[1]李冬,戴立里,余斌斌,等.丹参素对大鼠肝星状细胞增殖、凋亡及NF-κB活性的影响[J].第三军医大学学报,2009,31(08):724-728.
 LI Dong,DAI Li-li,YU Bin-bin,et al.Tanshinol induces proliferation, apoptosis and NF-κB activation in cultured rat hepatic stellate cells after IL-1β inducement[J].J Third Mil Med Univ,2009,31(08):724-728.
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丹参素对大鼠肝星状细胞增殖、凋亡及NF-κB活性的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第08期
页码:
724-728
栏目:
论著
出版日期:
2009-04-30

文章信息/Info

Title:
Tanshinol induces proliferation, apoptosis and NF-κB activation in cultured rat hepatic stellate cells after IL-1β inducement
作者:
李冬戴立里余斌斌姜中华
重庆医科大学第二附属医院消化内科
Author(s):
LI Dong DAI Li-li YU Bin-bin JIANG Zhong-hua
Department of Gastroenterology, Second Affiliated Hospital, Chongqing Medical University, Chongqing 400010, China
关键词:
肝纤维化丹参素肝星状细胞增殖凋亡NF-κBIL-1β
Keywords:
hepatic fibrosis tanshinol hepatic stellate cells proliferation apoptosis NF-κB IL-1β
分类号:
R285.5;R322.47;R329.28
文献标志码:
A
摘要:
目的    观察丹参素对IL-1β刺激的肝星状细胞增殖、凋亡及NF-κB活性的影响,探讨丹参素抗肝纤维化的分子机制。    方法    用原位-密度梯度离心法提取肝星状细胞(hepatic stellate cell, HSCs),以不同浓度的丹参素作用于IL-1β刺激的HSCs,MTT法、AO/EB免疫荧光和Annexin V-FITC/PI分别检测HSCs的增殖和凋亡;免疫细胞化学、ELISA法、Western blot分别检测Ⅲ型胶原的生成、NF-κB 核转位及细胞质p-IκBα、细胞核NF-κB p65的表达。    结果    与对照组相比,IL-1β刺激组HSCs增殖明显增加(P<0.01),凋亡率明显减低(P<0.05),Ⅲ型胶原的合成和分泌明显增加(P<0.01);不同浓度丹参素作用24 h后HSCs增殖明显减低(P<0.01),凋亡明显增加(P<0.01),以早期凋亡为主,Ⅲ型胶原的合成和分泌亦明显减少(P<0.05,P<0.01);Western blot结果显示IL-1β作用30 min时细胞质p-IκBα、细胞核NF-κB p65蛋白表达明显增加(P<0.01),出现核转位;免疫细胞化学显示细胞核内p65阳性染色细胞增多浓染,提示IL-1β可以诱导HSCs的NF-κB活性;丹参素组细胞质p-IκBα、细胞核NF-κB p65蛋白表达明显减低(P<0.05,P<0.01),核转位减少,细胞核内p65阳性染色细胞数减少淡染,提示丹参素可以抑制IL-1β诱导的HSCs NF-κB活性。    结论    丹参素抑制IL-1β诱导的HSCs增殖,促使HSCs凋亡,减少Ⅲ型胶原的合成和分泌,其机制可能与抑制NF-κB活性有关。
Abstract:
Objective    To investigate the effects of tanshinol on the proliferation, apoptosis and NF-κB activation in rat hepatic stellate cells (HSCs) after IL-1β inducement, and to elucidate the anti-fibrotic molecular mechanisms of tanshinol.     Methods    The rat HSCs was isolated with collagenase in situ liver recirculation perfusion and cultured in vitro. The cells were divided into 5 groups: normal control, IL-1β treatment group (10 ng/ml), and tanshinol group 1, 2 and 3. The later 3 groups were pretreated with tanshinol at the concentrations of 0.062 5, 0.125 and 0.25 mmol/L respectively followed by 10 ng/ml IL-1β treatment 1 h later. MTT colorimetric assay was used to detect the proliferation of HSCs. AO/EB immunoflurorescence microscopy and combination Annexin-V-FITC/PI double-labelimmunofluorescence with flow cytometer were employed to examine the apoptosis of HSCs. Synthesis and secretion of collagen Ⅲ were detected by the quantitative immunocytochemical assay and ELISA respectively. The amounts of cytoplasm p-IκBα and NF-κB p65, and nuclear NF-κB p65 in HSCs were determined by Western blotting. Immunocytochemical staining and Western blotting were used to observe nuclear translocation of NF-κB p65.     Results    IL-1β increased the proliferation of HSCs (P<0.01), reduced the cell apoptosis (P<0.05), and facilitated the synthesis and secretion of collagen Ⅲ in HSCs (P<0.05, P<0.01) in comparison with normal control. After treated with different concentration of tanshinol, HSC’s proliferation was prominently reduced (P<0.01) and its apoptosis was increased (P<0.01), especially for early apoptosis. Tanshinol also degraded the synthesis and secretion of collagen Ⅲ in HSCs, especially in 0.25 mmol/L concentration group (P<0.01). Western blotting showed that IL-1β elevated amounts of cytoplasm p-IκBa and nuclear NF-κB p65 (P<0.01), decreased amounts of cytoplasm p65 markedly (P<0.05), then facilitated nuclear translocation of p65. Immunocytochemical staining indicated that the number of nuclear NF-κB p65 positive HSCs were increased and the color was deepened. IL-1β induced NF-κB activation in rat HSCs. Tanshinol reduced the amounts of cytoplasm p-IκBa and nuclear NF-κB p65 (P<0.05, P<0.01), and decreased nuclear translocation of p65 markedly. Immunocytochemical staining showed that the number of nuclear NF-κB p65 positive HSCs was decreased and the color grew pallid. Above results indicated that tanshinol inhibited IL-1β-induced NF-κB activation in rat HSCs.     Conclusion    Tanshinol inhibits the proliferation of HSCs, facilitates the cell apoptosis, and reduces the synthesis and secretion of collagen   Ⅲ  through inhibiting NF-κB activation via suppressing hepatic fibrosis.

参考文献/References:

李冬,戴立里,余斌斌,等. 丹参素对大鼠肝星状细胞增殖、凋亡及NF-κB活性的影响[J]. 第三军医大学学报,2009,31(8):724-728.

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更新日期/Last Update: 2009-04-15