Empagliflozin Ameliorates Cov tsos mob ntawm Mob Ntshav Qab Zib Thiab Lub raum Tubular Dysfunction nyob rau hauv ib tus nas Model ntawm Diabetes Nrog raum loj (DEK)
Mar 20, 2022
Hu rau: Audrey Hu Whatsapp / hp: 0086 13880143964 Email:audrey.hu@wecistanche.com
Ayaka Domon1, Kentaro KatayamaID1, Touko Sato2, Yuki Tochigi1, Hiroyuki Tazaki2, Hiroetsu SuzukiID1*
Abstract
Tom qab: Sodium-glucose cotransporter 2 (SGLT2) inhibitors tau dav siv los txo qis hyperglycemia. Txoj kev tshawb fawb tam sim no tau tshawb xyuas qhov cuam tshuam ntawm SGLT2 inhibitor, empagliflozin, ntawm hyperglycemia hauv cov qauv nas tshiab ntawm cov ntshav qab zib hom 2 uas tsis yog rog nrog loj.lub raum(DEK). Methods: Male DEK rats with non-fasting blood glucose concentrations ≤300 mg/dl and >300 mg / dl tau muab faib ua cov tsis muaj ntshav qab zib thiab ntshav qab zib, feem. Cov pab pawg ntawm cov neeg tsis muaj ntshav qab zib (tswj) thiab ntshav qab zib (DM-cont) nas tau noj cov tshuaj chow rau 12 lub lis piam, thaum lwm pab pawg neeg mob ntshav qab zib (DM-empa) nas tau noj cov kab mob uas muaj empagliflozin (300 mg / kg / hnub) rau 12 lub lis piam. . Cov ntshav qabzib, lub cev hnyav, qabzib kam rau siab, noj zaub mov thiab dej kom tsawg, tso zis ntim, ntshav plasma, thiab cov zis biochemical ntsuas, thiab cov pob txha pob txha pob txha, thiab lawvlub raumthiab pancreas tau soj ntsuam histologically. Cov txiaj ntsig: Kev kho mob nrog empagliflozin txo cov ntshav qabzib concentration thiab noj zaub mov hauv cov nas ntshav qab zib, tab sis inhibited poob ntawm adeps reins thiab ua rau lub cev hnyav. Empagliflozin attenuated polyuria thiab polydipsia tab sis nce plasma concentrations ntawm tag nrho cov cholesterol, sodium, thiab tag nrho cov protein mus rau ib tug ib txwm muaj. Empagliflozin kuj tseem txo cov zis tso zis ntawm cov protein thiab electrolytes thiab rov kho cov pob txha pob txha pob txha thiab cov ntshav plasma concentrations ntawm valine thiab isoleucine mus rau qib qub. Ntxiv mus, dilation ntawm lub raum tubules thiabraumkev loj hlob tsis tau attenuated nyob rau hauv pawg DM-empa. Xaus: Cov lus teb ntawm DEK nas rau empagliflozin txawv ntawm lwm cov tsiaj mob ntshav qab zib hom, qhia tias DEK nas muaj cov yam ntxwv tshwj xeeb rau kev kawm thiab tshuaj xyuas ntau yam teebmeem ntawm SGLT2 inhibitors. Cov kev tshawb pom no kuj tau qhia tias empagliflozin tuaj yeem ua kom cov metabolism hauv lub cev thiab txhim kholub raumtubule ua haujlwm hauvmob ntshav qab zibtej yam kev mob.
Cistanche tubulosa tiv thaivraumkab mob, nyem qhov no kom tau txais cov qauv
Taw qhia
Kev nthuav dav thoob ntiaj teb ntawm ntshav qab zib mellitus (DM) tseem nce ntxiv, nrog cov neeg mob kwv yees txog 700 lab los ntawm 2045 [1]. Kwv yees li ntawm 90-95 feem pua ntawm cov neeg mob no muaj hom 2 DM, uas yog tus cwj pwm los ntawm kev txo qis ntawm insulin tso tawm nrog lossis tsis muaj cov tshuaj insulin tsis kam [2] thiab yog ib qho kev pheej hmoo loj rau cov kab mob raum thiab cov hlab plawv [3]. Hom 2 DM yog qhov ua rau lub raum tsis ua haujlwm kawg (ESRD), nrog rau mob ntshav qab zib nephropathy, tseem hu ua ntshav qab zib.raumkab mob, ua lub luag haujlwm rau ESRD hauv 20-40 feem pua ntawm cov neeg mob no [4]. Cov neeg mob ESRD yuav tsum tau lim ntshav lossis hloov lub raum kom tswj tau qhov qublub raum ua haujlwm. Cov txheej txheem no ua rau muaj kev cuam tshuam txog kev noj qab haus huv thiab nyiaj txiag rau tib neeg thiab tib neeg, nrog rau kev cuam tshuam rau tus neeg mob lub neej zoo [5]. Yog li kev siv zog yuav tsum tau los tiv thaiv lossis kho DM thiab mob ntshav qab zib nephropathy [6].
