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集思慧遠客戶發表《醋灸處理甘遂對大鼠腸道微生物群的調節及解毒作用》

幸运飞艇官方开户FABURIQI:2019-06-11 LIULANCISHU:448

Euphorbia kansui fry-baked with vinegar modulates gut microbiota and reduces intestinal toxicity in rats

IF=3.49

合作單位:江蘇省中藥方劑高技術研究重點實驗室,

南京中醫藥大學

摘要

      民族藥理學相關:甘遂(EK),是一種有毒的中藥,經常被用來治療水腫、腹水和哮喘。用醋灸處理(VEK)可用于降低中藥的毒性。以往的研究證實,醋灸可顯著降低甘遂的胃腸道毒性。EK的毒副作用與腸道密切相關,但是現有的研究結果無法提供解毒的實用措施以及醋灸后EK的生物效應。研究目的:本研究通過對腸道微生物群的調節,研究EK的胃腸道毒性和Vek的解毒作用。雄性SD大鼠30只,隨機分為3組。連續7天口服0.5%CMC-Na(C組)、EK(甘遂組)或VEK(醋灸甘遂組)粉。結果:與EK相比,VEK中的10種有毒成分顯著降低。用醋炙后,VEK引起的副作用在組織病理學上、腸組織炎癥損傷指數、肝功能和氧化損傷指數減輕。EK的毒性可能與乳酸菌和Blautia屬密切相關。除此之外,醋灸后的EK增加了短鏈脂肪酸的產生,受到腸道微生物的調節。結論:醋炙后,主要益生菌比例增加,潛在致病菌減少。事實證明,用醋炙可以達到有效的解毒效果。

材料與方法

YANGBEN:GANSUIGENBU;JIANG100KEGANJINGDEGANSUIJINPAOZAI30KECUZHONG,ZAI260℃XIACHAOZHICHUXIANQINGWEIDEJIAOBAN。JIANGGANZAODEEKHEVEKMOCHENGFENMO,100MUGUOSHAI。

幸运飞艇官方开户 JIANCEPINGTAI:UPLC-QTOF/MS

SHIZHONGCANBIWUZHI:CONGEKGENZHONGFENLICHULAI,BINGTONGGUOGUANGPUFENXIJINXINGLEJIANDING。

DONGWUHEQUNTI:SDDASHUTIZHONG(230 ± 20) g,CHONGZUDESHUIHESHIWUSIYANGYIZHOUHOU,BEISUIJIFENWEILE3ZU,MEIZUSHIZHI,FENBIEGEIYUKOUFU0.5%CMC-Na、EKHUOVEKFENMO。

幸运飞艇官方开户 SHENGHUAFENXIHEZUZHIBINGLIXUEJIANCHA:

幸运飞艇官方开户 CAIJIJINGDONGMOXUEYANG,YONGYUCEDINGXUEQINGBINGANSUANZHUANANMEI(ALT)、TIANDONGANSUANZHUANANMEI(AST)、CHAOYANGHUAWUQIHUAMEI(SOD)、GUGUANGGANTAI(GSH)、BINGERQUAN(MDA)HERUSUANTUOQINGMEI(LDH)。

幸运飞艇官方开户 CAIJIDASHUXIAOCHANGZUZHI,JIANCEXIAOCHANGZHANMOSUNSHANGCHENGDU(FENWEI6GEDENGJI)HEMIANYIZUZHIHUAXUEYANJIU。

幸运飞艇官方开户 FENYANGCAIJIJI16S rDNAWEISHENGWUQUNLUOFENXI:

KUOZENG16 SrDNAJIYINV4-V5QU

515F:5‘-GTCCAGCCGCGG-3’;

907R:5‘-CCTCAATTCMTTGTTT-3’)。

DUANLIANZHIFANGSUANCEDING:GC-MS

結果展示

1.UPLC-QTOF/MSFENXIJIANDINGEKHEVEKZHIJIANQUBIEZUIDADECHENGFEN

幸运飞艇官方开户 WEILECHEDIPINGJIAEKHEVEKZHIJIANDEHUAXUEBIANHUA,DUILIANGGEYANGPINJINXINGUPLC-QTOF/MSFENXI,BINGLIYONGMassLynxRUANJIANDUIBAOLIUSHIJIAN(RT)、m/zLIZIDUIHELIZIQIANGDUDESHUJUJIJINXINGLEJINYIBUDECHULI。JIANDINGLE10GEZUFEN。QIZHONG,KansuinB,KansuinC,KansuinE,KansuinA,3-O-BENJIAXIAN-20-TUOYANGGUINUOCHUN,Kansuiphorin C,eupha-8,24-ERXI-3β,11β-ERCHUN-7-TONG,3-O-(2’E,4’Z-GUIERXIXIANJI)-20-O-YIXIANGUINUOCHUN,GANCANTONG,HE3-O-ERXI-1-(2’E,4’E-GUIERXIXIANJI)-20-O-YIXIANGUINUOCHUNBEISHIWEIDUXINGZUFEN,XIANGJIAOYUEK,VEKZHONGDEHANLIANGXIANZHUJIANGDI。

