Cov teebmeem ntawm Saponin Los Ntawm Cov Noob Ntawm Litchi Chinensis Sonn Ntawm TGF- 1, FN Thiab SOCS-1 nyob rau hauv lub raum Tubular Epithelial Cells nyob rau hauv siab Glucose

Hu rau:joanna.jia@wecistanche.com/ WhatsApp: 008618081934791

Hai-Yang Nie, Rui Chen, Hong-Na Zhang, Zhi Pan

1 Changchun University of Chinese Medicine, Changchun, Suav.

2 Shunyi Tsev Kho Mob ntawm Beijing Suav Tshuaj Kho Mob, Beijing, Suav.

Qhov tseem ceeb

Saponin los ntawm cov noob ntawm Litchi chinensis Sonn txo cov apoptosis ntawm lub raum tubular epithelial hlwb thiab tso tawm ntawm TGF- 1 thiab FN.

Abstract

Lub Hom Phiaj: Tshawb xyuas cov txiaj ntsig ntawm saponin los ntawm cov noob ntawm Litchi chinensis Sonn (SLS) ntawm kev loj hlob thiab apoptosis ntawmtib neegraumepithelial hlwb (HKC) kab lis kev cai hauv cov piam thaj siab. Cov txheej txheem: HKC tau coj mus kuaj hauv DMEM / F12 nruab nrab ntxiv nrog 30 mmol / L qabzib thiab kho nrog lossis tsis muaj SLS. Hauv pawg ib txwm muaj, ib qho isometric DMEM / F12 nruab nrab nrog 5.5mmol / L qabzib tau ntxiv. Kev zais ntawm TGF- 1 thiab fibronectin (FN) tau kuaj pom los ntawm ELISA. Cell apoptosis tau kuaj pom los ntawm txoj kev Annexin V-FITC / PI ob chav staining. Western blot tau siv los txheeb xyuas cov theem ntawm kev tawm tsam ntawm cytokine signaling-1 (SOCS-1).

Cov txiaj ntsig: Cov txiaj ntsig ntawm ELISA tau pom tias qhov tso tawm ntawm TGF{{0}}} thiab FN tau txo qis hauv SLS pawg piv nrog cov pab pawg hauv 30 mmol / L qabzib (P < {{="" 9}}="" 05).="" muaj="" ntau="" lub="" hlwb="" apoptosis="" nyob="" rau="" hauv="" 30="" mmol="" l="" qabzib="" pab="" pawg="" dua="" li="" hauv="" pawg="" ib="" txwm="" (p=""><0.01). piv="" nrog="" rau="" 30="" mmol="" l="" glucose="" kho="" pab="" pawg,="" apoptosis="" ntawm="" hkc="" tau="" txo="" qis="" hauv="" sls="" pawg="" (p=""><0.01). western="" blot="" tau="" pom="" tias="" qib="" socs{11}}="" hauv="" cov="" piam="" thaj="" siab="" ntxiv="" rau="" sls="" pawg="" tau="" txo="" qis="" (p="">< 0.01),="" piv="" nrog="" cov="" piam="" thaj="">

Cov ntsiab lus: SLS tuaj yeem txo qhov tso tawm ntawm TGF- 1 thiab FN hauv HKC los ntawm kev txo cov deposition ntawm extracellular matrix. SLS kuj txo qis apoptosis ntawm HKC los ntawm inhibiting qib ntawm SOCS-1. Cov txiaj ntsig no qhia txog lub luag haujlwm ntawm SLS hauv kev tiv thaiv kev nce qib ntawm glomerular sclerosis.

Keywords: Saponin, Noob of Litchi chinensis Sonn, Glucose,Lub raumepithelial hlwb, TGF- 1, Fibronectin, Apoptosis

