Hypoglycemic Thiab Hypolipidemic Los Ntawm Tag Nrho Glycosides Ntawm Cistanche Tubulosa
Mar 28, 2022
Hu rau:joanna.jia@wecistanche.com/ WhatsApp: 008618081934791
Kuiniu Zhu a,b,1, Zhaoqing Meng a,c,1, Yushan Tian d, Rui Gu a, Zhongkun Xu a, Hui Fang a, Wenjun Liu a, Wenzhe Huang a, Gang Ding a, Wei Xiao a,*
TSAB NTAWV
Ethnopharmacological qhov tseeb:Cistanche tubulosa (Schrenk) R. Wight (Orobanchaceae) yog ib qho tshuaj nquag siv hauv ntau cov tshuaj ntsuab tshuaj uas siv los kho ntshav qab zib hauv Suav teb. Hauv cov kev tshawb fawb tsis ntev los no, cov tshuaj tiv thaiv kab mob ntawm Cistanche tubulosa extracts tau lees paub. Txawm li cas los xij, tsis muaj kev tshawb fawb tau qhia txog tag nrho glycosides ntawm Cistatnche tubulosa (TGCT).
Lub hom phiaj ntawm kev kawm:Txoj kev tshawb fawb tam sim no tsom los tshawb xyuas cov teebmeem hypoglycemic thiab hypolipidemic ntawm TGCT thiab cov txheej txheem muaj peev xwm hauv kev noj zaub mov / streptozotocin (STZ)-induced ntshav qab zib nas, thiab tshuaj lom neeg cov ntsiab lus tseem ceeb ntawm TGCT.
Cov ntaub ntawv thiab cov txheej txheem:Cov ntsiab lus tseem ceeb ntawm TGCT yog tus cwj pwm los ntawm HPLC/Q-TOF-MS thiab qhov 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 qabzib ntsuas (OGTT), thaj tsam hauv qab nkhaus ntawm qabzib (AUC-G), glycosylated hemoglobin (HbA1c), yoo mov insulin, ntshav C-peptide, glycogen cov ntsiab lus, thiab cov tshuaj insulin rhiab heev. . 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 Kev tshuaj ntsuam xyuas tshuaj tau qhia 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 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 ntawm inhibitions oxidative stress thiab o.
Cistanche attributionrau cov dej num ntawm inhibitions ntawmoxidative kev nyuaj siabthiabmob.
1. Taw qhia
Mob ntshav qab zib mellitus yog suav tias yog kab mob metabolic, uas yog tus cwj pwm los ntawm hyperglycemia 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 insulin) muaj txiaj ntsig zoo hauv kev 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).
Cistanche tubulosa (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 tsis 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 cov tshuaj aqueous extract ntawm Cistanche tubulosa tau pom 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 ntshav qab zib nas (Kong et al., 2018). Phenylethanoid glycosides (PhGs) yog cov khoom tseem ceeb ntawm Cistanche tubulosa (Morikawa li al., 2014), uas tau nthuav tawm ntau yam kev ua ub no xws li tshuaj tiv thaiv oxidation (Xue et al., 2017) thiab anti-cancer (Fu et al., 2019) . Tsis tas li ntawd, PhGs cuam tshuam qhov nce ntawm cov ntshav qabzib hauv cov ntshav qabzib hauv cov hmoov txhuv nplej siab (Morikawa li al., 2014), inhibited sodium-dependent glucose 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 ntawm Cistanche tubulosa (TGCT).
Hauv txoj kev tshawb fawb tam sim no, cov tshuaj tiv thaiv kab mob ntawm TGCT tau raug soj ntsuam hauv kev noj zaub mov muaj roj siab-sucrose (HFSD) thiab STZ-vim cov nas mob ntshav qab zib. 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.
cistanche txiaj ntsig extract hmoov
2.2. Cov peev txheej cog thiab kev npaj ntawm TGCT
Qhuav succulent qia ntawm Cistanche tubulosa tau 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. Ib daim ntawv pov thawj (No. 20161103) raug crushed rau hauv cov hmoov thiab muab tso rau hauv lub herbarium ntawm Jiangsu Kanion Pharmaceutical Co., Ltd. Raws li SFDA lub teb chaws tshuaj standard "Congrong Zonggan Jiaonang " (YBZ07482005-2011Z), cov tshuaj crude (1 kg) tau muab rho tawm nrog dej 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 (0.21 mg) thiab peb cov qauv sib xyaw (echinacoside: 131.3 ug, acteoside: 4.2 ug, isoacteoside: 39.4 ug) tau sib cais 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; tawg voltage, 135V; 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. Kev txheeb xyuas ntau ntawm TGCT 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 (CMCNa, 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.