Sodium-glucose cotransporter 2 (SGLT2) inhibitors yog cov chav kawm tshiab ntawm cov tshuaj tiv thaiv kab mob ntshav qab zib uas ua rau nws tus kheej ntawm endogenous insulin [7]. SGLT2 inhibitors txo cov ntshav qabzib ntau los ntawm inhibiting qabzib reabsorption ntawm proximal raum tubules [7]. SGLT2 inhibitors kuj muaj txiaj ntsig zoo rau lub raum thiab cov hlab plawv cov teeb meem ntawm DM [8-10], nrog rau kev txhim kho cov metabolism [11], tag nrho cov no yog ib feem ntawm lawv cov kev txo qis hauv qabzib. Tsis tas li ntawd, SGLT2 inhibitors txhim kho lub raum cov txiaj ntsig hauv cov neeg mob uas tsis yog DM uas muaj mob ntevraumkab mob [12]. Txawm li cas los xij, cov teebmeem ntawm SGLT2 inhibitors, nrog rau cov txiaj ntsig ntawm lub raum, sib txawv ntawm cov neeg mob [13]. Cov kev hloov pauv no yuav cuam tshuam qhov sib txawv ntawm ntau yam, suav nrog cov caj ces predisposition, metabolic xwm txheej, hemodynamics, thiab kev ua neej.
Peb nyuam qhuav tsim ib qho tshiab nas hom kab mob uas tsis yog rog rog hom 2 DM nrog lojraum(DEK) los ntawm LEA. PET-tus tsiaj congenic strain. Hauv DEK hom, kwv yees li 50 feem pua ntawm cov txiv neej pom ntshav qab zib los ntawm 30 lub lis piam ntawm hnub nyoog. Cov nas mob ntshav qab zib no tau pom tias muaj kev txo qis hauv cov ntshav plasma insulin ntau ntxiv vim poob ntawm cov hlwb pancreatic, nce lub raum parenchyma, thiab ua kom pom qhov loj ntawm lub raum.lub raumnrog rau lub raum tubules [14] DEK nas kuj tau pom tias txo qis hauv cov ntshav plasma ntawm tag nrho cov cholesterol (TCHO), tag nrho cov protein, thiab albumin [14]. Cov kab mob histological thiab metabolic no tshwj xeeb rau DEK nas, qhov txawv ntawm cov qauv no los ntawm lwm cov tsiaj cov qauv ntawm cov ntshav qab zib thiab qhia tias DEK nas tuaj yeem pab tau rau kev kawm cov tshuaj pharmacological ntawm cov tshuaj tiv thaiv mob ntshav qab zib.lub raumthiab metabolism. Txoj kev tshawb no tam sim no, yog li, ntsuas qhov cuam tshuam ntawm SGLT2 inhibitor, empagliflozin ntawm lub raum thiab cov metabolism hauv cov kab mob DEK nas.
Materials and methods Animals Blood samples were obtained from the tail veins of non-fasting male DEK rats (total n = 33), and glucose concentrations were measured with GLUCOCARD PlushCare (Arkray Inc., Kyoto, Japan). Male DEK rats with non-fasting blood glucose concentrations≤300 mg/dl and >300 mg / dl tau muab faib ua nondiabetic thiab ntshav qab zib, raws li [14]. Tag nrho cov nas tau khaws cia nyob rau hauv cov xwm txheej, suav nrog lub teeb 14: 10 h: lub voj voog tsaus, chav tsev kub ntawm 20 ± 2˚C, thiab cov av noo ntawm 50 ± 10 feem pua. Txhua qhov kev tshawb fawb tsiaj tau pom zoo los ntawm Pawg Saib Xyuas Tsiaj thiab Siv Tsiaj ntawm Nippon Veterinary thiab Life Science University thiab tau ua raws li Cov Lus Qhia ntawm Tsiaj Saib Xyuas thiab Siv Pawg ntawm NipponVeterinary thiab Life Science University.
Male rats with blood glucose concentrations >300 mg / dl thaum muaj hnub nyoog 15-20 lub lis piam hauv DEK strain tau muab faib ua pawg empagliflozin noj (DM-empa) (n=5) thiab pawg zaub mov txheem (DM-cont) (n {{6}) }) hauv 2-3 lub lis piam tom qab tsim DM. Cov txiv neej nas uas tsis yog DM (cov ntshav qabzib tseem qis dua 300 mg / dl thaum muaj hnub nyoog 30 lub lis piam) hauv DEK hom tau txhais tias yog kev tswj hwm (n=12). Cov pab pawg tswj hwm thiab DM-cont muaj ad libitum nkag mus rau cov dej thiab cov khoom noj khoom haus (CR-LPF, Oriental Yeast Co., Ltd., Tokyo. Nyiv) rau 12 lub lis piam, thaum cov nas hauv pawg DM-empa muaj ad libitum nkag mus rau hauv dej. thiab kev noj zaub mov zoo ntxiv rau empagliflozin (300 mg / kg; Eli Lilly Nyiv; Kobe, Nyiv). Qhov koob tshuaj ntawm empagliflozin tau tsim los nrog kev siv rau cov ntaub ntawv dhau los [15, 16]. Cov ntshav qabzib tsis yoo mov, qhov hnyav ntawm lub cev, thiab kev noj zaub mov tau raug ntsuas txhua lub lim tiam. Txhua tus nas tau txi tom qab 12 lub lis piam, thiab lawvraum, pancreases, thiab cov ntshav kuaj tau sau tseg. Peb teeb tsa tib neeg qhov kawg rau txhua tus tsiaj uas txo 25 feem pua ntawm lub cev hnyav lossis pom cov tsos mob cuam tshuam nrog kev mob hnyav. Txawm li cas los xij, tsis muaj nas pom qhov txo qis ntawm lub cev hnyav thiab mob hnyav hauv txoj kev tshawb no.
