中国中西医结合急救杂志
中國中西醫結閤急救雜誌
중국중서의결합급구잡지
INTEGRATED TRADITIONAL CHINESE AND WESTERN MEDICINE IN PRACTICE OF CRITICAL CARE MEDICINE
2013年
6期
369-373
,共5页
江远%张玲%邓远绮%何金洋
江遠%張玲%鄧遠綺%何金洋
강원%장령%산원기%하금양
肝星状细胞%肝纤维化%铁%去铁铵%活化%凋亡
肝星狀細胞%肝纖維化%鐵%去鐵銨%活化%凋亡
간성상세포%간섬유화%철%거철안%활화%조망
Hepatic stellate cell%Liver fibrosis%Iron%Desferrioxamine%Activation%Apoptosis
目的：探讨不同铁负载水平影响肝星状细胞（HSC）活化和凋亡的抗肝纤维化机制。方法根据细胞内不同铁负载水平将肝星状细胞株（HSC-T6细胞）分为空白对照组、铁沉积模型组、50μmol/L去铁铵组和25μmol/L去铁铵组共4组。用定量聚合酶链反应（PCR）检测HSC-T6细胞Ⅰ型胶原和转化生长因子-β1（TGF-β1）的mRNA表达；免疫组化法检测HSC-T6细胞α-平滑肌肌动蛋白（α-SMA）的表达；原位末端缺刻标记法（TUNEL）检测HSC-T6细胞的凋亡；电镜下观察HSC-T6细胞超微结构。结果与空白对照组比较，铁沉积模型组TGFβ1 mRNA表达虽有所增强，但差异无统计学意义（1.594±0.168比1.477±0.126，P＞0.05），Ⅰ型胶原mRNA表达则显著增强（1.354±0.076比1.197±0.104，P＜0.01）。50μmol/L和25μmol/L去铁铵均能下调Ⅰ型胶原和TGF-β1的mRNA表达，且50μmol/L去铁铵组优于25μmol/L去铁铵组（Ⅰ型胶原mRNA：0.391±0.076比0.688±0.060，TGF-β1 mRNA：0.421±0.068比0.714±0.090，均P＜0.01）。铁沉积可诱导HSC中α-SMA大量表达，却仅见有较少的凋亡细胞。而经去铁铵处理后HSC表达α-SMA明显减少，但HSC发生了凋亡。结论细胞内不同铁负载水平能够诱导HSC活化或凋亡，表明铁在调节HSC活化和凋亡过程中发挥重要作用，而且也揭示了铁螯合剂治疗肝纤维化的潜在作用。
目的：探討不同鐵負載水平影響肝星狀細胞（HSC）活化和凋亡的抗肝纖維化機製。方法根據細胞內不同鐵負載水平將肝星狀細胞株（HSC-T6細胞）分為空白對照組、鐵沉積模型組、50μmol/L去鐵銨組和25μmol/L去鐵銨組共4組。用定量聚閤酶鏈反應（PCR）檢測HSC-T6細胞Ⅰ型膠原和轉化生長因子-β1（TGF-β1）的mRNA錶達；免疫組化法檢測HSC-T6細胞α-平滑肌肌動蛋白（α-SMA）的錶達；原位末耑缺刻標記法（TUNEL）檢測HSC-T6細胞的凋亡；電鏡下觀察HSC-T6細胞超微結構。結果與空白對照組比較，鐵沉積模型組TGFβ1 mRNA錶達雖有所增彊，但差異無統計學意義（1.594±0.168比1.477±0.126，P＞0.05），Ⅰ型膠原mRNA錶達則顯著增彊（1.354±0.076比1.197±0.104，P＜0.01）。50μmol/L和25μmol/L去鐵銨均能下調Ⅰ型膠原和TGF-β1的mRNA錶達，且50μmol/L去鐵銨組優于25μmol/L去鐵銨組（Ⅰ型膠原mRNA：0.391±0.076比0.688±0.060，TGF-β1 mRNA：0.421±0.068比0.714±0.090，均P＜0.01）。鐵沉積可誘導HSC中α-SMA大量錶達，卻僅見有較少的凋亡細胞。而經去鐵銨處理後HSC錶達α-SMA明顯減少，但HSC髮生瞭凋亡。結論細胞內不同鐵負載水平能夠誘導HSC活化或凋亡，錶明鐵在調節HSC活化和凋亡過程中髮揮重要作用，而且也揭示瞭鐵螯閤劑治療肝纖維化的潛在作用。
목적：탐토불동철부재수평영향간성상세포（HSC）활화화조망적항간섬유화궤제。방법근거세포내불동철부재수평장간성상세포주（HSC-T6세포）분위공백대조조、철침적모형조、50μmol/L거철안조화25μmol/L거철안조공4조。용정량취합매련반응（PCR）검측HSC-T6세포Ⅰ형효원화전화생장인자-β1（TGF-β1）적mRNA표체；면역조화법검측HSC-T6세포α-평활기기동단백（α-SMA）적표체；원위말단결각표기법（TUNEL）검측HSC-T6세포적조망；전경하관찰HSC-T6세포초미결구。결과여공백대조조비교，철침적모형조TGFβ1 mRNA표체수유소증강，단차이무통계학의의（1.594±0.168비1.477±0.126，P＞0.05），Ⅰ형효원mRNA표체칙현저증강（1.354±0.076비1.197±0.104，P＜0.01）。50μmol/L화25μmol/L거철안균능하조Ⅰ형효원화TGF-β1적mRNA표체，차50μmol/L거철안조우우25μmol/L거철안조（Ⅰ형효원mRNA：0.391±0.076비0.688±0.060，TGF-β1 mRNA：0.421±0.068비0.714±0.090，균P＜0.01）。철침적가유도HSC중α-SMA대량표체，각부견유교소적조망세포。이경거철안처리후HSC표체α-SMA명현감소，단HSC발생료조망。결론세포내불동철부재수평능구유도HSC활화혹조망，표명철재조절HSC활화화조망과정중발휘중요작용，이차야게시료철오합제치료간섬유화적잠재작용。