Cistanche tiv thaiv daim siab.
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.
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 60 μL cov ntshav kuaj tau sau nrog lub raj xa dej los ntawm lub qhov txhab orbital (0 h), tom qab ntawd muab tshuaj nrog TGCT (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. Lub cheeb tsam nyob rau hauv qhov nkhaus ntawm qabzib (AUC-G) raug xam 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 tau excised, yaug, thiab tsau nyob rau hauv 10 feem pua ntawm cov formalin, ces dehydrated nyob rau 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 haematoxylin-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
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 tau txheeb xyuas piv nrog cov khoom 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="" tau="" qhia="" tias="" tgct="" tuaj="" yeem="" tiv="" thaiv="" kev="" poob="" phaus="" ntau="" dhau="" los="" ntawm="" 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.="">
3.3. Cov teebmeem ntawm TGCT ntawm FPG, OGTT thiab HbA1c
STZ-induced mob ntshav qab zib nas pom tau tias muaj kev nce ntxiv hauv FPG piv nrog NC pab pawg (p < 0.01)="" raws="" li="" tau="" piav="" qhia="" hauv="" daim="" duab="" 3a.="" kev="" tswj="" qhov="" ncauj="" ntawm="" tgct="" pom="" tau="" tias="" muaj="" cov="" nyhuv="" hypoglycemic="" nyob="" rau="" hauv="" ib="" lub="" sij="" hawm="" thiab="" koob="" tshuaj-nyob="" ntawm="" seb.="" tgct="" (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="" tau="" 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="">
Fig. 1. HPLC spectrum ntawm TGCT thiab cov qauv tshuaj ntawm cov tebchaw loj hauv TGCT.
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 qhov sib txawv ntawm TGCT tau muab rau cov nas ntshav qab zib rau 6 lub lis piam, daim siab glycogen hauv TGCT pawg (100} mg / kg thiab 200 mg / kg) siab dua (25.2 feem pua thiab 40.8 feem pua) dua. cov hauv pawg DC (p <0.05, fig.="" 4a).="" cov="" txiaj="" ntsig="" zoo="" sib="" xws="" tau="" pom="" hauv="" cov="" leeg="" uas="" glycogen="" hauv="" tgct="" pawg="" (100="" mg="" kg="" thiab="" 200="" mg="" kg)="" siab="" dua="" (40.7="" feem="" pua="" thiab="" 52.6="" feem="" pua)="" dua="" li="" cov="" hauv="" dc="" pawg="" (p=""><0.05, fig.="">
3.5. Cov teebmeem ntawm TGCT ntawm cov tshuaj insulin, C-Peptide thiab ISI
As shown in Table 2, insulin and C-peptide were obviously (p < 0.01) decreased in diabetic rats compared with the NC group. However, TGCT 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 <>
Table 1 Cov ntsiab lus ntawm phytochemical markers ntawm TGCT.