Qhov ncauj Glucose Tolerance Test (OGTT)
Kev ntsuas qhov ncauj ntawm qhov ncauj (OGTT) tau ua hnub tom qab 12 lub lis piam kho mob, raws li tau piav qhia [14]. Luv luv, cov nas tau yoo mov rau 16 teev thiab muab 2g / kg lub cev qhov hnyav (BW) ntawm cov piam thaj hauv qhov ncauj. Cov ntshav tau txais los ntawm tus Tsov tus tw leeg, thiab ntsuas cov piam thaj hauv cov ntshav tau ntsuas 0, 30, 60, 120, 180, thiab 240 min tom qab kev thauj khoom qabzib. Cov ntshav kuaj kuj tau sau los ntawm cov hlab ntsha jugular nyob rau hauv isoflurane anesthesia tswj los ntawm cov tshuaj loog me me (TK-7, Biomachinery, Chiba, Nyiv) ntawm 0, 30, thiab 60 min tom qab cov piam thaj load, tov nrog heparin thiab centrifuged. rau 15 feeb ntawm 4˚C kom tau plasma. Kev ntsuas ntshav plasma insulin ntau npaum li cas tau ntsuas los ntawm LBIS Rat Insulin ELISA Kit (RTU) (Shibayagi, Gunma, Nyiv).
Kev ntsuas ntawm lub raum tsis ua haujlwm
Thaum lub asthiv 12, cov nas tau nyob ib leeg hauv cov kab mob metabolic rau 24 teev, thiab cov zis ntim thiab cov dej haus tau ntsuas [14]. Cov zis ntau ntawm creatinine (Cre), urea nitrogen (UN), calcium (Ca), Na-K-Cl, thiab qabzib (Glu) tau ntsuas nrog Dri Chem 3500V (FUJIFILM, Tokyo, Nyiv). Cov protein ntau hauv cov zis tau ntsuas siv Protein Assay Rapid Kit Wako II (FUJIFILM Wako Pure Chemical Corporation, Tokyo, Nyiv). Cov ntshav tau sau los ntawm caudal vena cava nyob rau hauv isoflurane anesthesia, thiab plasma concentrations ntawm UN, albumin (Alb), tag nrho cov protein (TP), tag nrho cov cholesterol (Tcho), Ca, thiab Na K-Cl raug ntsuas siv Dri-Chem 3500V. Plasma concentration ntawm Cre yog ntsuas siv FUJIFILM VET Systems Co., Ltd. (Tokyo, Nyiv). Creatinine (Ccre), qabzib (Cglu), calcium (Cca), sodium (Can), poov tshuaj (Ck), thiab chlorine (Ccl) clearances raug xam siv cov mis:
Clearance (Cx) (ml/min/kg)=[ zis concentration (mg/dl los mEq/l) × zis ntim (ml/min)] / plasma concentration (mg/dl lossis mEq/l) / lub cev hnyav (kg).
Fractional excretion ntawm qabzib (FEglu), calcium (FEca), sodium (FEna), poov tshuaj (FEk), thiab chlorine (FEcl) tau suav nrog cov mis:
Fraction excretion (FEx) (%)=Cx (ml/min/kg) / Ccre (ml/min/kg) × 100.
Kev ntsuas cov zis ntawm daim siab fatty-acid-binding (L-FABP) tau ntsuas siv nas / nas FABP1 / L-FABP cov khoom siv (R&D Systems, Inc., MN, USA)
Histology, immunofluorescence, thiab xam qhov chaw ntawm insulin-zoo
Tsiaj txhu tau euthanized nrog kev noj ntau dhau ntawm pentobarbital sodium tom qab 12 lub lis piam kho mob. Covlub raumthiab pancreas raug tshem tawm ntawm txhua tus nas euthanatized thiab hnyav. Cov qauv ntaub so ntswg tau kho nrog 4 feem pua paraformaldehyde nyob rau hauv phosphate-buffered saline (PBS), embedded nyob rau hauv paraffin raws li cov txheej txheem txheem, thiab sectioned ntawm 1 μm los yog 4 μm thickness rau periodic acid-Schiff (PAS) thiab immunofluorescence staining. Lub raum tubular dilation tau qhab nia raws li tau piav ua ntej [17]. Kaum random tsis-overlapping 40 x magnified teb ntawm lub raum cortex thiab ntawm juxtamedullary cheeb tsam tau qhab nia. Tubular dilation tau qhab nia ntawm qhov ntsuas ntawm {{10}}}-3, nrog 0, 1, 2, thiab 3 qhia 0,<5, 5–10,="" and="">10 dilated tubules per field, respectively. Pancreas sections were blocked with 3% bovine serum albumin (BSA) in PBS and incubated overnight at 4˚C with guinea pig anti-insulin polyclonal antibody (1: 100, Abcam K. K., Tokyo, Japan) or mouse anti-glucagon polyclonal antibody (1: 200, Abcam K.K.). The sections were rinsed with PBS and incubated for one hour at room temperature with Alexa Fluor 488 conjugated goat anti-guinea pig IgG antibody (1: 1000, Life Technologies, Carlsbad, CA) or Alexa Fluor 568 conjugated donkey anti-mouse IgG antibody (1: 1000, Life Technologies). After rinsing in PBS, the sections were mounted with ProLong Gold Antifade Reagent with DAPI (Life Technologies, Carlsbad, CA). Images were acquired with a Biozero BZ-X800 all-in-one fluorescence microscope, (KEYENCE, Tokyo, Japan). Islet areas and insulin-positive areas were measured in >10 ntu islet ib nas siv Image J software, thiab feem pua ntawm cov tshuaj insulin-zoo rau ib cheeb tsam islet raug xam.
