氧化苯砷体外对大鼠原代肝星状细胞活化的阻抑作用研究*

doi:10.3969/j.issn.1672-5069.2016.04.008

实用肝脏病杂志 ›› 2016, Vol. 19 ›› Issue (4): 413-417.doi: 10.3969/j.issn.1672-5069.2016.04.008

氧化苯砷体外对大鼠原代肝星状细胞活化的阻抑作用研究^*

仝艳艳, 李光明, 邓怡林, 石翠翠, 黄福德, 范建高

200092 上海市交通大学医学院附属新华医院消化内科（仝艳艳,李光明,石翠翠,邓怡林,范建高）;
上海科学院高等研究院神经生物学研究组（黄福德）

收稿日期:2015-12-09 出版日期:2016-07-30 发布日期:2016-08-31
通讯作者: 李光明,E-mail: ligm68@126.com
作者简介:仝艳艳,女,24,硕士研究生。主要从事肝损伤与肝纤维化的防治研究。E-mail:tyanyan1991@163.com
基金资助:
国家自然科学基金资助项目（81070344/81400631）; 中国肝炎基金会王宝恩肝纤维化研究基金资助项目（200090007）

Inhibition of primary cultured hepatic stellate cell activation by phenylarsine oxide in vitro

Tong Yanyan, Li Guangming, Deng Yilin, et al

Department of Gastroenterology,Xinhua Hospital Affiliated to JiaoTong University School of Medicine,Shanghai 200092

Received:2015-12-09 Online:2016-07-30 Published:2016-08-31

摘要/Abstract

摘要： 目的探讨氧化苯砷(PAO)对肝星状细胞（HSC）活化的影响。方法采用密度梯度离心法提取SD大鼠原代HSC,在倒置显微镜下观察HSC形态的改变;以25、50、100、150和200 nmol/L浓度PAO处理活化的HSC-T6细胞,以四甲基偶氮唑盐（MTT）法评估PAO的细胞毒性;分别以25、50、100 nmol/L浓度的PAO处理离体培养4 d的HSC 72 h,并设对照组,采用Western blot和Real-time PCR法检测各组细胞α-SMA和I型胶原mRNA和蛋白表达。结果原代HSC离体培养过程中α-SMA表达量逐渐升高,与培养1 d时（0.762±0.062）比,培养4 d时其α-SMA蛋白表达【(1.51±0.045),P<0.05】显著升高;PAO在25~100 nmo/L浓度范围内对活化的HSC无明显的细胞毒性,但能浓度依赖性抑制活化的HSC α-SMA 和I型胶原mRNA水平和蛋白表达。结论离体培养4 d时,HSC呈初始活化状态。PAO在25~100 nmol/L范围可浓度依赖性地阻抑离体培养的HSC的自发激活,提示PAO有潜力成为一类新型的抗肝纤维化药物。

关键词: 肝纤维化, 肝星状细胞, 氧化苯砷, 体外

Abstract: Objective To investigate the inhibition of primary cultured hepatic stellate cell （HSC） activation by phenylarsine oxide （PAO） in vitro. Methods Primary cultured HSCs were isolated from rat 1iver by Nycodenz density-gradient centrifugation. The morphological features of the cells were observed under inverted microscope. The cytotoxicity of PAO on HSCs-T6 was determined by MTT after treated with 5 different PAO concentrations（25,50,100,150 and 200 nmol/L） for 24 h. We chose proper concentration of PAO （<100 nmol/L） in this experiment,and then we divided primary cultured HSCs at day 4 into four groups receiving PAO at concentration of 0,25,50 and 100 nmol/L respectively. The expressions of α-SMA and type I collagen were measured by Western blot and Real-time PCR respectively. Results Expression of α-SMA was up-regulated as HSCs activation at primary culture （the expression of α-SMA increased significantly at day 4 （1.51±0.045） as compared with at day 1[（0.762±0.062）,P<0.05】; PAO at 25 to 100 nmol/L concentration range had no obvious cytotoxicity to activated HSCs,and down-regulated and inhibited α-SMA mRNA and collagen type I mRNA levels and their protein expression of HSCs in a concentration-dependent manner. Conclusion The HSCs is initially activated at day four in vitro culture,and PAO at 25 to 100 nmol/L range can inhibit the spontaneous activation of HSCs in vitro.

Key words: Hepatic fibrosis, Hepatic stellate cells, Phenylarsine oxide, In vitro

仝艳艳, 李光明, 邓怡林, 石翠翠, 黄福德, 范建高. 氧化苯砷体外对大鼠原代肝星状细胞活化的阻抑作用研究^*[J]. 实用肝脏病杂志, 2016, 19(4): 413-417.

Tong Yanyan, Li Guangming, Deng Yilin, et al. Inhibition of primary cultured hepatic stellate cell activation by phenylarsine oxide in vitro[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2016, 19(4): 413-417.

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氧化苯砷体外对大鼠原代肝星状细胞活化的阻抑作用研究^*

Inhibition of primary cultured hepatic stellate cell activation by phenylarsine oxide in vitro

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