圖1 EK和VEK的UPLC-ESI+-MS總離子(TIC)色譜圖


幸运飞艇官方开户 2.DUIDASHUCHANGDAOZUZHIBINGLIXUEJIYANZHENGSUNSHANGZHIBIAODEYINGXIANG

幸运飞艇官方开户 JIYUH&ERANSE,CZUJUYOUWANZHENGSHANGPIXIBAOJIEGOUDEXIAOCHANGZUZHIDEZHENGCHANGZUZHIXUEXINGTAI(TU2A),ERZAIEKCZUDASHUZHONG,GUANCHADAOCHANGZHANMOZHONGDEXIANZHUSUNSHANG。EKCZUZHONGFASHENGDEDIANXINGZUZHIXUETEZHENGDETEZHENGZAIYUCHANGZHANMOGUYOUCENGDETUOLI,CHANGBIZHONGDEYANXINGXIBAOJINRUN(TU2B)。VEKCZUDASHUJUYOUXIANZHUGAISHANDECHANGZHANMOZUZHIXUEJIEGOU(TU2C)。EKCZUCHANGZHANMOSUNSHANGPINGFENXIANZHUGAOYUCZU,VEKCZUDIYUDUIZHAOZU(BIAO2)。

幸运飞艇官方开户 TU2. DI7TIANCHANGZUZHIDEDAIBIAOXINGZUZHIBINGLIXUETUXIANG。

幸运飞艇官方开户 ZHU:A,DUIZHAO(CZU);B,EK(680mg / Kg)CHULIDEDASHU(EKCZU);C,CUJIUEKFENMO680mg / Kg(VEKCZU)CHULIDEDASHU;YUANSHIFANGDABEISHUx100;HESEJIANTOUQUYUDAIBIAOYIGEYANZHONGDERONGMAOQUYUHEXIANTITUOLUO。

LIYONGKANGp65KANGTI,KANGICAM-1KANGTI,TONGGUOMIANYIZUZHIHUAXUEJISHUFAXIAN,YUCZUXIANGBI,EKCZUHEVEKCZUDASHUXIAOCHANGZUZHI(NF-κB)p65HEICAM-1BIAODAZENGJIA,SHANGPIXIBAOGAIZHANDANBAIBIAODAXIAJIANG。YUEKCZUXIANGBI,YONGVEKCZUDAOZHIp65HEICAM-1BIAODAXIANZHUJIANGDIBINGQIEE-GAIZHANDANBAIBIAODAZENGJIA(P <0.01)。JIEGUOXIANSHI,YONGCUJIUCHULIXIANZHUJIANGDILEp65HEICAM-1BIAODADEZENGJIA,BINGZENGJIALEEKYOUDAODEE-GAIZHANDANBAIDEJIANSHAO(TU3)。

幸运飞艇官方开户 3.DASHUGANGONGNENGJIXUEQINGYANGHUASUNSHANGZHIBIAODEBIANHUA

YUCZUXIANGBI,EKCZUHEVEKCZUDASHUXUEQINGAST(GUCAOZHUANANMEI)、ALT(GUBINGZHUANANMEI)、LDH(RUSUANTUOQINGMEI)HEMDA(BINGERQUAN)SHUIPINGXIANZHUSHENGGAO,GSH(GUGUANGGANTAIMEI)HESOD(CHAOYANGHUAWUQIHUAMEI)HUOXINGSHUIPINGXIANZHUJIANGDI。JIEGUOXIANSHI,CUJIUXIANZHUJIANGDILEEKYOUDAODEALTHEASTHUOXING。

TU4. EKHEVEKDUIZHENGCHANGDASHUXUEQINGGANGONGNENGHEYANGHUASUNSHANGZHIBIAODEYINGXIANG。

幸运飞艇官方开户 4.DASHUCHANGDAOWEISHENGWUQUNCEXUSHENDUYUαDUOYANGXINGDEBIJIAO

observed speciesQUXIANHERank-AbundanceQUXIANTUBIAOMING,MEIGEYANGBENDEWUZHONGFENBUJUNYUN,KEYONGYUDUOYANGXINGFENXI。ShannonZHISHUFANYINGLECELIANGWUZHONGDEJUNYUNDUHEFENGFUDU。EKCZUDEShannonZHISHUZHIXIANZHUGAOYUCHEEKCZU,VEKCZUDEShannonZHISHUZHIYUCZUXIANGSI。JIEGUOBIAOMING:EKDEDUXINGKENENGYUWEISHENGWUDUOYANGXINGDEZENGJIAYOUGUAN,VEKDEJIEDUZUOYONGKENENGYUWEISHENGWUDUOYANGXINGDEJIANSHAOYOUGUAN(TU5C)。