Cov pob txha densitometry tsom xam ntawm femurs
Femurs raug excised los ntawm plaub euthanatized non-DM tswj, peb DM-cont, thiab peb DM-empa nas tom qab 12 lub lis piam kev kho mob thiab kuaj nrog ib tug micro-CT system (Latheta LCT-100, Aloka, Tokyo, Nyiv). . Pob txha pob txha pob txha ntom ntom (BMD, mg / cm3) thiab cov pob txha ntxhia cov ntsiab lus (BMC, mg) tau suav nrog Latheta software.
Gas chromatography-mass spectrometry (GC-MS) tsom xam ntawm plasma amino acids
Cov qauv ntshav tau txais los ntawm 30-cov nas tswj hnub nyoog ib asthiv thiab los ntawm 27–32 lub lis piam-laus DM cont thiab DM-empa nas tom qab 12-lub lim tiam kev kho mob. Cov plasma kuaj tau raug kho nrog Phenomenex EZ: ceev AA tsom xam cov khoom siv (Phenomenex, Torrance, CA), nrog norvalin siv raws li cov qauv sab hauv [18]. GCMS-QP2010Plus thiab GCMS daws software (Shimadzu, Kyoto, Nyiv) tau siv rau GC-MS tsom xam [19].
Kev txheeb cais
Cov txiaj ntsig, nthuav tawm raws li qhov txhais tau tias ± tus qauv yuam kev (SE), tau muab piv rau hauv ob pawg los ntawm cov tub ntxhais kawm ob-tailed t-tests thiab hauv ntau pawg los ntawm kev xeem Tukey-Kramer. P-tus nqi < 0.05="" tau="" suav="" tias="" yog="" qhov="" tseem="">
Cov txiaj ntsig
Cov xwm txheej dav dav cuam tshuam nrog ntshav qab zib
Kev ntsuas txhua lub lim tiam ntawm kev noj zaub mov pom tau tias qhov kwv yees qhov nruab nrab txhua hnub noj zaub mov tau siab dua me ntsis hauv DM-cont dua li hauv pawg DM-empa (36.9±1.5 g vs. 34.3±1.0 g). Qhov kwv yees noj txhua hnub ntawm empagliflozin hauv pawg DM-empa yog 23.7 ±{13}}.7 mg / kg lub cev qhov hnyav (BW). Empagliflozin txo cov ntshav qabzib ntau hauv ib hnub tom qab pib kho, los ntawm 404.6 ± 36.3 mg / dl rau 143.8 ± 14.6 mg / dl), nrog rau cov ntshav qabzib hauv DM-empa pawg tau qis dua hauv DM. -cont pab pawg thaum lub sijhawm 12-lub lim tiam kev kho mob (Fig 1A). Qib no hauv DM-empa tau zoo ib yam li kev tswj hwm ib txwm noj (147.{{40}}± 1.2 mg/dl) [14]. BW maj mam nce nrog hnub nyoog hauv pawg DM-empa, tab sis tsis nyob hauv pawg DM-cont (Daim duab 1B), nrog rau qhov kawg BW tom qab 12 lub lis piam yog 444.6 ± 18.0 g hauv DM empa thiab 372.3 ± 26.4 g hauv DM- pab pawg. Nyob rau tib lub hnub nyoog, lub cev hnyav ntawm kev tswj yog kwv yees li 533.4 ± 9.8 g [14]. Raws li lub cev qhov hnyav nce, adeps reins (1.4 ± 0.2 g) tseem nyob hauv DM-empa tab sis tsis nyob hauv pawg DM-cont.
Cov nas cuam tshuam DEK yav dhau los tau pom tias pom muaj cov piam thaj tsis txaus ntseeg, tsis muaj cov tshuaj insulin tso tawm hauv kev teb rau cov piam thaj, thiab poob ntawm cov hlwb [14]. Peb tau tshuaj xyuas seb empagliflozin puas txhim kho cov piam thaj thiab cov tshuaj insulin tso tawm thiab tiv thaiv kev poob ntawm cov hlwb. Cov ntshav qabzib tom qab 16 teev ntawm kev yoo mov tau qis dua hauv DM-empa ntau dua li hauv pawg DMcont, zoo li cov ntshav qabzib qib 60-240 min tom qab kev thauj khoom qabzib (Daim duab 1C). Lub sijhawm hloov pauv ntawm cov ntshav qabzib hauv OGTT ntawm DM-empa yuav luag zoo ib yam li cov nas uas tsis muaj ntshav qab zib ntawm DEK strain [14]. Txawm li cas los xij, glucose induced-insulin secretion tsis tshwm sim hauv ob pawg (Daim duab 1D). Immunofluorescent staining ntawm insulin nyob rau hauv pancreatic islets pom tsis muaj lossis faint insulin signals hauv pawg DM-cont thiab yuav luag 100 feem pua cov cim zoo hauv cov nas tswj, qhov tsis muaj zog insulin teeb liab, suav nrog 24.2 ± 3.5 feem pua ntawm ib cheeb tsam islet tau pom hauv peb nas nyob hauv DM-empa pawg (Fig 2).
Metabolic thiab lub raum tsis ua haujlwm thiab cov pob txha densitometry
Kev ntsuas ntawm metabolic tsis tau tom qab 12 lub lis piam kev kho mob tau pom tias kev noj zaub mov tsawg tsawg hauv DM-empa dua li hauv pawg DM-cont hauv ob qho tib si txheeb ze (Fig 3A) thiab qhov tseeb (S1 Fig). Tsis tas li ntawd, dej kom tsawg (Fig 3B, S1 Fig) thiab cov zis ntim (Daim duab 3C, S1 Fig) yog li 50 feem pua hauv DM-empa qis dua hauv pawg DM-cont.