cistanchekho taumob raumtxhim kholub raum ua haujlwm

Keeb kwm

Mob ntshav qab zib nephropathy (DN) yog ib qho teeb meem loj tshaj plaws ntawm cov ntshav qab zib mellitus nrog qhov tshwm sim siab thiab ua rau tuag taus, tus cwj pwm los ntawm glomerular sclerosis thiab lub raum interstitial fibrosis. Tib neegraumepithelial hlwb (HKC) yog lub hom phiaj tseem ceeb hauv DN thiab lawv zais ntau cov cytokines, xws li TGF- 1 thiab fibronectin (FN), uas ua rau muaj qhov tshwm sim ntawm glomerulosclerosis thiab lub raum interstitial fibrosis. Lub caij no, nws tau raug tshaj tawm tias cov piam thaj siab tuaj yeem ua rau HKC apoptosis nce qib ntawm kev txo qis ntawm cytokine signaling-1(SOCS-1)[1,2]. Yog li ntawd, inhibiting qib ntawm TGF- 1, FN thiab SOCS-1 yog qhov tseem ceeb rau kev tiv thaiv kev nce qib ntawm glomerular sclerosis. Cov noob ntawm Litchi chinensis Somm (SLS), yog cov tshuaj suav suav feem ntau faib hauv Guangdong. Fujian thiab Guangxi xeev ntawm Tuam Tshoj tau sau thawj zaug hauv Bencaogangmu hauv Ming Dynasty ntawm Tuam Tshoj (1518 AD-1593 AD). Saponin yog cov khoom xyaw nquag ntawm cov noob ntawm Litchi chinensis Sony. Raws li peb txoj kev tshawb fawb yav dhau los, txoj kev tshawb fawb no tshawb fawb ntxiv txog cov txiaj ntsig ntawm SLS ntawm qib TGF- 1 thiab FN hauv HKC kab lis kev cai nrog cov qabzib siab, apoptosis ntawm HKC, thiab lub hauv paus txheej txheem ntawm kev tiv thaiv thiab kho glomerulosclerosis [ 3].

cistanchekho taumob raum tsis ua haujlwm

Cov ntaub ntawv thiab cov txheej txheem Cov ntaub ntawv

SLS nrog purity ntau dua 98 feem pua ​​​​tau yuav los ntawm Xianqingle Biological Co., LTD (Xi'an, Tuam Tshoj).HKC tau yuav los ntawm Shanghai Fuxiang Biotechnology Co., Ltd (Shanghai, Tuam Tshoj). DMEM / F12 nruab nrab, tus qauv fetal bovine serum, thiab trypsin yog los ntawm Hyclone (USA). Lub dimethyl subfamily yog los ntawm Sigma (USA). Cov khoom siv ELISA tau yuav los ntawm Shanghai Longton Biotechnology Company (Shanghai, Suav). Annexin V-FITC/PI Apoptosis Detection Kit yog los ntawm Nanjing Kaiji Biotechnology Co., Ltd (Nanjing, Suav).

Cells kab lis kev cai

HKC tau khaws cia hauv DMEM/F12 nrog 10 feem pua ​​​​ntawm cov fetal calf serum thiab tau coj mus kuaj ntawm 37 degree hauv cov huab cua 95 feem pua ​​​​ntawm huab cua thiab 5 feem pua ​​​​CO.Cells tau zom nrog 0.25 feem pua ​​trypsin tom qab 2-3 hnub siv.

Enzyme-linked immunosorbent assay

HKC tau muab faib ua rau pawg, suav nrog pawg tswj hwm (DMEM nrog 5.5mmo / L qabzib), pab pawg qabzib siab (DMEM nrog 30mmol / L qabzib), SLS pawg koob tshuaj tsawg (5mg / ml, S1 ntxiv rau 30mmol / L qabzib), SLS Cov koob tshuaj nruab nrab (10mg / ml, S2 ntxiv rau 30mmol / L qabzib), SLS pawg koob tshuaj siab (20mg / ml, S3 ntxiv rau 30mmol / Lglucose), thiab pab pawg neeg dawb paug (DMEM tsis muaj hlwb).

48 teev tom qab ntawd, cov supernatant ntawm txhua pab pawg tau sau, thiab cov ntsiab lus ntawm TGF- 1 thiab FN tau kuaj pom los ntawm ELISA. Kev ua haujlwm tau ua raws li phau ntawv qhia. Tus nqi OD tau nyeem ntawm 450nm los ntawm tus nyeem ntawv microplate. Qhov siab ntawm TGF- 1 thiab FN tau txiav txim siab raws li tus qauv nkhaus.

acteosidehauvcistanchemuaj cov teebmeem zoo rauraum

Flow cytometry

Cov pab pawg sim tau zoo ib yam li saum toj no. Cov hlwb apoptosis tau kuaj pom los ntawm kev ntws cytometry nrog Annexin V-FITC / PI ob chav staining. Sab laug sab laug sawv cev rau feem pua ​​​​ntawm cov hlwb necrotic, sab qaum txoj cai quadrant sawv cev rau feem pua ​​​​ntawm cov hlwb apoptosis lig, sab laug quadrant sawv cev rau feem pua ​​​​ntawm cov hlwb siv tau, thiab sab xis sab xis sawv cev rau feem pua ​​​​ntawm cov hlwb apoptosis thaum ntxov.

