory cytokines ua rau zus tau tej cov reactive oxygen hom thiab lwm yam reactive moieties, uas txhawb oxidative kev nyuaj siab thiab oxidative puas. Qhov no tshwm sim nyob rau hauv ib tug vicious voj voog (Aghadavod li al., 2016; Domingueti li al., 2016). Yog li, txo cov ntshav qabzib thiab lipids tuaj yeem ua rau muaj kev tiv thaiv antioxidant thiab tiv thaiv kev mob ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa). Rau qee qhov, qhov tseeb antidiabetic mechanism tsis meej meej thiab xav tau kev tshawb nrhiav ntxiv.

5. Cov lus xaus

Hauv kev xaus, qhov kev tshawb fawb no qhia tias TGCT(tag nrho glycosides ntawmCistanchetubulosa)yog tus neeg sawv cev zoo rau kev kho mob hyperglycemia thiab hyperlipidemia hauv kev noj haus / STZ-vim cov nas mob ntshav qab zib. Tsis tas li ntawd, cov nyhuv antidiabetic tej zaum yuav muaj feem cuam tshuam nrog cov tshuaj tua kab mob antioxidant thiab tiv thaiv kab mob ntawm TGCT. Txoj kev tshawb no yuav nthuav tawm qhov ua tau los qhia TGCT(tag nrho glycosides ntawmCistanchetubulosa)hauv kev kho mob ntshav qab zib. Txawm li cas los xij, cov ncauj lus kom ntxaws antidiabetic mechanism ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa)tseem tsis paub meej, thiab kev tshawb fawb ntxiv hauv vivo thiab hauv vitro yog qhov tsim nyog.

Sau cov nyiaj pab

Lub tswv yim thiab tsim: ZQM, KNZ, thiab WX. Kev npaj ntawm TGCT(tag nrho glycosides ntawmCistanchetubulosa), chemical constituents analysis, and methodology validation: RG, ZKX, and WJL. Mob ntshav qab zib induction thiab kev kho mob txhua hnub: KNZ thiab HF. Kev npaj cov qauv rau kev soj ntsuam biochemical: KNZ, HF, thiab ZQM. Kev txheeb xyuas cov ntaub ntawv thiab sau cov ntawv sau: KNZ, ZQM, thiab YST. Cov ntaub ntawv validation thiab phau ntawv hloov kho: WZH, GD, thiab YST.

Tshaj tawm ntawm kev sib tw txaus siab

Cov kws sau ntawv tau tshaj tawm tias tsis muaj kev cuam tshuam ntawm kev txaus siab.

Kev lees paub

Txoj kev tshawb no tau txais kev txhawb nqa nyiaj txiag los ntawm National Science & Technology Major Project Key New Drug Creation and Manufacturing Program, Tuam Tshoj (Grant no. 2013ZX09402203), thiab Science thiab Technology Project ntawm Huzhou, Zhejiang Province (Grant no. 2018GZ03). Peb ua tsaug rau Mrs. Ning Su thiab Mrs. Sha Liu los ntawm Jiangsu lub koom haum tshuaj rau lawv cov kev pab hauv cov ntaub so ntswg. Jinfeng Wu thiab Xuefei Zhang thiab lwm yam tau txais txiaj ntsig zoo hauv qhov kev tshawb fawb no.

Daim Ntawv Ntxiv A. Cov ntaub ntawv ntxiv rau tsab xov xwm no tuaj yeem pom hauv online ntawm https://doi. org/10.1016/j.jep.2021.113991.

Los ntawm: 'Hypoglycemic thiab hypolipidemic cuam tshuam ntawm tag nrho glycosides ntawmCistanchetubulosahauv kev noj haus / streptozotocin-induced mob ntshav qab zib nas 'los ntawmKuiniu Zhu, et al

---Journal of Ethnopharmacology 276 (2021) 113991