Thaum kawg ntawm 12-lub lim tiam kev kho mob, plasma Tcho, Cre, Alb, TP, thiab Na concentrations tau qis dua hauv DM-cont dua li hauv pawg tswj hwm, nrog rau feem ntau ntawm cov kev txo qis hauv DM-cont pab pawg uas qhia ib nrab rov qab los ntawm pawg DM-empa. Tshwj xeeb, plasma Tcho, TP, thiab Na concentrations tau nce siab dua hauv DM-empa dua li hauv pawg DM-cont, txawm hais tias tag nrho cov concentrations qis dua hauv DM-empa dua li hauv pawg tswj hwm. Plasma Alb thiab Cre qib kuj tseem siab dua hauv DM-empa dua li hauv pawg DM-cont, tab sis cov kev sib txawv no tsis tseem ceeb (Table 1). Cov zis tso zis ntawm cov piam thaj, UN, Ca, Na, K, Cl, thiab cov protein ntau dua hauv DM-cont dua li hauv pawg tswj hwm, nrog rau feem ntau ntawm cov kev nce ntxiv hauv pawg DM-empa. Tshwj xeeb, cov zis tawm ntawm Glu, UN, Na, K, thiab Cl tau qis dua hauv DM-empa dua li hauv pawg DM-cont, txawm hais tias tag nrho cov excretions no tau nce siab dua hauv DM-empa dua li hauv pawg tswj hwm. . Kev tso zis ntau ntxiv ntawm Ca thiab cov protein kuj tau txo qis hauv pawg DM-empa, tsis muaj qhov sib txawv ntawm DM-empa thiab pawg tswj hwm (Table 2). Tsuas yog Ccre, uas tsis txawv qhov txawv ntawm peb pawg, tag nrho cov kev tshem tawm thiab cov seem excretions kuaj tau ntau dua hauv DM-cont dua li hauv pawg tswj hwm (Table 3). Ntxiv mus, tshwj tsis yog rau Cglu thiab FEglu, uas tseem siab dua hauv pawg DM-empa, feem ntau ntawm cov nce hauv pawg DM-cont tau txo qis hauv pawg DM-empa. Tshwj xeeb, Cca, Cna, thiab Ccl tau qis dua hauv DM-empa dua li hauv pawg DM-cont tab sis tseem nyob hauv DM-empa ntau dua li hauv pawg tswj hwm (Table 3).
Femoral BMD tau qis dua hauv DM-cont dua li hauv pawg tswj hwm, nrog empagliflozin pom los kho femoral BMD hauv pawg DM-empa (Table 4). BMC nyiam siab dua hauv DM-empa dua li hauv pawg DM-cont, tab sis qhov sib txawv tsis tseem ceeb (Table 4)
Pathology ntawm lub raum
Cov nas cuam tshuam DEK tau nthuav tawm lub raum loj zuj zus nrog cov tubules dilated [14]. Txawm hais tias kev kho empagliflozin txo qis cov ntshav qabzib ntau, nws tsis txo qis
lub raum hnyav, uas zoo sib xws hauv pawg DM-empa thiab DM-cont (2.35 ± 0 08 g vs. 2.24 ± 0.08 g). Kev kuaj mob histologic tau pom tias qhov loj me me yog qhov tseem ceeb hauv lub raum tubules ntawm thaj chaw juxtamedullary hauv ob qho tib si DM-cont thiab DM-empa nas (Fig 4A). Empagliflozin tsis txo qis qhov dilation hauv lub raum tubules (Daim duab 4B thiab 4C). Raws li qhov no, kev tso zis ntawm L-FABP, biomarker ntawm lub raum tubular raug mob [20], tau siab dua hauv DM-cont thiab DM-empa piv rau Kev Tswj, thiab tsis muaj qhov sib txawv ntawm DM-cont thiab DM-empa ( Daim duab 4D thiab 4E). Raws li yav dhau los qhia [14], tsis muaj glomerular lesion xws li hypertrophy hauv Control, DM-cont, thiab DM-empa (Fig 4F).
GC-MS tsom xam ntawm plasma amino acids
Metabolic profiling using PLS-DA multivariate analysis showed differences in amino acid status among the control, DM-cont, and DM-empa groups (S2 Fig). The loading plot corresponded to a score plot consisting of 24 amino acids (S2 Fig). The variable influences on projection (VIP) parameters are shown in S2 Fig. Amino acids colored red had VIP >1 and were considered to have a relatively high impact for separating these groups. Moreover, statistical analysis of amino acids showed a VIP >1.
Plasma sarcosine zoo li siab dua hauv DM-cont dua li hauv pawg tswj hwm, nrog empagliflozin ntxiv cov ntshav sarcosine ntxiv. Hauv qhov sib piv, plasma tryptophan zoo li qis dua hauv DM-cont dua li hauv pawg tswj hwm, nrog empagliflozin ntxiv txo cov ntshav tryptophan (Table 5). Plasma glycine qib tau zoo sib xws hauv DM-cont thiab tswj pawg tab sis tau nce siab dua hauv DM-empa dua li ntawm ob pawg. Glutamine thiab ornithine concentrations tau qis dua hauv DM-cont dua li hauv pawg tswj hwm thiab tsis tau rov qab los ntawm kev kho empagliflozin. Piv nrog rau pawg tswj hwm, plasma -aminobutyric acid tau qis dua hauv pawg DM-cont, nrog empagliflozin ntxiv txo nws cov concentration. Plasma valine thiab isoleucine concentrations tau nce siab dua hauv DM-cont dua li hauv pawg tswj hwm, nrog rau cov concentrations rov qab mus rau qib tswj los ntawm kev kho empagliflozin.
