Xiao Yong-Hong, Liu Dian-Wu, Li Qing
Department of Epidemiology, Hebei Medical University, Shijiazhuang 050017, Hebei Province, China.
World J Gastroenterol. 2005 Mar 14;11(10):1515-20. doi: 10.3748/wjg.v11.i10.1515.
To investigate the effects of anti-fibrosis I herbal compound on intracellular Ca(2+) in activated hepatic stellate cell (HSC) and to try to survey its molecular mechanism in treatment and prevention of hepatic fibrosis and portal hypertension.
The activated HSC line was plated on small glass cover slips in 24 wells culture dishes at a density of 5X10(6) /mL, and incubated in RPMI-1640 media for 24 h. After the cells were loaded with Fluo-3/AM, intracellular Ca(2+) was measured with laser scanning confocal microscopy (LSCM). The dynamic changes of intracellular Ca(2+), stimulated by carbon tetrachloride, TGF-beta(1) antibody and the drug serum of anti-fibrosis I herbal compound and under orthogonal design were determined by LSCM. The effect of anti-fibrosis I herbal compound on intracellular Ca(2+) was observed before and after the addition of TGF-(1) antibody.
The intracellular Ca(2+) were significantly different in different dosage of carbon tetrachloride anti-fibrosis I formula drug serum, TGF-beta(1) antibody and different turn of these substance, but their interval time between CCl(4) and TGF-beta(1) antibody, CCl(4) and anti-fibrosis I drug serum had no influence on intracellular Ca(2+). The result showed intracellular Ca(2+) wasn't significantly different between rat serum without anti-fibrosis I and untreated group. After carbon tetrachloride stimulation, intracellular Ca(2+) of activated HSC increased significantly when the dosage of CCl(4) from 5 to 15 mmol/L, however, decreased significantly after stimulation by 5-20 microg/mL TGF-beta(1) antibody or 5-20 mL/L drug serum. Moreover, before and after the addition of TGF-beta(1) antibody, intracellular Ca(2+) was significantly different. These results suggested that the molecular mechanism was independent of blocking TGF-beta(1) effects.
Anti-fibrosis I herbal compound may treat hepatic fibrosis and decrease portal hypertension by inhibiting activated HSC contractility through decrease of intracellular Ca(2+).
研究抗纤维化I号中药复方对活化肝星状细胞（HSC）内钙离子的影响，并探讨其在治疗和预防肝纤维化及门静脉高压中的分子机制。
将活化的HSC细胞系以5×10⁶/mL的密度接种于24孔培养板中的小玻璃盖玻片上，在RPMI-1640培养基中培养24小时。用Fluo-3/AM负载细胞后，用激光扫描共聚焦显微镜（LSCM）测量细胞内钙离子浓度。通过LSCM测定在四氯化碳、转化生长因子-β1（TGF-β1）抗体和抗纤维化I号中药复方含药血清刺激下，以及在正交设计条件下细胞内钙离子的动态变化。在加入TGF-β1抗体前后观察抗纤维化I号中药复方对细胞内钙离子的影响。
不同剂量的四氯化碳、抗纤维化I号复方含药血清、TGF-β1抗体以及这些物质的不同加入顺序，细胞内钙离子浓度均有显著差异，但四氯化碳与TGF-β1抗体、四氯化碳与抗纤维化I号含药血清之间的间隔时间对细胞内钙离子浓度无影响。结果显示，不含抗纤维化I号的大鼠血清组与未处理组细胞内钙离子浓度无显著差异。四氯化碳刺激后，当四氯化碳剂量从5 mmol/L增加到15 mmol/L时，活化HSC的细胞内钙离子浓度显著升高，然而，在5 - 20 μg/mL TGF-β1抗体或5 - 20 mL/L含药血清刺激后显著降低。此外，加入TGF-β1抗体前后，细胞内钙离子浓度有显著差异。这些结果表明其分子机制独立于阻断TGF-β1的作用。
抗纤维化I号中药复方可能通过降低细胞内钙离子浓度抑制活化HSC的收缩性，从而治疗肝纤维化并降低门静脉高压。
