中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (46): 8706-8711.doi: 10.3969/j.issn.1673-8225.2011.46.037

• 组织构建与中医药 tissue construction and traditional Chinese medicine • 上一篇    下一篇

川芎嗪可影响肿瘤坏死因子α刺激肝星状细胞结缔组织生长因子、核因子κB及相关基因产物的表达

李艳芳，李孝生   

  1. 重庆医科大学附属第二医院消化内科，重庆市  400010
  • 收稿日期:2011-04-01 修回日期:2011-07-11 出版日期:2011-11-12 发布日期:2011-11-12
  • 通讯作者: 李孝生，硕士，教授，硕士生导师，重庆医科大学附属第二医院消化内科，重庆市 400010 LXS6896@126. com
  • 作者简介:李艳芳★，女，1985年生，山东省聊城市人，重庆医科大学在读硕士，主要从事肝纤维化的防治研究。 liyanfang3715183@163.com
  • 基金资助:

    重庆市科委自然科学基金资助项目(2006BB5428)，课题名称：川芎嗪对HSC的影响及CTGF信号转导的调控机制探讨。

Effects of tetramethylpyrazine on expression of connective tissue growth factor, nuclear factor kappa B and its related gene products in tumor necrosis factor-alpha stimulated hepatic stellate cells

Li Yan-fang, Li Xiao-sheng   

  1. Department of Gastroenterology, Second Affiliated Hospital of Chongqing Medical University, Chongqing  400010, China
  • Received:2011-04-01 Revised:2011-07-11 Online:2011-11-12 Published:2011-11-12
  • Contact: Li Xiao-sheng, Master, Professor, Master’s supervisor, Department of Gastroenterology, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China LXS6896@126.com
  • About author:Li Yan-fang★, Studying for master's degree, Department of Gastroenterology, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China liyanfang3715183@ 163.com
  • Supported by:

    the Natural Science Foundation of Committee of Science and Technology of Chongqing, No. 2006BB5428*

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摘要：

背景：川芎嗪延缓肝纤维化形成的机制尚不清楚。
目的：观察川芎嗪对肿瘤坏死因子α刺激的肝星状细胞增殖及结缔组织生长因子、核因子κB及其相关基因产物白细胞介素6表达的影响。
方法：体外培养HSC-T6细胞株，取对数生长期的细胞用于实验。实验分为4组：对照组仅加入细胞；TNF-α组加入10 μg/L TNF-α；川芎嗪干预组先加入终浓度为10 μg/L的TNF-α作用30 min后，分别加入川芎嗪50，100，200，400，600， 1 000 mg/L；PDTC组先加入终浓度为10 μg/L的TNF-α作用30 min后，再加入终浓度18 μmol/L的核因子κB阻断剂PDTC。
结果与结论：MTT结果显示100，200，400，600，1 000 mg/L的川芎嗪均能抑制HSC-T6增殖，且呈剂量依赖性。免疫细胞化学染色及Western blot检测发现10 μg/L的肿瘤坏死因子α刺激后，HSC-T6细胞结缔组织生长因子、核因子κB及白细胞介素6的表达明显增多(P < 0.01或P < 0.05)，200，400，600 mg/L的川芎嗪及18 μmol/L的PDTC均可明显降低肿瘤坏死因子α刺激后HSC-T6细胞结缔组织生长因子、核因子κB及白细胞介素6的表达(P < 0.01)，且随着川芎嗪质量浓度的增加，抑制作用增强，PDTC的抑制作用最明显。相关性分析结果显示HSC-T6细胞结缔组织生长因子和核因子κB的表达呈正相关(r=0.980，P < 0.01)。说明川芎嗪可以抑制肝星状细胞结缔组织生长因子、核因子κB及白细胞介素6的表达，抑制肝星状细胞的增殖。

关键词: 川芎嗪, 肿瘤坏死因子&alpha, 肝星状细胞, 结缔组织生长因子, 核因子&kappa, B

Abstract:

BACKGROUND: Previous studies have shown that tetramethylpyrazine (TMP) can decrease expression of connective tissue growth factor (CTGF), nuclear factor kappa B (NF-κB) and delay the formation of hepatic fibrosis. But the mechanism and the correlation of CTGF and NF-κB was not clear.           
OBJECTIVE: To investigate the effects of TMP on expression of CTGF, NF-κB and its related gene product interleukin-6 (IL-6) in tumor necrosis factor-alpha (TNF-α) stimulated hepatic stellate cells.
METHODS: Hepatic stellate cell-T6 (HSC-T6) was cultured to logarithmic growth phase for experiments. Cells were divided into four groups: control group: only cells were added; TNF-α group: 10 μg/L TNF-α was added; TMP intervention group: pretreated with 10 μg/L TNF-α for 30 minutes and then 50,100, 200, 400, 600, 1 000 mg/L TMP was added; PDTC group: pretreated with   10 μg/L TNF-α for 30 minutes, and then stimulated with 18 μmol/L PDTC, a blocker of NF-κB.
RESULTS AND CONCLUSIONS: MTT assay results showed that TMP inhibited the proliferation of HSC-T6 at a concentration of 100, 200, 400, 600, and 1 000 mg/L in a concentration dependent manner (P < 0.05). Immunocytochemical staining and western blot analysis showed that 200, 400, 600 mg/L TMP and 18 μmol/L PDTC could decrease the expression of TNF-α-induced CTGF, NF-κB and IL-6 (P < 0.01) in HSC-T6 cells and with increasing concentrations of TMP, the inhibitory effects were enhanced, and PDTC could inhibit their expression effectively. Correlation analysis showed that the expression of cTGF and NF-κB in HSC-T6 cells was positively correlated (r=0.980, P < 0.01).These data indicated that TMP could inhibit the expression of cTGF, NF-κB and IL-6 in hepatic stellate cells and inhibit the proliferation of hepatic stellate cells.

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