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目的:评价新型肝靶向一氧化氮释放药物NO-040527抗小鼠肝损伤的药效学及细胞毒性,为进一步的临床前研究开发奠定基础.方法:通过腹腔注射四氯化碳、D-氨基半乳糖或对乙酰氨基酚,制备肝损伤动物模型;分别于造模前1h和造模后12h灌胃给予不同剂量的待测药物,于第2次给药后24h,眼眶取血,留取血清标本,测定血清中的ALT,AST水平.取对数生长期的HepG2细胞,接种于96孔培养板,长成单层,加入不同浓度的待测药物.继续培养24h,用MTT法,测定细胞的存活率.结果:肝靶向药物NO-040527能够显著降低CCl4所致小鼠肝损伤的转氨酶升高(P<0.01),降酶作用具有良好的剂量依赖性;在相同剂量(55mg/kg)下,NO-040527的降酶作用强于阳性对照药NCX-1000(P<0.05).同样,肝靶向药物NO-040527能够显著降低对乙酰氨基酚所引起的小鼠转氨酶升高(P<0.01),各剂量组之间降酶作用无显著性差异,在相同剂量(55mg/kg)下,NO-040527的降酶作用与NCX-1000相当.NO-040527对D-氨基半乳糖所致小鼠肝损伤没有保护作用,中低剂量组的小鼠血清ALT、AST值较模型组相比没有显著差异,高剂量组转氨酶甚至略高于模型组.NO-040527在500μmol/L浓度显示出细胞毒性,细胞存活率为45.96%±29.46%(P=0.058);在1000μmol/L浓度具有显著的细胞毒性(P<0.005);在50μmol/L和100μmol/L浓度时,NO-040527显示出促细胞生长的作用,细胞存活率分别为137.67%±8.47%和152.65%±10.084%,显著高于溶剂组(P<0.05).结论:NO-040527能够显著降低CCl4和对乙酰氨基酚所引起的小鼠转氨酶升高,但NO-040527和NCX-1000对D-氨基半乳糖所致小鼠肝损伤没有保护作用,高剂量甚至加重转氨酶升高.NO-040527在高浓度显示出细胞毒性,低浓度显示出促细胞生长的作用.
OBJECTIVE: To evaluate the pharmacodynamics and cytotoxicity of NO-040527, a novel liver-targeted nitric oxide releasing drug, against liver injury in mice and lay a foundation for further preclinical research and development.Methods: The intraperitoneal injection of carbon tetrachloride, D- Galactosamine or acetaminophen to prepare animal model of hepatic injury. Different doses of test drug were given intragastrically at 1h before model making and at 12h after model making, respectively. Blood was collected from the orbital orifice after 24h Serum samples were taken and serum levels of ALT and AST were measured.HepG2 cells in logarithmic growth phase were inoculated into 96-well culture plates and grown into monolayer and different concentrations of drugs to be tested were added.Using MTT method, The survival rate of cells was measured.Results: The liver-targeting drug NO-040527 could significantly reduce the transaminase of liver injury induced by CCl4 in mice (P <0.01) / kg), NO-040527 showed a stronger antihypertensive activity than NCX-1000 (P <0.05). Similarly, the liver-targeting drug NO-040527 significantly reduced acetaminophen-induced elevation of transaminases (P <0.01). There was no significant difference in the activity of down-regulating enzymes between different dosage groups. Under the same dose (55mg / kg) NO-040527 had similar hypnagase activity as NCX-1000. NO-040527 had no protective effect on D-galactosamine-induced liver injury in mice, and serum ALT and AST in low and middle dose groups were lower than those in model group (P = 0.058). At the concentration of 1000 μmol / L, NO-040527 showed cytotoxicity at a concentration of 500 μmol / L, with a cell viability of 45.96% ± 29.46% (P <0.005). NO-040527 showed the effect of promoting cell growth at the concentration of 50μmol / L and 100μmol / L, the cell survival rate was 137.67% ± 8.47% and 152.65% ± 10.084% respectively, which was significantly higher than Solvent group (P <0.05) .Conclusion: NO-040527 can significantly reduce the elevated transaminases induced by CCl4 and acetaminophen in mice, but NO-040527 and NCX- No protective effect of injury, high doses and even increased elevated transaminases.NO-040527 showed cytotoxicity at high concentrations, low concentrations showed the role of pro-cell growth.