摘要
为了探究大黄黄芩药对对脂多糖(lipopolysaccharides,LPS)诱导的内毒素血证模型大鼠肝脏炎症的调节机制,选取取雄性SD大鼠50只,随机分为空白组、模型组、地塞米松组、药对高剂量组、药对低剂量组,每组10只。各组大鼠给予相应药物进行预防性给药,连续给药7 d。末次给药结束0.5 h后尾静脉注射LPS(5 mg·kg^(-1))造模,后每0.5 h测定一次动物肛温并记录,于造模后4 h处死动物,测定脾脏胸腺系数;采用ELISA法检测肝组织中细胞因子白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的含量;采用比色法测定肝组织中一氧化氮(NO)含量;采用Western blot法检测肝组织中Toll样受体蛋白4(Toll样受体-4),p38MAPK,磷酸化p38MAPK(p-p38MAPK)及一氧化氮合成酶(i NOS)蛋白相对表达量。结果显示,与空白组大鼠相比,模型组大鼠肝组织中TLR4蛋白表达、iNOS蛋白表达、p38磷酸化表达升高,IL-1β,NO,TNF-α含量显著升高(P<0.05或P<0.01)。与模型组比较,药对高剂量能显著降低大鼠肝组织中IL-1β,NO,TNF-α的表达(P<0.05或P<0.01),下调i NOS蛋白表达与p38磷酸化表达(P<0.05),但对TLR4蛋白并未出现显著的回调作用;药对低剂量能显著降低模型大鼠肝组织中IL-1β,NO的表达(P<0.01),显著下调i NOS蛋白表达(P<0.01),对p38磷酸化表达具有一定的下调趋势但不具有统计学意义,而对TLR4蛋白表达并没有表现出一定降低作用。大黄黄芩配伍使用可能是通过减少p38蛋白的磷酸化表达从而阻断p38 MAPK信号通路,来减少i NOS蛋白表达和炎性因子IL-1β,NO,TNF-α的释放,从而达到减缓炎症反应保护机体组织的作用。
To explore the mechanism of Rhei Radix et Rhizoma combined with Scutellariae Radix in regulatory lipopolysaccharide （LPS） -induced liver inflammation in rats with endotoxin blood, 50 male SD rats were selected and randomly divided into blank group, model group, dexamethasone group, herbal pair high dose group, herbal pair low dose group, with 10 in each group. Rats in each were given preventive drugs for 7 consecutive days. At 0. 5 h after the final administration, the model was built through the tail vein injection with LPS （5 mg ~ kg- 1 ）. Then, animal anal temperatures were determined and recorded once every 0. 5 h. The rats were killed at 4 h after the modeling to determine spleen thymus coefficient. ELISA method was used to detect cytokines interleukin-1/3 （IL-1/~） and tumor necrosis factor-alpha （TNF-α） in liver tissues. The colorimetrie method was applied in determination of oxidation nitrogen （NO） content in liver tissues. Western Blot method was adopted to detect Toll-like receptor protein 4, p38MAPK p38MAPK, phosphorylated p38MAPK （p-p38MAPK） and nitric oxide synthase （iNOS） protein expressions. The results showed that compared with the blank group, in the model group, TLR4 protein expression, iNOS protein expression and p38 phosphorylation expression, IL-lfl, NO and TNF-a content increased significantly in liver tissues （P 〈 0. 05 or P 〈 0. 01 ）. And compared with the model group, the herbal pair high dose group showed significantly reduction in IL-lfl, NO and TNF-a expressions in rat liver tissues （P 〈 0. 05 or P 〈 0. 01 ） , down- regulation in iNOS protein expression and p38 phosphorylation expression in rat liver tissues （ P 〈 0. 05 ）, but without significant up- regulation in TLR4 protein. Low-dose herbal pair can significantly reduce IL-lfl and NO expression in model rat liver tissues （ P 〈 0. 01 ）, significantly down-regulate iNOS protein expression （ P 〈 0. O1 ） , with a slight down-regulation in phosphorylation of p38 but no statistical significance, and no reduction in TLR4 expression. In conclusion, the compatibility of Rhei Radix et Rhizoma combined with Scutellariae Radix may reduce the expression of iNOS protein and the release of inflammatory cytokines IL-1β, NO and TNF-a by de- creasing p38 protein phosphorylation expression and blocking p38MAPK signaling pathways, so as to alleviate the inflammation reaction and protect the liver.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2016年第18期3425-3431,共7页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81274111)
成都市科技项目(12DXYB239JH-002)
