复方中药861对大鼠肝星状细胞系一氧化氮合酶表达及其活性的影响

作者	单位
丁惠国	北京佑安医院消化科
王宝恩	北京友谊医院肝病研究中心
尚宏伟	首都医科大学综合实验室
王兴翠	首都医科大学综合实验室
唐素珍	北京友谊医院肝病研究中心
贾继东	北京友谊医院肝病研究中心
赵春惠	北京佑安医院消化科

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中文摘要:

目的 :研究复方中药 86 1对大鼠肝星状细胞系 (HSC T6 )一氧化氮合酶 (NOS)表达及其酶活性的影响 ,并探讨该药预防、治疗早期门脉高压的可行性。方法 :细胞密度为 1× 10 5/ml的HSC T6 接种于细胞培养皿中 ,95 %O2 ,5 %CO2 ,37℃ ,培养 2 4h进行以下分组实验 ,每组重复 6皿 ,继续培养 2 4h。 1组 :HSC T6 空白对照组 ;2组 :HSC T6 加复方中药 86 1(2mg/ml) ;3组 :HSC T6 加复方中药 86 1(4mg/ml) ;4组 :HSC T6 加复方中药 86 1(8mg/ml) ;5组 :HSC T6 加复方中药 86 1(4mg/ml)加L 硝基精氨酸甲酯 (L NAME ,4mg/ml)。采用化学比色法测定NOS活性 ,采用硝酸还原酶法测定培养液一氧化氮 (NO)水平。 4 %多聚甲醛固定细胞 2h ,采用免疫细胞化学法观察HSC T6 诱导型一氧化氮合酶 (iNOS)表达。结果 :复方中药 86 1能增加HSC T6 NOS活性 (P <0 0 1) ,上清液NO水平也平行增加 (P <0 0 1) ,L NAME不能抑制复方中药86 1刺激的HSC T6 合成、分泌NO增加 (P >0 0 5 )。免疫细胞化学研究发现 ,活化的HSC T6 细胞浆有iNOS表达 ,复方中药 86 1能增加HSC T6 iNOS表达。结论 :活化HSC T6 iNOS表达阳性 ,与自分泌NO有关 ;复方中药 86 1能明显增加HSC T6 iNOS表达及其酶活性 ,NO合成、分泌增加。复方中药 86 1可能通过增加肝

英文摘要:

Objective: To study the effect of herbal compound 861 (HB861) on expression and activity of nitric oxide synthase (NOS) in hepatic stellate cells (HSC), and to explore the feasibility of its application in preventing and treating the early portal hypertension. Methods: HSC of HSC T 6 cell line (1×10 5/ml) were cultured in dish with 95% O 2 plus 5% CO 2 under 37℃ for 24 hrs, then divided into 5 groups, 6 dishes in each group. Group A was the blank control group. To Group B E, HB861 2 mg/ml, HB861 4 mg/ml, HB861 8 mg/ml, HB861 4 mg/ml+N W Nitro L Arginine Methyl Ester (L NAME)4 mg/ml were added separately, and continuously cultured for 24 hrs. NOS activity was measured using colorimetry, NO level was determined by nitrate reductase technique. The cells were fixed by 4% paraformaldehyde for 2 hrs for test HSC T 6 iNOS expression by immuno cyto chemical method. Results: HB861 in 2mg/ml, 4mg/ml and 8mg/ml could increase HSC T 6 NOS activity from 1.7±0.1 to 2.5±0.3, 3.5±0.4 and 3.7±0.9 respectively (P<0 01), the NO levels in supernatant were increased in parallel from 56.1±4.8 to 90.7±4.6, 99.7±4.1 and 109.0±2.7 respectively (P<0 01). L NAME could not inhibit the effect of HB861 in increasing the synthesis and secretion of NO by activated HSC T 6. Immuno cyto chemical study showed that there was iNOS expression in cytoplasm, and which could be increased by HB861. Conclusion: The activated HSC T 6 showed positive iNOS expression, suggesting it could produce NO. HB861 could markedly increase HSC T 6 iNOS expression and NOS activity, enhance the NO synthesis and secretion, it also could inhibit the contractility of activated HSC by way of increase HSC to secrete NO, so as to lower the resistance in hepatic sinusoid, therefore would play important role in preventing and treating of early portal hypertension.

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