3.6. Kev kuaj histopathological ntawm pancreas
Txhawm rau txhawm rau txheeb xyuas qhov cuam tshuam ntawm TGCT ntawm kev rov tsim dua tshiab hauv pancreatic islets, histological tsom xam ntawm txiav txiav tau ua. 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 thiab Met tau nce tus lej 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 yog qhov tseem ceeb (p < 0.{{10}}}5)="" txo="" qis="" hauv="" cov="" nas="" mob="" ntshav="" qab="" zib.="" tgct="" (100="" mg/kg="" thiab="" 200="" mg/kg)="" zoo="" kawg="" nkaus="" ua="" kom="" cov="" concentrations="" p-pkb="" (tgct-100):="" 13.5="" feem="" pua="" ,="" p="">< 0.05="" thiab="" tgct-200:="" 16.7="" feem="" pua="" ,="" p="">< 0.05)="" thiab="" p="" gsk3="" (tgct-200:="" 18.3="" feem="" pua="" ,="" p="">< 0.01).="" cov="" teebmeem="" zoo="" sib="" xws="" ntawm="" tgct="" ntawm="" hk="" thiab="" pk="" kuj="" tau="" pom.="" tgct="" pawg="" tau="" nce="" cov="" dej="" num="" ntawm="" hk="" (tgct-100:="" 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="">
Daim duab 2. Cov teebmeem ntawm TGCT ntawm lub cev qhov hnyav (A) thiab calorie kom tsawg (B) hauv STZ-induced ntshav qab zib nas.
3.8. Cov teebmeem ntawm TGCT 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 o 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 (100 thiab 200 mg/kg) tau nce zoo (14.7 feem pua, p <0.05 thiab="" 20.5="" feem="" pua="" ,="">< 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 (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 tau qhia tias TGCT ameliorated physiological indexes ntawm cov nas ntshav qab zib los ntawm kev tswj cov ntshav qabzib homeostasis.
Fig. 3. Kev hloov pauv ntawm FPG hauv cov nas mob ntshav qab zib thaum raug tswj hwm nrog TGCT rau 6 lub lis piam.
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 hom kev tshawb fawb tau tshaj tawm tias kev sib xyaw ua ke ntawm kev noj zaub mov muaj roj ntau thiab cov tshuaj STZ tsawg 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 ua rau muaj qhov nce ntxiv ntawm cov tshuaj insulin los yog rov ua kom 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 mob ntshav qab zib tau kho nrog TGCT, uas tuaj yeem piav qhia los ntawm kev nce me ntsis ntawm cov tshuaj 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 qhov muaj 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). Nyob rau hauv txoj kev tshawb no, TGCT kev kho mob ib txhij nce cov phosphorylated proteins qhia ntawm PKB thiab GSK3, coj mus rau ib tug tseem ceeb reversal nyob rau hauv cov dej num ntawm HK thiab PK, thiab markedly rov qab glycogen ntsiab lus nyob rau hauv lub siab thiab cov leeg raws li cov ntshav qabzib txo. Cov txiaj ntsig no tau qhia tias TGCT tau qhib 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.
Table 2 Cov teebmeem ntawm TGCT ntawm insulin secretion thiab carbohydrate metabolizing enzymes hauv cov nas 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 li al., 2016), thiab dyslipidemia yog ib qho cim tsim rau endothelial dysfunction thiab plawv plawv mob 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 uas Cistanche tubulosa tswj cov lipid zoo. metabolism hauv nas (Shimoda et al., 2009; Xiong et al., 2013). Cov kev tshawb pom no tau qhia tias TGCT yuav muaj txiaj ntsig zoo dua rau tus 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 tau 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 yog PhGs thiab tag nrho cov ntsiab lus ntawm echinacoside, acteoside thiab isoacteoside yog ntau dua 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 ntawd, peb xav tias cov tshuaj tiv thaiv mob ntshav qab zib mellitus ntawm TGCT tej zaum yuav yog ib feem ntawm cov tshuaj antioxidant 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 txoj kev tshawb fawb tam sim no, TGCT tuaj yeem txo qib ntawm TNF- , IL-6, thiab IL-1 , uas qhia tau hais tiasanti-inflammatory nyhuv. 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. Rau qee qhov, qhov tseeb antidiabetic mechanism tsis meej meej thiab xav tau kev tshawb nrhiav ntxiv.
5. Cov lus xaus
Hauv cov ntsiab lus, txoj kev tshawb no tau qhia tias TGCT 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 hauv kev kho mob ntshav qab zib. Txawm li cas los xij, cov ncauj lus kom ntxaws antidiabetic mechanism ntawm TGCT tseem tsis paub meej, thiab kev tshawb fawb ntxiv hauv vivo thiab hauv vitro yog qhov tsim nyog.
Cistanche yog tus neeg sawv cev zoo rau kev kho mobhyperglycemiathiabhyperlipidemia.
