[1]陈璐,何文华,朱萱,等.熊果酸对肝星状细胞NADPH 氧化酶-Hedgehog 信号通路的影响[J].陆军军医大学学报(原第三军医大学学报),2014,36(05):427-431.
 Chen Lu,He Wenhua,Zhu Xuan,et al.Ursolic acid suppressed NADPH oxidase-Hedgehog signaling pathway in hepatic stellate cells in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(05):427-431.
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熊果酸对肝星状细胞NADPH 氧化酶-Hedgehog 信号通路的影响(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
36卷
期数:
2014年第05期
页码:
427-431
栏目:
论著
出版日期:
2014-03-15

文章信息/Info

Title:
Ursolic acid suppressed NADPH oxidase-Hedgehog signaling pathway in hepatic stellate cells in vitro
作者:
陈璐何文华朱萱李弼民张焜和施凤张新华
南昌大学第一附属医院消化内科, 江西省消化疾病研究所
Author(s):
Chen Lu He Wenhua Zhu Xuan Li Bimin Zhang Kunhe Shi Feng Zhang Xinhua
Department of Gastroenterology, First Affiliated Hospital, Nanchang University, Jiangxi Provincial Institute of Gastroenterology, Nanchang, Jiangxi Province, 330006, China
关键词:
熊果酸肝星状细胞Hedgehog信号通路NADPH氧化酶Rac1
Keywords:
ursolic acid hepatic stellate cells Hedgehog signal pathway NADPH oxidase Rac1
分类号:
R285.5; R322.47; R329.26
文献标志码:
A
摘要:
目的      探讨熊果酸(ursolic acid,UA)对大鼠肝星状细胞(HSC-T6)的NADPH氧化酶(NOX)-Hedgehog(Hh)信号通路的影响。      方法        将处于对数生长期的HSC-T6细胞分为6组:正常对照组、瘦素组(100 ng/mL)、熊果酸自身对照组(50 μmol/L)、DPI自身对照组(20 μmol/L)、熊果酸干预组(瘦素+熊果酸)、DPI干预组(瘦素+DPI)。在药物作用12 h后,提取总RNA,采用RT-PCR法检测Shh、Smo、Gli1/2的表达;药物作用HSC-T6 细胞24 h后,提取总蛋白,采用Western blot分别检测Rac1和Gli2的表达;药物作用12、24、48 h后,采用MTT法检测HSC-T6细胞的增殖情况。      结果      RT-PCR分析显示,瘦素刺激HSC-T6细胞12 h后Smo mRNA表达较正常对照组升高(P<0.05);Shh、Gli2 mRNA表达稍高于正常对照组,但差异无统计学意义(P>0.05); 熊果酸干预后Shh、Smo和Gli2 mRNA表达明显低于瘦素组及正常对照组(均P<0.05);瘦素对HSC-T6细胞Gli1 mRNA的表达无影响,而熊果酸及DPI干预也不影响HSC-T6细胞Gli1 mRNA 的表达。瘦素刺激HSC-T6细胞24 h后,Rac1和 Gli2蛋白的表达较正常对照组升高(P<0.01);熊果酸干预后Rac1和Gli2蛋白的表达明显低于瘦素组(P<0.01)。MTT分析显示瘦素促进HSC-T6细胞增殖,熊果酸干预12 h后HSC-T6细胞的生长抑制率均显著高于瘦素组(P<0.01),随着作用时间延长,熊果酸对细胞生长的抑制作用进一步增强。      结论        熊果酸能抑制瘦素诱导的HSC-T6细胞Hh信号通路Shh、Smo、Gli2 mRNA和 Gli2蛋白表达。熊果酸通过抑制NOX亚基Rac1蛋白表达进而抑制Hh信号通路可能是它抑制肝星状细胞生长增殖的机制之一。
Abstract:
Objective        To determine the effect of ursolic acid (UA) on NADPH oxidase (NOX)- Hedgehog (Hh) signaling pathway in hepatic stellate cells(HSCs).       Methods        Culture-activated HSC-T6 cells were divided into 6 groups: normal control group, leptin group (100 ng/mL), UA group (50 μmol/L), NOX inhibitor DPI (20 μmol/L) group, and the 2 intervention groups pretreated with UA or DPI followed by stimulation with leptin for different times. The mRNA expression of Shh, Smo and Gli1/2 was detected in above cells after 12 hours’ treatment by RT-PCR. The protein expression of Rac1 and Gli2 were analyzed in the cells in 24 h of treatment by Western blotting. The proliferation of HSC-T6 cells was detected in 12, 24 and 48 h after treatment by MTT assay.       Results        RT-PCR analysis showed that Smo mRNA expression was increased when leptin stimulated HSC-T6 cells for 12 h (P<0.05). The expression of Shh and Gli2 mRNA was also increased compared with normal control group, but without significant difference. UA pretreatment significantly down-regulated the mRNA expression of Shh, Smo and Gli2 compared with control and leptin group (all P<0.05). Leptin, UA and DPI had no effect on Gli1 mRNA expression. The protein expression of Rac1 and Gli2 was increased when leptin stimulated HSC-T6 cells for 24 h (P<0.01). UA pretreatment significantly down-regulated Rac1 and Gli2 protein expression compared with leptin group (all P<0.01). MTT assay showed that leptin promoted the proliferation in HSC-T6 cells. After 12 hours’ pretreatment with UA, the cell growth was inhibited significantly than in leptin treated cells (P<0.01) in a time-dependent manner.       Conclusion        UA inhibits the expression of Shh, Smo, Gli2 mRNA and down-regulates the expression of Gli2 protein in HSC-T6 cells. One mechanism of UA inhibiting cell proliferation is probably via its inhibiting NOX subunit Rac1 and then NOX-Hh signaling pathway.

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更新日期/Last Update: 2014-03-03