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1、1450重慶醫(yī)學(xué)2017年4月第46卷第11期論著·基礎(chǔ)研究doi:10.3969/j.issn.1671-8348.2017.11.003*辣椒素對(duì)體外肝細(xì)胞脂質(zhì)代謝的影響研究孫玉1,劉佳2#,寧波1,楊朝霞1△(1.重慶醫(yī)科大學(xué)附屬第二醫(yī)院消化內(nèi)科400010;2.湖北省武漢市普仁醫(yī)院消化與肝病科430081)[摘要]目的探討辣椒素對(duì)脂肪變肝細(xì)胞內(nèi)脂質(zhì)沉積的影響及自噬表達(dá)情況,為脂肪性肝病防治的研究提供新的思路和靶點(diǎn)。方法用油酸(40μg/mL)誘導(dǎo)肝原代細(xì)胞LO2細(xì)胞株建立非酒精性脂肪性肝病(NAFLD)細(xì)胞模型。實(shí)驗(yàn)分為空白對(duì)照組、
2、油酸組、辣椒素組[油酸+辣椒素(100μmol/L)]。油紅O染色和三酰甘油試劑盒檢測(cè)肝細(xì)胞內(nèi)脂肪變程度;Westernblot檢測(cè)自噬體標(biāo)記分子p62和LC3的表達(dá)水平并計(jì)算LC3-Ⅱ/LC3-Ⅰ比值。結(jié)果40μg/mL油酸處理LO2細(xì)胞24h后,油紅O染色觀察,油酸組肝細(xì)胞中可見大小不等的橘紅色脂滴,而空白對(duì)照組無明顯橘紅色脂滴形成,油酸在體外成功建立NAFLD模型。與油酸組相比,辣椒素組肝細(xì)胞內(nèi)三酰甘油水平顯著降低(P<0.05),脂滴明顯減少,p62蛋白水平明顯降低(P<0.05),而LC3-Ⅱ/LC3-Ⅰ比值明顯上調(diào)(P<0.05
3、)。結(jié)論在體外利用油酸誘導(dǎo)LO2細(xì)胞株建立NAFLD模型,辣椒素刺激NAFLD細(xì)胞,上調(diào)NAFLD細(xì)胞自噬水平,減少細(xì)胞內(nèi)脂質(zhì)沉積,但具體機(jī)制還需進(jìn)一步研究。[關(guān)鍵詞]油酸;辣椒辣素;非酒精性脂肪性肝病[中圖分類號(hào)]R575.5[文獻(xiàn)標(biāo)識(shí)碼]A[文章編號(hào)]1671-8348(2017)11-1450-04Effectsofcapsaicinonlipidmetabolismoflivercellsinvitro*1,LiuJia2#,NingBo1,YangZhaoxia1△SunYu(1.DepartmentofGastroenterol
4、ogy,theSecondAffiliatedHospitalofChongqingMedicalUniversity,Chongqing,400010,China;2.DepartmentofGastroenterologyandHepatic,PurenHospitalofWuhanCity,Wuhan430081,China)[Abstract]ObjectiveToobservetheeffectofcapsaicinonlipiddepositioninlivercellsandtheexpressionofautophagy,t
5、oprovidenewideasandtargetsforthestudyoffattyliverdisease.MethodsThemodelofnonalcoholicfattyliverdisease(NAFLD)wasestablishedbyinducingLGFcelllineofliverprimarycellswitholeicacid(40μg/mL).Theexperimentwasdividedintoblankcontrolgroup,oleicacidgroup,capsaicingroup[oleicacid+c
6、apsaicin(100μmol/L)].ThelevelofintracellularlipidwasdetectedbyoilredOstainingandtriglyceride(TG)kit.Westernboltwasusedtodetecttheexpressionlevelofautophagymarkermoleculep62andLC3,andtocalculatetheratioofLC3-Ⅱ/LC3-Ⅰ.ResultsAftertreatedwith40μg/mLoleicacidfor24h,theoilredOst
7、ai-ningshowedthatorange-redlipiddropletswerefoundintheoleicacidgroup,buttherewasnoobviousorange-redlipiddropletfor-mationintheblankgroup,whichsuggestingthattheNAFLDcellmodelwasestablishedsuccessfullybyusing40μg/mLoleicacidinvitro.OilredOstainingwasobserved,thecellsinthecap
8、saicingroupweresignificantlylessthantheoleicacidgroup,andthecontentoftriglycerideintheliv