Kev sib tham
SGLT2 inhibitors txo cov ntshav qabzib hauv ib qho kev ywj pheej ntawm insulin thiab qhia txog kev tiv thaiv kev tiv thaiv hauv cov neeg mob uas muaj hom 2 DM, nrog rau ntau tus tsiaj qauv [8, 12, 21–23]. Feem ntau cov kev tshawb fawb tsiaj nrog SGLT2 inhibitors tau ua rau cov tsiaj rog rog nrog hom 2 DM [22, 24-28]. Txawm li cas los xij, ntau hom 2 DM cov neeg mob, tshwj xeeb tshaj yog nyob rau hauv Asia, tsis rog [29]. Cov teebmeem ntawm SGLT2 inhibitors rau cov neeg mob lub raum mob lossis cov ntshav qab zib tsis rog tseem tsis meej [29, 30]. Txoj kev tshawb fawb tam sim no tau tshawb xyuas qhov cuam tshuam ntawm empagliflozin ntawm cov metabolism, lub raum kev ua haujlwm, thiab histopathology hauv DEK nas, tus qauv tshiab uas tsis yog rog rog nrog rau kev ua rau lub raum loj [14]. Piv nrog rau cov tsiaj mob ntshav qab zib mellitus nrog cov rog rog, cov phenotype ntawm cov kab mob ntshav qab zib DEK nas tau tshwm sim los ntawm kev txo qis ntawm cov tshuaj insulin hauv cov ntshav vim qhov poob ntawm cov hlwb pancreatic es tsis yog insulin tsis kam, yog li cov nas ntshav qab zib DEK pom qhov txo qis hauv BW thiab rog nrog rau cov kab mob pancreatic. kev loj hlob ntawm DM. Tsis tas li ntawd, cov kab mob ntshav qab zib DEK tau pom tias muaj zog rau lub raum parenchyma thiab ua rau lub raum loj dua nrog rau lub raum tubules [14].
SGLT2 inhibitors txhawb kev siv cov rog, cov leeg, thiab cov amino acids ua lub zog [31–33], txo cov rog thiab BW hauv tib neeg rog thiab tsiaj txhu [28, 34, 35]. Kev poob ntawm cov rog thiab txo qhov hnyav nce kuj tau pom nyob rau hauv cov nas uas tsis rog ntshav qab zib tau kho nrog SGLT2 inhibitors [31]. Tsis tas li ntawd, tib neeg thiab cov nas uas tau kho nrog SGLT2 inhibitors tau pom tias muaj kev them nyiaj hyperphagia los tswj lawv lub zog thiab cov khoom noj sib npaug [27, 32, 36]. Hauv txoj kev tshawb no, kev kho empagliflozin txo qis cov piam thaj hauv cov ntshav ntawm DEK nas nyob rau hauv 24 teev, nrog rau cov kev txo qis no tau khaws cia thoob plaws hauv 12-lub lim tiam kev kho mob, zoo ib yam li lwm cov kev tshawb fawb ntawm SGLT2 inhibitors [15, 24]. Hauv qhov sib piv rau qhov kev tshawb pom yav dhau los [27, 28,36], peb pom tias kev kho mob empagliflozin txo qis kev noj zaub mov thiab txo qis cov adeps reins thiab inhibition ntawm BW nce. Qhov tshwm sim tsis tau npaj tseg no yog tej zaum cuam tshuam nrog kev txhim kho hauv lub zog sib npaug. Thaum pib, cov piam thaj tso zis tso zis tau qis dua hauv DM-empa dua li DM-cont nas, txawm hais tias Cglu siab dua hauv pawg DM empa tau tshwm sim los ntawm kev ua tiav inhibition ntawm qabzib reabsorption, nrog FEglu yuav luag 100 feem pua. Qhov kev tshawb pom no tau txhawb ntxiv los ntawm kev txo qis hauv cov zis UN tso tawm, qhov ntsuas ntawm cov protein catabolism, txawm tias ib txwm BUN qib, hauv pawg DM-empa [37]. Tsis tas li ntawd, qhov nce hauv plasma TP thiab txo qis hauv cov zis protein hauv DM-empa pawg qhia tau hais tias empagliflozin txhim kho cov protein tuav. Kev khaws cia thiab kev siv cov piam thaj thiab kev thim rov qab los ntawm catabolic mus rau lub xeev assimilated hauv cov nas uas kho nrog empagliflozin tuaj yeem yog vim nws tswj tau cov ntshav qabzib zoo.
