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《adma對(duì)肝星狀細(xì)胞活化及信號(hào)通路的實(shí)驗(yàn)研究》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫(kù)。
1、遵義醫(yī)學(xué)院碩士學(xué)位論文ADMA對(duì)肝星狀細(xì)胞活化及信號(hào)通路的實(shí)驗(yàn)研究姓名：趙娜申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：內(nèi)科學(xué)指導(dǎo)教師：劉春英201205遵義醫(yī)學(xué)院碩士學(xué)位論文中文摘要ADMA對(duì)肝星狀細(xì)胞活化及信號(hào)通路的實(shí)驗(yàn)研究中文摘要目的：觀察非對(duì)稱性二甲基精氨酸對(duì)肝星狀細(xì)胞增殖活化的影響及可能涉及的信號(hào)傳導(dǎo)通路。方法：體外培養(yǎng)HSC．T6細(xì)胞，取對(duì)數(shù)生長(zhǎng)期細(xì)胞進(jìn)行實(shí)驗(yàn)，分組培養(yǎng)。予不同濃度的ADMA及P38MAPK阻斷劑(SB203580)處理HSC，應(yīng)用倒置相差顯微鏡觀察細(xì)胞形態(tài)變化；通過(guò)MTT法檢測(cè)藥物對(duì)細(xì)胞增殖的影響；應(yīng)用RT—PCR法檢測(cè)藥物作用后P3
2、8MAPK、TGF．Bl、I型膠原mRNA表達(dá)水平的變化；通過(guò)ELISA法檢測(cè)各實(shí)驗(yàn)組I型膠原蛋白的表達(dá)變化。結(jié)果：1、通過(guò)鏡下觀察細(xì)胞形態(tài)變化顯示：隨著ADMA濃度的增加，細(xì)胞體積逐漸增大，較對(duì)照組伸展更明顯，生長(zhǎng)狀態(tài)良好。2、MTT實(shí)驗(yàn)表明：ADMA呈濃度一時(shí)間依賴性促進(jìn)HSC增殖，在ADMA(10“mol／1)、作用48h時(shí)細(xì)胞增殖率最高；SB203580可抑制HSC增殖，隨著SB203580濃度增加，HSC增殖抑制率逐漸增加。3、RT—PCR實(shí)驗(yàn)結(jié)果顯示：ADMA呈濃度依賴性上調(diào)P38MAPK、TGF．pl、I型膠原mRNA表達(dá)；而SB
3、203580則使P38MAPK、TGF．Bl、I型膠原mRNA表達(dá)水平下降。4、ELISA實(shí)驗(yàn)表明隨著ADMA濃度增加，I型膠原蛋白表達(dá)逐漸增多；SB203580則呈濃度依賴性降低I型膠原蛋白表達(dá)。結(jié)論：ADMA促進(jìn)HSC增殖活化進(jìn)一步促進(jìn)ECM的增多，可能與激活P38MAPK信號(hào)通路，上調(diào)TGF．pl表達(dá)，增加膠原合成有關(guān)；SB203580能抑NHSC增殖、P38MAPK信號(hào)傳導(dǎo)通路及TGF．Bl表達(dá)，減少膠原合成。關(guān)鍵詞：非對(duì)稱性二甲基精氨酸：P38MAPK阻斷劑(SB203580)；肝星狀細(xì)胞；活化；信號(hào)通路遵義醫(yī)學(xué)院碩士學(xué)位論文英文摘要
4、StudyoftheeffectofADMAOnhepaticstellatecellsproliferationanditspossiblesignalpathway．AbstractObjective：ToinvestigatetheeffectsofADMA(asymmetricdimethylarginine；theendogenousinhibitorofNOsynthesis)onhepaticstellatecells(HSC)proliferationanditspossiblesignalpathway．Methods：Aft
5、erculturedHSC—T6dellswereexposedtodifferentconcentrationsofADMAandSB203580．Invertedphasecontrastmicroscopewasusedtoobservecellsmorphologicchange；cellproliferationwasdeterminedbymethylthiazolyltetrazolium(MTT)assay，andreal—timePCRwasusedtodetectthegeneexpressionsofP38MAPK、TGF
6、-131、type·Icollagens；type—IcollagenproteinchangeswasmeasuredbyELISA．Results：Undertheinvertedphasecontrastmicroscope，wecouldobservethatcellvolumeincreasegradually，stretchmoreapparemthanthecontrolgroup，growthisingoodcondition．withtheinfluenceofdiffe：rentconcentrationsofADMA．MT
7、TshowedADMAcanprompttheproliferationofhepaticstellatecellinatimeandconcentrationdepenence，inthecontrast，SB203580inhibittheproliferationofHSC．PCRandELISAindicatedthatADMAcouldinducetheincreaseofP38MAPK、TGF-p1、type·IcollagensrnRNAandproteinproductioninaconcentration．dependentm
8、anner；while，SB30．2580couldsignificantlyinhibittheexpressionofP38MAPK、TGF-pl
