中华普通外科杂志
中華普通外科雜誌
중화보통외과잡지
CHINESE JOURNAL OF GENERAL SURGERY
2012年
10期
821-824
,共4页
俞富祥%符竣惠%谢少华%张启瑜
俞富祥%符竣惠%謝少華%張啟瑜
유부상%부준혜%사소화%장계유
胆红素%肝硬化%肝星状细胞%活化
膽紅素%肝硬化%肝星狀細胞%活化
담홍소%간경화%간성상세포%활화
Bilirubin%Liver cirrhosis%Hepatic stellate cells%Activation
目的 探讨低浓度胆红素( bilirubin)对活化态肝星状细胞(hepatic stellate cells,HSCs)增殖、活化、凋亡的影响.方法 从肝脏分离纯化培养HSCs,DCFH-DA法检测不同浓度胆红素对HSCs中活性氧的影响.通过CCK-8比色法检测胆红素对HSCs增殖的影响.Western blot检测HSCsα-肌动蛋白(d-SMA)的表达.流式细胞仪检测胆红素对HSCs凋亡的影响.PCR检测胆红素对HSCs纤维化相关基因表达的影响.结果 分离并培养HSCs,低浓度胆红素浓度(0、1、10、20 mg/L)抑制HSCs中活性氧的产生,明显抑制HSCs的增殖(0.624±0.092,0.536±0.127,0.407±0.033,0.399±0.022,F=13.454,P＜0.05)及α-SMA的表达(339 ±2,336±10,246±7,242±5,3.7±0.3,F=191.107,P＜0.05),促进HSCs的凋亡(2.69±0.07％,2.95±0.10％,4.41±0.22％,4.91±0.86％,F=34.731,P＜0.05),改变HSCs纤维化相关基因的表达,TIMP-1 mRNA/MMP-2 mRNA的比值呈降低趋势(54±2,65±2,47 ±2,44±2,F=73.400,P＜0.05).结论 低浓度胆红素具有抑制HSCs活化增殖,促进凋亡的潜能.胆红素的作用可能与其抗氧化功能有关.
目的 探討低濃度膽紅素( bilirubin)對活化態肝星狀細胞(hepatic stellate cells,HSCs)增殖、活化、凋亡的影響.方法 從肝髒分離純化培養HSCs,DCFH-DA法檢測不同濃度膽紅素對HSCs中活性氧的影響.通過CCK-8比色法檢測膽紅素對HSCs增殖的影響.Western blot檢測HSCsα-肌動蛋白(d-SMA)的錶達.流式細胞儀檢測膽紅素對HSCs凋亡的影響.PCR檢測膽紅素對HSCs纖維化相關基因錶達的影響.結果 分離併培養HSCs,低濃度膽紅素濃度(0、1、10、20 mg/L)抑製HSCs中活性氧的產生,明顯抑製HSCs的增殖(0.624±0.092,0.536±0.127,0.407±0.033,0.399±0.022,F=13.454,P＜0.05)及α-SMA的錶達(339 ±2,336±10,246±7,242±5,3.7±0.3,F=191.107,P＜0.05),促進HSCs的凋亡(2.69±0.07％,2.95±0.10％,4.41±0.22％,4.91±0.86％,F=34.731,P＜0.05),改變HSCs纖維化相關基因的錶達,TIMP-1 mRNA/MMP-2 mRNA的比值呈降低趨勢(54±2,65±2,47 ±2,44±2,F=73.400,P＜0.05).結論 低濃度膽紅素具有抑製HSCs活化增殖,促進凋亡的潛能.膽紅素的作用可能與其抗氧化功能有關.
목적 탐토저농도담홍소( bilirubin)대활화태간성상세포(hepatic stellate cells,HSCs)증식、활화、조망적영향.방법 종간장분리순화배양HSCs,DCFH-DA법검측불동농도담홍소대HSCs중활성양적영향.통과CCK-8비색법검측담홍소대HSCs증식적영향.Western blot검측HSCsα-기동단백(d-SMA)적표체.류식세포의검측담홍소대HSCs조망적영향.PCR검측담홍소대HSCs섬유화상관기인표체적영향.결과 분리병배양HSCs,저농도담홍소농도(0、1、10、20 mg/L)억제HSCs중활성양적산생,명현억제HSCs적증식(0.624±0.092,0.536±0.127,0.407±0.033,0.399±0.022,F=13.454,P＜0.05)급α-SMA적표체(339 ±2,336±10,246±7,242±5,3.7±0.3,F=191.107,P＜0.05),촉진HSCs적조망(2.69±0.07％,2.95±0.10％,4.41±0.22％,4.91±0.86％,F=34.731,P＜0.05),개변HSCs섬유화상관기인적표체,TIMP-1 mRNA/MMP-2 mRNA적비치정강저추세(54±2,65±2,47 ±2,44±2,F=73.400,P＜0.05).결론 저농도담홍소구유억제HSCs활화증식,촉진조망적잠능.담홍소적작용가능여기항양화공능유관.
Objective To investigate the influence of low bilirubin level on hepatic stellate cells (HSCs) in vitro. Methods HSCs were isolated and cultured from the liver of SD rats. The effect of bilirubin in different concentration on reactive oxygen in HSCs was determined by DCFH-DA kit. The proliferation of HSCs was tested by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of HSCs was tested by Western blot.The apoptosis of HSCs was tested by flow cytometry.The fibrosis-related genes were tested by PCR. Results HSCs were isolated and cultured successfully.Bilirubin in low concentration (0,1,10,20 mg/L) inhibited the generation of the reactive oxygen.Proliferation and α-SMA expression of HSCs was inhibited by bilirubin (0.624 ± 0.092,0.536 ± 0.127,0.407 ± 0.033,0.399 ± 0.022,F =13.454,P ＜0.05 ; 339 ± 2,336 ± 10,246 ± 7,242 ± 5,3.7 ± 0.3,F =191.107,P ＜ 0.05 ) and the apoptosis of HSCs was promoted by bilirubin(2.69 ±0.07％,2.95 ±0.10％,4.41 ±0.22％,4.91 ±0.86％,F =34.731,P ＜0.05 ).Bilirubin in low concentration changed the expression of fibrosis-related genes in HSCs.The ratio of TIMP-1mRNA/MMP-2mRNA decreased (54 ± 2,65 ± 2,47 ± 2,44 ± 2,F =73.400,P ＜ 0.05).Conclusions Bilirubin in low concentration inhibits the proliferation and activation of hepatic stellate cells,orobablv bv a mechanism in which bilirubin promoted antioxidative function.