圖5. EK和VEK對大鼠腸道微生物α多樣性的影響。

5.DASHUCHANGDAOWEISHENGWUβDUOYANGXINGYUCHANGDAOJUNQUNFENGDUDEBIJIAO

幸运飞艇官方开户 ZAIGAIFENXIZHONG,LIYONGJIAQUANDEUniFracJULIZUOLEβDUOYANGXINGDEFENXI,KAOLVLEOTUZHIJIANDEXITONGFAYUJULI,KEYIZAIGETIZHONGXIDESHUIPINGHEGENGGAODEFENLEIXUESHUIPINGSHANGBUHUOCHAYI。PCATUXIANSHIZHESANZUCHULIKEMINGXIANQUFEN(TU6A)。LDAZAISANGEZUZHIJIANDEWEISHENGWUZUZHONGXIANSHICHUBUTONGDEFENLEIQUN(TU6B)。GAIFENLEITUBIAOMINGLEGEJIQUNTIZHONGBUTONGDEWEISHENGWUQUNLUOHUOWUZHONGJIEGOU。CladogramJIEGUOJINYIBUBIAOMINGSANZUZHIJIANDEBUTONGLEIBIEDEXIJUNFENBIEJULEI(TU6C)。

TU6. BIJIAOBUTONGZUZHIJIANDEFENBIANWEISHENGWUZU。

YONGEKHEEKCCHULIDEDASHU,CHANGDAOZHONGYOUDAIBIAOXINGDEYOUYIXIJUN(RUGANJUN)DEXIANGDUIFENGDUJIANGDI,ERCHANGCIJIXIANGGUANXIJUN(Blautia)DEXIANGDUIFENGDUZENGJIA。YUEKCZUXIANGBI,VEKCZUXIANSHIYISHENGJUNRUSUANGANJUNDEXIANGDUIFENGDUZENGJIA,BINGQIEBlautiaDEXIANGDUIFENGDUJIANGDI。ZHEXIEXIANGDUIFENGFUDEWEISHENGWUSHUDERETUYEXIANSHILIANGZHONGXIJUNZAISHUSHUIPINGSHANGSHIGAODUBIAODADEXIJUN。LIUWEIQIUJUNSHU(Ruminococcus)HENIPULEIWOJUN(Alloprevotella)DEXIANGDUIBAIFENBIEKCZUZHONGZUIDI,VEKCZUZHONGZUIGAO(TU8)。SHANGSHUJIEGUOBIAOMINGEKDEDUXINGHEVEKDEJIEDUKENENGYUSHANGSHUZHELIANGZHONGXIJUNSHUYOUGUAN。

幸运飞艇官方开户 TU7. ZAIMENHESHUDESHUIPINGSHANGDECHANGDAOWEISHENGWUQUNDEZUCHENG。

幸运飞艇官方开户 TU8. TONGGUO16S rDNAJIYINCEXUFENXIDEQIAN10GEXIJUNSHUDERETU。

6.EKHEVEKDUIDASHUFENBIANYANGPINZHONGDUANLIANZHIFANGSUANDEYINGXIANG

幸运飞艇官方开户 SANZUYANGPINZHONGDEZHUYAODEWUZHISHIYISUAN,BINGSUAN,DINGSUANHEWUSUAN,ZHEXIEWUZHITONGGUOJIANSHAOQUHUAYINZICHANSHENGHUOZHANFUFENZIBIAODALAIJIANSHAOCHANGDAOYANZHENG。YUCZUXIANGBI,EKCHEVEKCZUDUANLIANZHIFANGSUANDEXIANGDUINONGDUXIANZHUJIANGDI。VEKZUXIANGDUINONGDUXIANZHUZENGJIA。JIEGUOXIANSHI,CUJIUKENENGYUCHANGDAOWEISHENGWUQUNDIAOJIEDEDUANLIANZHIFANGSUANCHANLIANGXIANZHUZENGJIAYOUGUAN。

圖9. 通過GC-MS分析大鼠糞便樣品對SCFAs進行定量。

討論

     本研究通過給大鼠口服甘遂和醋灸后甘遂,得到的數據初步闡明了甘遂對腸道毒性作用以及通過醋灸后化學成分的改變和對動物的解毒作用。醋灸可以顯著減少甘的十種有毒成分,并且還可以顯著降低由組織病理學,肝功能,氧化損傷和炎癥損傷評估的肝和腸道毒性。甘遂經醋炙后,主要益生菌比例增加,潛在致病菌減少。事實證明,用醋炙可以達到有效的解毒效果。


原文鏈接:http://www.sciencedirect.com/science/article/pii/S0378874118304082