摘要
目的:研究八宝丹(BBD)对对乙酰氨基酚(APAP)诱导的急性肝损伤小鼠NOD样受体热蛋白结构域相关蛋白3/胱天蛋白酶-1(NLRP3/Caspase-1)通路蛋白的影响。方法:将C57BL/6小鼠随机分组,BBD(75、150、300 mg·kg^(-1))灌胃给药3 d,2次/d,于末次给药后2 h,腹腔注射APAP(400 mg·kg^(-1)),14 h后摘眼球取血,随后脱颈椎处死取材。苏木素-伊红(HE)染色法观察肝组织细胞形态学变化;生化法检测各组小鼠血清中丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、超氧化物歧化酶(SOD)、丙二醛(MDA)和髓过氧化物酶(MPO)的活性;实时荧光定量聚合酶链式反应(Real-time PCR)检测小鼠肝脏中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)及白细胞介素-6(IL-6) mRNA表达水平;蛋白免疫印迹法(Western blot)检测小鼠肝脏中环氧化酶-2(COX-2)、诱导型一氧化氮合酶(iNOS)、NLRP3、Caspase-1、白细胞介素-18(IL-18)的蛋白表达水平。结果:与正常组比较,模型组小鼠肝脏组织小叶结构部分破坏、肝窦扩张,血清中ALT和AST水平,肝组织中NLRP3、Caspase-1、iNOS、IL-18、COX-2蛋白表达水平和IL-1β、IL-6、TNF-α mRNA水平均明显升高(P<0.05,P<0.01)。与模型组比较,各给药组小鼠肝细胞破裂现象有所好转,肝血窦挤压情况有所缓解,血清中ALT和AST水平,肝组织中NLRP3、Caspase-1、iNOS、IL-18、COX-2蛋白表达水平和IL-1β、IL-6、TNF-α mRNA水平均明显降低(P<0.05,P<0.01),并呈剂量依赖性。结论:BBD可减轻APAP诱导的小鼠急性肝损伤,其机制可能与抗氧化应激,抑制NLRP3/Caspase-1通路,进而降低IL-1β、IL-18、TNF-α、IL-6表达水平有关。
Objective:To explore the effect of Babaodan(BBD) on the NOD-like receptor pyrin domain containing 3/cysteine aspartate-specific protease-3(NLRP3/Caspase-1) pathway proteins in mice with acetaminophen(APAP)-induced acute liver injury. Method:C57BL/6 mice were randomly grouped, and BBD(75, 150, 300 mg·kg^(-1), ig) was administered twice a day for three days. After 2 hours of the last administration, the mice were treated with APAP(400 mg·kg^(-1), ip), and the eyeballs were removed to collect blood after 14 hours. Then they were sacrificed by cervical dislocation for sample collection. Hematoxylin-eosin(HE) staining was used to observe the morphological changes of liver tissue cells, and biochemical methods were used to detect the activities of alanine aminotransferase(ALT), aspartate aminotransferase(AST), superoxide dismutase(SOD), malondialdehyde(MDA) and myeloperoxidase(MPO) in serum of mice in each group. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was performed to determine the mRNA expression of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) and IL-6, and Western blot was performed to determine the protein expression of cyclooxygenase-2(COX-2), inducible nitric oxide synthase(iNOS), NLRP3, Caspase-1 and IL-18 in the liver of mice. Result:Compared with the conditions in normal group, the hepatic lobule structure of mice in the model group was partially destroyed, and the hepatic sinusoids were dilated. And the expression levels of ALT and AST in serum, the protein levels of NLRP3, Caspase-1, iNOS, IL-18 and COX-2 and the mRNA levels of IL-1β, IL-6 and TNF-α were increased(P<0.05, P<0.01). Compared with the model group, the administration groups had improvement in liver cell rupture and hepatic sinusoidal compression, and a dose-dependent decrease in the levels of ALT and AST in serum as well as the protein levels of NLRP3, Caspase-1, iNOS, IL-18 and COX-2 and the the mRNA levels of IL-1β, IL-6 and TNF-α in liver tissue(P<0.05, P<0.01). Conclusion:BBD can reduce APAP-induced acute liver injury in mice. The mechanism may be related to anti-oxidative stress, inhibition of NLRP3/Caspase-1 pathway, and decreased expression levels of IL-1β, IL-18, TNF-α and IL-6.
作者
赵若玮
张庆
朱铭星
刘跃洋
程再兴
黄鸣清
郑燕芳
林彦翔
ZHAO Ruowei;ZHANG Qing;ZHU Mingxing;LIU Yueyang;CHENG Zaixing;HUANG Mingqing;ZHENG Yanfang;LIN Yanxiang(College of Pharmacy,Fujian University of Traditional Chinese Medicine(TCM),Fuzhou 350122,China;College of Traditional Chinese Medicine,Fujian University of TCM,Fuzhou 350122,China;College of Acupuncture and Moxibustion,Fujian University of TCM,Fuzhou 350122,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第5期122-128,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81973437)。