Western blot tsom xam

HKC tau kho nrog 10 * mol / L ntawm Angiotension (Ang) Ⅱ, 20mg / Lof SLS thiab 10 * mol / L ntawm AngⅡ, DMEM / F12 nruab nrab, 20mg / L ntawm SLS thiab 25mmol / L ntawm qabzib thiab 25mmol / L ntawm cov piam thaj. , raws.

Cell lysate proteins tau daws ntawm sodium dodecyl sulfate-polyacrylamide gel electrophoresis thiab xa mus rau PVDF daim nyias nyias. Txhua daim nyias nyias tau incubated nrog ib tug monoclonal antibody, ua raws li incubation nrog peroxidase-conjugated secondary antibodies thiab chemiluminescence nrhiav tau.

Kev txheeb cais

Txhua qhov kev txheeb xyuas txheeb cais tau ua tiav siv SPSS version 19.0. Qhov sib txawv ntawm pawg cov ntaub ntawv sim tau soj ntsuam los ntawm ib-txoj kev ANOVA, thiab qhov sib txawv tsawg kawg yog siv los ntawm cov pab pawg. P<0.05 was="" considered="" to="" indicate="" statistical="">

Cov txiaj ntsig

Kev cuam tshuam ntawm SLS ntawm kev tso tawm ntawm TGF-B1 thiab FN Piv nrog rau cov qib qabzib siab, SLS tuaj yeem cuam tshuam cov qib ntawm TGF-B1 thiab FN hauv HKC (P<0.01 or=""><0.05)(figure 1,="" figure="">

Piv nrog rau cov qib qabzib siab: * P<0.05, **=""><0.01. control="" group:="" dmem="" with="" 5.5mmol/l="" glucose,="" high="" glucose="" group:="" dmem="" with="" 30mmol/l="" glucose,="" s1="" group:="" 5mg/ml="" sls+30mmol/l="" glucose,="" s2="" group:="" 10mg/ml="" sls+30mmol/l="" glucose,="" s3="" group:="" 20mg="" l="" sls+30mmol/l="" glucose.="" sls,="" seed="" of="" litchi="" chinensis="">

Kev cuam tshuam ntawm SLS ntawm HKC apoptosis Qhov tshwm sim ntawm kev ntws cytometry tau pom tias apoptosis ntawm cov piam thaj ntau ntau dua li cov pab pawg ib txwm muaj (P < {{0}}}.01).="" piv="" nrog="" rau="" cov="" qib="" qabzib="" siab,="" qhov="" apoptosis="" tus="" nqi="" ntawm="" hkc="" tau="" cuam="" tshuam="" tsis="" zoo="" nrog="" cov="" ntsiab="" lus="" ntawm="" sls="" (p=""><0.01) (daim="" duab="">

Kev cuam tshuam ntawm SLS ntawm qib SOCS{{0}} Qib SOCS-1 cov protein nyob hauv pawg AngII tau nce siab dua li cov pab pawg ib txwm muaj (P < 0.="" {8}}1).="" qib="" ntawm="" socs-1="" hauv="" pawg="" angii="" ntxiv="" rau="" sls="" qis="" dua="" li="" hauv="" pawg="" angii="" tab="" sis="" tseem="" siab="" dua="" li="" hauv="" pawg="" tswj="" hwm="" (p="">< 0.01).="" piv="" nrog="" rau="" cov="" pab="" pawg="" ib="" txwm="" muaj,="" qib="" ntawm="" socs-1="" cov="" protein="" nyob="" rau="" hauv="" cov="" ntshav="" qabzib="" siab="" tau="" nce="" (p=""><0.01). piv="" nrog="" rau="" cov="" qib="" qabzib="" siab,="" qib="" protein="" ntawm="" cov="" piam="" thaj="" siab="" ntxiv="" rau="" sls="" pawg="" tau="" txo="" qis="" (p=""><0.01) (daim="" duab="">

