益肝康等活血化瘀中药抑制IL-1β刺激的HSC增殖及TIMP-1的表达

Yigankang inhibits proliferation of hepatic stellate cells and expression of TIMP-1 mRNA induced by interleukin-1β

目的:探讨活血化瘀中药益肝康、丹参小复方、丹参对IL-1β刺激的活化的大鼠肝星状细胞(HSCs)增殖及基质金属蛋白酶抑制因子 mRNA(TIMP mRNA)表达的影响．方法:体外培养活化的大鼠HSC,随机分为8 组:对照组(A组)、IL-1β 10 μg/L(B组)、IL- 1β 10μg/L+益肝康2 g/L干预组(C组)、IL- 1β 10 μg/L+丹参小复方2g/L干预组(D组)、 IL-1β 10μg/L+丹参2g/L干预组(E组)、丹参 2 g／L(F组)、丹参小复方2 g/L(G组)、益肝康 2 g／L(H组)．加药后24 h．应用活细胞计数试剂盒-CCK-8检测各组HSC增殖,采用半定量 RT-PCR方法检测各组HSC TIMP-1 mRNA的表达．结果:A组HSC增殖和TIMP-1 mRNA表达强于F、G、H组(1．291±0．09 vs 1．055±0．105, 1±0．07,0．883±0．06,P<0．01:0．591±0．064 vs 0．493±0．088．0．458±0．076．0．356±0．046． P<0．05或P<0．01);H组HSC增殖和TIMP-1 mRNA表达低于F组和G组(P<0．05);B组HSC 增殖和TIMP-1 mRNA表达均明显强于A组 (1．575±0．017 vs 1.291±0,09,P<0．01;1．369± 0．097 vs 0．591±0．064,P<0．01)和C、D、E组 (1．575±0．017 vs 0．906±0．09,1．015±0．081, 1．097±0．038,P<0．01;1．369±0．097 vs 0．694 ±0．078,0．854±0．05,0．898±0．12,P<0．01);C 组HSC增殖和TIMP-1 mRNA表达低于D组和 E组(P<0．05)．结论:益肝康等活血化瘀中药能抑制IL-1β刺激的HSCs增殖及TIMP-1 mRNA表达,发挥其抗肝纤维化功效．益肝康抗肝纤维化作用强于丹参小复方和丹参单药．

AIM： To investigate the effects of the bloodactivating and stasis-eliminating traditional Chinese medicines （TCM）, including Yigankang, small compound of Radix Salviae Miltiorrizae （scRSM）, and Radix Salviae Miltiorrizae （RSM）, on the proliferation of rat hepatic stellate cells （HSCs） and expression of tissue inhibitor of matrix metalloproteinase-1 （TIMP-1） mRNA induced by interleukin-1β （IL-1β）. METHODS： The activated rat HSCs were cul- tured in vitro and then randomly divided into 8 groups, named A, B, C, D, E, F, G, and H. The rats in group A served as the controls, and those in the rest groups were treated IL-1β （10 μg/L）, IL-1β （10μg/L） plus Yigankang （2 g/L）, IL-1β （10 μg/L） plus scRSM （2 g/L）, IL-1β （10 μg/L） plus RSM （2 g/L）, RSM （2 g/L）, scRSM （2 g/L）, and Yigankang （2 g/L）, respectively. The proliferation of HSCs was detected by cell counting kit-8 （CCK-8） and the expression of the TIMP-1 mRNA was detected by semi-quantitative reverse transcription chain reaction （RT-PCR）. RESULTS： The proliferation of HSCs and expression of TIMP-1 mRNA in group A were significantly higher than those in group F, G, H （1.291 ± 0.09 vs 1.055 ± 0.105, 1 ± 0.07, 0.883 ± 0.06, P 〈 0.01; 0.591 ± 0.064 vs 0.493 ± 0.088, 0.458 ± 0.076, 0.356 ± 0.046, P 〈 0.05 or P 〈 0.01）, and they were markedly lower in group H than those in group F and G （P 〈 0.05）. The proliferation of HSCs and expression of TIMP-1 mRNA in group B were significantly increased in comparison with those in group A, C, D, and E （1.575 ± 0.017 vs 1.291 ± 0.09, 0.906 ± 0.09, 1.015 ± 0.081, 1.097 ± 0.038, P 〈 0.01; 1.369 ± 0.097 vs 0.591 ± 0.064, 0.694 ± 0.078, 0.854 ± 0.05, 0.898 ± 0.12, P 〈 0.01）, but they were notably increased in group C than those in group D and E （P 〈 0.05）. CONCLUSION： Yigankang as well as scRSM and RSM can inhibit the IL-1β-induced proliferation of HSCs and expression of TIMP-1 mRNA, by which it plays a protective role against liver inflammation and fibrosis. Yigankang is more effective than scRSM and RSM.