發(fā)布時間：2018-03-19 22:14

  本文選題：肝星狀細胞　切入點：第號染色體缺失的磷酸酶張力蛋白同源物基因　出處：《中國現(xiàn)代醫(yī)學雜志》2017年28期 　論文類型：期刊論文

【摘要】：目的探討野生型第10號染色體缺失的磷酸酶張力蛋白同源物基因(PTEN)過表達對體外活化肝星狀細胞(HSC)內(nèi)鈣離子濃度的影響。方法體外培養(yǎng)活化大鼠肝星狀細胞系(HSC-T6),利用腺病毒載體,將野生型PTEN基因轉染活化HSC;Western blot及實時熒光定量聚合酶鏈反應(q RT-PCR)檢測HSC的PTEN蛋白及m RNA表達;采用鈣離子熒光探針Rhod-2/AM,于激光掃描共聚焦顯微鏡下檢測HSC內(nèi)鈣離子濃度變化。實驗分組:(1)Control組:腺病毒轉染時以DMEM代替病毒液;(2)Ad-GFP組:轉染表達綠色熒光蛋白(GFP)的對照空病毒Ad-GFP;(3)Ad-PTEN組:轉染攜帶野生型PTEN基因并表達GFP的重組腺病毒Ad-PTEN。結果野生型PTEN在活化大鼠HSC大量表達,3組HSC內(nèi)鈣離子濃度比較,差異有統(tǒng)計學意義(P0.05),Ad-PTEN組HSC內(nèi)鈣離子濃度(251.60±90.88)低于Control組(1 953.95±132.99)及Ad-GFP組(1 937.57±115.17),而Control組與Ad-GFP組之間HSC內(nèi)鈣離子濃度比較,差異無統(tǒng)計學意義(P0.05)。結論野生型PTEN過表達可降低體外活化大鼠HSC內(nèi)鈣離子濃度。
[Abstract]:Objective to investigate the effect of overexpression of phosphatase tension protein congener (PTEN) on the concentration of calcium in activated hepatic stellate cells (HSCS) in vitro. Methods the activated rat hepatic stellate cell line HSC-T6 was cultured in vitro. Using adenovirus vectors, The expression of PTEN protein and m RNA in HSC were detected by transfection of wild-type PTEN gene into activated HSCC blot and real-time fluorescence quantitative polymerase chain reaction (Q RT-PCR). Calcium fluorescence probe Rhod-2 / AM was used to detect the change of calcium concentration in HSC under laser scanning confocal microscope. Ad-PTEN group: Ad-PTEN, a recombinant adenovirus carrying wild type PTEN gene and expressing GFP, was transfected into Ad-PTEN group. Results the Ca ~ (2 +) concentration of wild type PTEN in three groups of activated rat HSC was compared. There was a significant difference in the concentration of Ca ~ (2 +) in HSC between Ad-PTEN group and Control group (251.60 鹵90.88) and in Ad-GFP group (1 937.57 鹵115.17), respectively, and there was a significant difference between Control group and Ad-GFP group. Conclusion the overexpression of wild type PTEN can reduce the calcium concentration in HSC of rats activated in vitro.
【作者單位】： 華北理工大學附屬醫(yī)院消化內(nèi)科;
【基金】：河北省自然科學基金(No:H2013209327) 中國肝炎防治基金會天晴肝病研究基金資助項目(No:CFHPC20132078)
【分類號】：R575

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