Objective To investigate the anti-fibrosis mechanism from effects of difference in iron load levels on activation and apoptosis of hepatic stellate cells(HSCs). Methods According to the difference in iron load levels in HSCs,HSC-T6 cells were divided into four groups:blank control,iron deposition model,50μmol/L desferrioxamine and 25 μmol/L desferrioxamine groups. Quantitative polymerase chain reaction(PCR)was applied for the detection of collagen type Ⅰ and transforming growth factor-β1(TGF-β1)mRNA expressions of HSC-T6 cells. Immunohistochemical assay was used for the detection of α-smooth muscle actin(α-SMA)expression. The method of terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL)was used for the examination of apoptosis of HSC-T6 cells. Under electron microscope,the ultrastructures of HSC-T6 cells were observed. Results Compared with the blank control group,despite the TGF-β1 mRNA expression in iron deposition model group was increased,no statistical significant difference was seen(1.594±0.168 vs. 1.477±0.126, P>0.05),whereas collagen type I mRNA expression was significantly enhanced(1.354±0.076 vs. 1.197±0.104, P<0.01). Both 50μmol/L and 25μmol/L desferrioxamine could down-regulate collagen type I and TGF-β1 mRNA expressions,and the action of 50μmol/L desferrioxamine was superior to that of 25μmol/L desferrioxamine(collagen typeⅠmRNA:0.391±0.076 vs. 0.688±0.060,TGF-β1 mRNA:0.421±0.068 vs. 0.714±0.090,both P<0.01). Iron deposition could induce HSCs to expressα-SMA in great amount,while apoptosis could be seen scarcely in iron deposited HSCs. By desferrioxamine therapy,α-SMA expression of HSCs was decreased significantly,but some of the cells underwent apoptosis. Conclusion Different iron load levels inside HSCs can induce activation or apoptosis of the cells,showing that iron plays an important role in regulating the process of HSCs activation and apoptosis and revealing that desferrioxamine possesses the potential action for treatment of liver fibrosis.