SGLT2 inhibitors tau pom tias ua kom cov zis ntim, nrog rau kev nce hauv tubular reabsorption compensating rau poob hauv lub cev kua thiab electrolytes [21, 25, 38]. Yog li ntawd, cov zis ntim tau nce tam sim ntawd tom qab pib ntawm SGLT2 inhibitor kev kho mob, tab sis kev kho mob ntev yuav txo tau cov zis ntim thiab dej kom tsawg [38, 39]. Txawm hais tias plasma Cre qib tau qis dua hauv DM-cont thiab DM-empa dua li hauv Pawg Tswjhwm, Ccre tsis txawv ntawm cov tswj, DM-cont, thiab DM-empa pawg. Yog li ntawd, txij li tsis muaj pov thawj pom tseeb ntawm glomerular hypertrophy hauv DEK-DM nas [14], glomerular hyperfiltration yuav nyuam qhuav tshwm sim los yog mob me me hauv DM-cont. Tsis tas li ntawd, lub raum loj nrog cov parenchyma nce ntxiv tuaj yeem ua rau muaj tus lej nephron ntau ntxiv, qhia tias lub raum loj hauv ob qho tib si DM-cont thiab DM-empa tuaj yeem cuam tshuam txog kev txo cov ntshav ntshav Cre. Yog li ntawd, qhov txo cov zis ntim hauv pawg DM-empa tuaj yeem tshwm sim los ntawm kev txhim kho cov dej reabsorption hauv lub raum tubules, tsis yog los ntawm qhov txo glomerular pom tus nqi. Zuag qhia tag nrho, kev tso zis ntau ntxiv ntawm electrolytes hauv pawg DM-cont tau txo qis hauv DM empa pawg, qhia tias txo cov ntshav plasma ntau ntawm electrolytes hauv DM-cont pawg tau txo qis los ntawm kev kho empagliflozin los ntawm kev txhim kho lub raum tubular muaj nuj nqi. Qhov kev xav no tau txhawb nqa los ntawm kev txo qis ntawm lawv cov kev tshem tawm nrog kev txo qis qis hauv cov pab pawg DM-empa.
SGLT2 inhibitors tau tshaj tawm tias muaj kev cuam tshuam rau cov pob txha [31, 40]. Vim tias osteoporosis yog ib qho teeb meem ntawm DM, peb piv BMD thiab BMC hauv peb pawg nas. BMD thiab BMC tau qis dua hauv DM-cont dua li hauv pawg tswj hwm, tab sis cov kev txo qis no tau txo qis hauv pawg DM-empa. Ntxiv mus, cov kev nce hauv BMD thiab BMC hauv pawg DM-empa tau nrog los ntawm kev txo qis hauv cov zis Ca excretion. Vim tias cov qib plasma Ca tau tswj hwm nruj, tsis muaj qhov sib txawv tseem ceeb hauv plasma Ca qib ntawm cov tswj, DM-cont, thiab DM-empa pawg. Cov kev tshawb pom no tau pom tias qhov tsis zoo Ca tshuav nyiaj li cas hauv pawg DM-cont tuaj yeem raug cuam tshuam los ntawm kev sib koom ua ke ntawm cov pob txha mus rau hauv cov hlab ntsha, nrog rau qhov xwm txheej no raug txo qis los ntawm kev txhim kho lub raum tubular ua haujlwm ntawm Ca hauv pawg DM-empa.
Kev txo qis ntawm hypoproteinemia hauv pawg DM-empa tuaj yeem tshwm sim los ntawm kev txhim kho cov proteinuria. Txawm hais tias glomerular hyperfiltration thiab hypertrophy tsis tau pom meej meej hauv pawg DM-cont, txo cov ntshav qabzib tuaj yeem txo qis glomerular pom Alb. Tsis tas li ntawd, kev txhim kho hauv tubular reabsorption ntawm cov protein thiab cov amino acids tuaj yeem pab txhawb rau kev sib txuas ntawm plasma proteins hauv daim siab. Qhov cuam tshuam DEK nas yog tus cwj pwm los ntawm cov ntshav plasma Tcho tsawg [14]. Vim tias kev tsim tawm ntau ntxiv ntawm Alb yog nrog los ntawm kev tsim cov Tcho hauv daim siab ntau dua [41], ntau dua plasma Tcho qib hauv pawg DM-empa tuaj yeem cuam tshuam nrog kev nce siab hauv siab ntawm Alb. Cov tom kawg tuaj yeem tshwm sim los ntawm kev nce cov amino acid thiab cov khoom siv hluav taws xob ntsig txog kev txhim kho cov protein tuav thiab lub zog sib npaug.
Branched-chain amino acid (BCAA) homeostasis feem ntau txiav txim siab los ntawm BCAA catabolic kev ua haujlwm hauv cov ntaub so ntswg [42], nrog qhov tsis xws ntawm BCAA homeostasis suav tias yog qhov ntsuas ntawm insulin tsis kam lossis mob ntshav qab zib [43-45]. Peb qhov kev soj ntsuam GC-MS ntawm cov amino acids tau pom tias plasma concentrations ntawm valine thiab isoleucine hauv DM-empa nas sib npaug rau cov nas uas tsis muaj ntshav qab zib, qhia tias kev kho empagliflozin ameliorated protein catabolism thiab mob ntshav qab zib. Tsis tas li ntawd, peb tau pom tias qhov nce ntawm empagliflozin tshwj xeeb hauv plasma glycine qib tau cuam tshuam nrog kev txhim kho ntshav qab zib. Qhov zoo siab, qib qis glycine hauv cov ntshav ntshav qis tau pom hauv tib neeg mob ntshav qab zib [46], thiab kev tswj hwm glycine tau pom los ua kom muaj cov tshuaj tiv thaiv oxidative thiab renoprotective hauv cov nas uas muaj streptozotocin-induced diabetes [47]. Cov qib siab glycine ntau dua hauv plasma yuav cuam tshuam nrog kev txo qis ntawm oxidative kev nyuaj siab thiab mob ntshav qab zib nephropathy hauv cov neeg mob ntshav qab zib thiab cov tsiaj sim kho nrog empagliflozin. Plasma qib ntawm cov amino acids raug cuam tshuam los ntawm lawv kev siv los ntawm cov ntaub so ntswg thiab cov kabmob thoob plaws hauv lub cev thiab nyob ntawm cov mob anabolic thiab catabolic. Cov kev tshawb fawb ntxiv yuav tsum tau txiav txim siab seb qhov kev hloov pauv ntawm cov qib no thiab lwm cov amino acids cuam tshuam li cas rau cov mob metabolic.
