摘要
目的 了解槲皮素对肝脏星状细胞 (HSCs)通过转化生长因子 (TGF)β1诱导的结缔组织生长因子(CTGF)及纤维连接素 (FN)表达的影响。方法 通过胶原酶与链霉蛋白酶原位灌注、Nycondenz一步法梯度离心分离雄性 Wistar大鼠 HSCs。培养活化的 HSCs无血清饥饿后与 10 - 9～ 10 - 5m ol/ L浓度的槲皮素孵育 2 4 ,4 8或 72h。以流式细胞术测定 TGFβ1的表达与分泌。逆转录聚合酶链反应 (RT- PCR)检测 CTGF基因表达 ,免疫组化检测FN表达。结果 流式细胞术检测免疫荧光强度显示 10 - 8～ 10 - 5m ol/ L浓度槲皮素可抑制 HSCs TGFβ1表达 ;10 - 7mol/ L浓度槲皮素孵育 4 8h可显著抑制 HSCs TGFβ1表达 (特异性平均荧光强度分别为 13.33± 2 .4 4和 18.0 8±2 .5 4 ,t=16 .5 2 ,P<0 .0 1)。 TGFβ1可明显促进活化 HSCs CTGF m RNA表达 ,但 10 - 7mol/ L浓度槲皮素在 72 h内几乎可完全拮抗此效应。与对照相比 ,10 - 7mol/ L浓度槲皮素孵育 4 8h可显著抑制 HSCs FN表达。结论 槲皮素可抑制 TGFβ1信号通路 ,包括抑制 TGFβ1、FN表达及 CTGF的基因表达 ,具有潜在的抗肝纤维化治疗作用。
Objective To investigate the effect of quercetin on the expression of connective tissue growth factor (CTGF) and fibronectin(FN) in hepatic stellate cells (HSCs) stimulated with transforming growth factor(TGF)β1. Methods HSCs of rat were isolated by in situ perfusion and gradients centrifugation; the activated HSCs were incubated with quercetin (10 -9 10 -5mol/L) for 24, 48 and 72 h. TGFβ1 was detected by flow cytometry. The expression of CTGF was detected by reverse transcription-polymerase chain reaction (RT-PCR), and then FN was detected by immunohistochemistry. Results Quercetin (10 -8 to 10 -5 mol/L) inhibited the expression of TGFβ1 in HSCs. TGFβ1 expression decreased after the HSCs being incubated with quercetin (10 -7 mol/L) for 48 h (mean fluorescence: 13.33±2.44 and 18.08±2.54,t=16.52, P<0.01). TGFβ1 increased the expression of CTGF mRNA in HSCs, but this effect was abrogated by quercetin (10 -7 mol/L) within 72 h. Quercetin(10 -7 mol/L) significantly inhibited the expression of FN in HSCs. Conclusion Quercetin may have an inhibitory effect on the signal pathways of TGFβ1, including the expression of TGFβ1, FN and CTGF.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2004年第6期802-805,共4页
Journal of Sichuan University(Medical Sciences)
