内皮素-1在脂多糖致大鼠门脉高压症中作用的体外研究

Role of endothelin in portal hypertension induced by endotoxin

目的:研究内皮素-1(ET-1)在脂多糖致门脉高压中的作用及其机制。方法:分离、纯化大鼠星状细胞,培养在胶原凝胶上,应用ET-1-ASON和LPS序贯共培养(实验组),以正义、错义ASON作为对照,检测凝胶收缩情况;应用放射免疫法检查培养上清液的ET-1浓度;免疫印迹法检测各组细胞α-actin表达情况;RT-PCR检测各组细胞ET-1表达情况。结果:实验组的孔胶原直径缩小至93.3%±3.8%,胶原直径较对照组大(P<0.05);实验组HSC培养基上清液ET-1为(49.8±7.4)ng/L,显著低于对照组(P<0.01);实验组HSC表达β-ac-tin较对照组弱;实验组HSC表达ET-1 mRNA显著低于对照组(P<0.01)。结论:ET-1在脂多糖致门脉高压中通过抑制星状细胞活化而起重要作用,ET-1-ASON在控制脂多糖致门脉高压中具有应用前景。

AIM： To study the role of endothelin - 1 （ ET - 1 ） in portal hypertension （PHT） induced by endotoxin. METHODS： Collagenase in situ perfusion was adopted to separate hepatic stellate cells （HSCs）. HSCs was cultured on concretized collagen. ET - 1 anti - sense oligonucleotide was added into the culture medium and then LPS was also added up to the concentration of 1 000 ug/L. The diameters of the concretized collagen were measured. Sense and mis - sense oligonucleotide were applied as control. ET - 1 in the culture medium was detected by radioimmunoassay and ET - 1 mRNA in HSCs was detected by RT - PCR. β- actin of HSCs was detected by Western blotting. RESULTS： The diameter of concretized collagen on which HSCs pretreated with ET - 1 anti - sense oligonucleotide was 93. 3% ± 3. 8% the size of the primary. The diameter of concretized collagen of the control groups were 70. 1% ±4. 8% and 70. 5% ±3.9% （P 〈0. 05）. ET - 1 was （49. 8 ＋ 7.4） ng/L in the culture medium of HSCs pretreated with ET - 1 anti - sense oligonucleotide and （ 329. 8 ± 34. 9）, （339. 1 ± 43. 7） ng/L in the control medium （ P 〈 0. 05 ）. β - actin and ETI mRNA presented in HSCs pretreated with ET - 1 anti - sense oligonucleotide was much less than that in the controls. CONCLUSION： ET - 1 anti - sense oligonueleotide inactivated HSCs by counteracting the expression of ET - 1, which may be helpful to control PHT induced by LPS.