Daim duab 4 Qhov cuam tshuam ntawm SLS ntawm kev qhia ntawm SOCS-1 a. AngII pawg: 10-6 mol/L Angiotension Ⅱ, b. AngII plus SLS pawg: 20mg/L SLS thiab 10-6 mol/L AngⅡ, c. Tswj pawg: DMEM / F12 nruab nrab, d. Cov piam thaj ntau ntxiv rau SLS pawg: 20mg / L SLS ntxiv rau 25mmol / L ntawm qabzib, e. Tswj pawg: DMEM / F12 nruab nrab, f. Cov piam thaj ntau: 25 mmol / L. SOCS-1, suppressor ntawm cytokine signaling-1. SLS, Noob ntawm Litchi chinensis Sonn.

Kev sib tham

Deposition ntawm extracellular matrix thiab apoptosis ntawm lub raum tubular epithelium yog lub ntsiab pathological hloov nyob rau hauv txoj kev loj hlob ntawm DN. Raws li ib txwm muaj, morphology thiab naj npawb ntawm HKC tseem nyob ruaj khov. Tab sis nyob rau hauv ntau yam stimulating yam xws li o, HKC tuaj yeem loj hlob tuaj [4]. Cov piam thaj ntau ua rau lub siab tso tawm ntawm TGF- 1 thiab FN ntawm HKC thiab ua kom cov apoptosis ntawm HKC, uas yog qhov tseem ceeb hauv kev txhawb nqa qhov tshwm sim ntawm DN. Hauv qhov kev sim no, cov piam thaj siab tau siv los ua kev txhawb nqa. Nws tau pom tias SLS tuaj yeem cuam tshuam qhov tso tawm ntawm TGF- 1, FN thiab txo HKC apoptosis los ntawm kev txo qis ntawm SOCS-1.

Litchi chinensis Sonn feem ntau yog muab faib rau hauv Guangdong, Fujian, thiab Guangxi xeev ntawm Tuam Tshoj. Nws tau tshaj tawm tias SLS muaj cov tshuaj tiv thaiv hyperglycemic, antihyperlipidemic, antiplatelet, thiab tshuaj tiv thaiv kab mob [5]. Raws li cov ntaub ntawv ntawm Bencaogangmu nyob rau hauv Ming Dynasty ntawm Tuam Tshoj (1518 AD - 1593 AD), SLS muaj peev xwm "kub nqhis dej". Nyob rau hauv xyoo tas los no, ntau tus kws tshawb fawb tau ua ntau yam kev tshawb fawb txog nws cov tshuaj pharmacological thiab tshuaj muaj pes tsawg leeg. Cov tshuaj tseem ceeb ntawm SLS yog cov khoom sib xyaw xws li lychee saponins, flavonoids, polyphenols, chisels, amino acids, fatty acids, thiab cov khoom tsis haum. SLS yog cov khoom xyaw tseem ceeb. SLS yog dav siv hauv ntshav qab zib thiab mob ntshav qab zib nephropathy, nrog rau cov tshuaj hypoglycemic, hypolipidemic, inhibition ntawm kev tiv thaiv kab mob o [6]. Cov kev tshawb fawb niaj hnub no kuj tau lees paub tias SLS muaj cov nyhuv hypoglycemic [7]. Txawm li cas los xij, cov txheej txheem ntawm cov nyhuv no tsis meej.

Hauv txoj kev tshawb no, theem ntawm TGF- 1 thiab FN hauv HKC tau nce siab tom qab stimulation nrog cov qabzib siab, thiab qib ntawm TGF- 1 thiab FN tau poob qis heev tom qab kev kho SLS nyob rau hauv ib koob tshuaj. . SOCS-1 qib protein tau txo qis tom qab kev kho SLS. Txoj kev tshawb no tau muab lub hauv paus theoretical rau kev kho mob ntawm SLS.

Cistanchetuaj yeem kho taumob raum

Cov ntaub ntawv

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2. Nishimoto N, Kishimoto T, Yoshizakik. Anti-interleukin6 receptor antibody kho nyob rau hauv tus kab mob rheumatic. Ann Rheum Dis 2000, 59(supple1): 121.

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