SGLT2 inhibitors siv cov teebmeem renoprotective los ntawm inhibiting ntau cov txheej txheem cuam tshuam nrog rau cov kab mob hauv lub raum, xws li albuminuria thiab nce raum hnyav [15, 24, 48, 49]. Lub raum tubule dilation thiab proteinuria yog cov cim ntawm ntshav qab zib cuam tshuam rau lub raum puas [50]. Txawm li cas los xij, empagliflozin tsis tuaj yeem txwv tsis pub lub raum tubule dilation thiab nce tso zis ntawm L-FABP hauv peb cov qauv nas, tab sis txhim kho tubular muaj nuj nqi. Qhov sib txawv no tau pom tias qhov kev txhim kho ntawm empagliflozin-txuas rau hauv tubular muaj nuj nqi tshwm sim los ntawm cov teebmeem me lossis theem nrab tsis nrog cov kev hloov pauv ntawm cov organic hauv lub raum tubules. Xwb, vim hais tias tubular dilation nrog kev raug mob thiab nce lub raum hnyav tau khaws cia nyob rau hauv cov xwm txheej normoglycemic nyob rau hauv cov nas cuam tshuam DEK, cov kev hloov no tej zaum yuav yog cov caj ces muaj feem xyuam rau cov ntshav qab zib hauv DEK nas es tsis yog theem nrab rau hyperglycemia.
Txawm hais tias SGLT2 inhibitors tsom rau lub raum tubules, me ntsis paub txog qhov cuam tshuam ntawm cov neeg ua haujlwm ntawm tag nrho cov kev hloov pauv hauv kev ua haujlwm ntawm lub raum tubules, tshwj tsis yog tias glomerular hypertrophy cuam tshuam nrog tubular cotransport ntawm sodium nrog piam thaj [13, 22, 24]. Kev tshawb fawb ntawm cov teebmeem ntawm SGLT2 inhibitors ntawm tubular muaj nuj nqi tau pom tias cov dej reabsorption tsis yog tsuas yog suppressed los ntawm osmotic diuresis tab sis tau txhim kho los ntawm upregulation ntawm aquaporin thiab urea transporter los ntawm ib qho tseem tsis tau txiav txim siab [21, 25]. Lub raum tubular epithelial hlwb ntawm cov neeg mob ntshav qab zib tau cuam tshuam tsis yog los ntawm osmotic siab ntawm cov piam thaj tam sim no hauv cov kua dej tab sis los ntawm cov piam thaj ntau dhau thiab sodium coj mus rau hauv cov hlwb ntawm SGLT2. Tsis ntev los no, oxidative kev nyuaj siab nyob rau hauv lub raum tubules tau suav hais tias yog ib qho tseem ceeb rau kev loj hlob ntawm mob ntshav qab zib nephropathy (DN), thiab ipragliflozin pom renoprotective los ntawm kev tiv thaiv overproduction ntawm ROS nyob rau hauv lub raum tubules [51]. Txawm hais tias tsis muaj qhov tshwm sim loj ntawm DN tau pom nyob rau hauv cov nas cuam tshuam DEK, oxidative kev nyuaj siab yuav ua rau lub cev tsis zoo. Tam sim no, tsis muaj kev pom zoo sib koom ua ke txog qhov cuam tshuam ntawm SGLT2 inhibitors ntawm tubular muaj nuj nqi, qhia tias yuav tsum muaj kev tshawb fawb ntxiv los piav qhia cov txheej txheem uas empagliflozin txhim kho tubular muaj nuj nqi hauv DEK nas.
SGLT2 inhibitors tau pom los khaws cov cell loj hauv hom 1 DM thiab rog hom 2 DM qauv [52, 53]. Peb kuaj pom cov hlwb uas tsis muaj zog insulin hauv pawg DM-empa, tab sis tsis muaj cov hlwb hauv pawg DM-cont. Txawm li cas los xij, OGTT tau pom tias muaj kev txhim kho me ntsis hauv cov piam thaj uas tsis muaj insulin tso tawm hauv pawg DM-empa. Vim tias empagliflozin muaj ib nrab-lub neej luv luv ntawm 8 teev, qhov txo cov ntshav qabzib tau pom tom qab 16 teev nrawm thiab kev txhim kho hauv cov piam thaj tsis txaus ntseeg vim qhov cuam tshuam ntawm cov seem empagliflozin. Cov kev tshawb pom no, yog li ntawd, qhia tias kev tswj hwm mus sij hawm ntev ntawm empagliflozin tuaj yeem txhim kho cov piam thaj metabolism thiab txo qis cov piam thaj hauv cov ntshav tom qab txhaj tshuaj ib zaug hauv ib qho kev ywj pheej ntawm insulin.
Hauv kev xaus, cov txiaj ntsig ntawm txoj kev tshawb fawb tam sim no qhia tias empagliflozin ameliorates cov metabolism hauv lub cev thiab lub raum ua haujlwm hauv DEK nas, txawm hais tias cov txheej txheem ntxaws hauv qab cov lus teb tshwj xeeb ntawm cov tsiaj no rau empagliflozin tseem tsis meej. Cov kev tshawb pom no tseem qhia tau tias DEK nas yog ib qho qauv zoo rau kev kawm thiab tshuaj xyuas ntau yam teebmeem ntawm SGLT2 inhibitors hauv ntshav qab zib.
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