摘要
目的探究槲皮素对黄药子致肝损伤的保护作用及其机制。方法将48只昆明小鼠随机分为对照组、模型组、水飞蓟宾(100 mg/kg)组和槲皮素高、中、低剂量(80、50、20 mg/kg)组。对照组ig 0.5%羧甲基纤维素钠溶液,其余各组小鼠ig相应药物及黄药子醇提物(2 g/kg),1次/d,连续30 d。实验结束后称定小鼠体质量和肝脏质量,计算肝脏指数;测定血浆中天冬氨酸氨基转移酶(aspartate aminotransferase,AST)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、超氧化物歧化酶(superoxide dismutase,SOD)活性及丙二醛(malonaldehyde,MDA)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-10(interleukin-10,IL-10)、IL-1β水平;苏木素-伊红(hematoxylin-eosin staining,HE)染色观察肝脏组织病理学变化;TUNEL染色法检测肝脏细胞凋亡情况;采用Western blotting检测肝脏核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)、磷酸化核因子-κB p65(phosphorylated nuclear factor-κB p65,p-NF-κB p65)、血红素加氧酶-1(heme oxygenase-1,HO-1)、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)、多药耐药相关蛋白2(multidrug resistance-associated protein 2,MRP2)、尿苷二磷酸葡萄糖醛酸基转移酶1A1(uridine diphosphate-glucuronosyltransferase 1A1,Ugt1a1)、P-糖蛋白(P-glycoprotein,P-gp)表达;采用qRT-PCR检测肝脏MRP2、P-gp、Ugt1a1的mRNA表达。结果与模型组比较,槲皮素显著降低小鼠肝脏指数及肝功能(P<0.05、0.01),减轻肝脏组织病理变化,降低血浆中MDA水平(P<0.05、0.01),升高SOD活性(P<0.01),降低TNF-α、IL-1β水平(P<0.05、0.01),升高IL-10水平(P<0.01),抑制肝脏p-NF-κB p65、Bax蛋白表达(P<0.05、0.01),促进Nrf2、HO-1及药物转运体蛋白表达(P<0.05、0.01),并上调MRP2、P-gp、Ugt1a1的mRNA表达(P<0.01)。结论槲皮素对黄药子诱导的肝损伤有一定保护作用,其作用机制可能与减轻炎症、抑制细胞凋亡、调控Nrf2及药物转运体的表达相关。
Objective To investigate the protective effect and mechanism of quercetin against Dioscorea bulbifera(DB)-induced liver injury.Methods A total of 48 KM mice were randomly divided into control group,model group,silybin(100 mg/kg)group,and quercetin high-,medium-,low-dose(80,50,20 mg/kg)groups.Mice in control group was ig 0.5%CMC-Na solution.The remaining groups were administered with drug and ethanol extract(2 g/kg)of DB,once a day for 30 d.After the experiment,the body weight and liver weight were measured and liver index was calculated;Activities of aspartate aminotransferase(AST),alanine aminotransferase(ALT),superoxide dismutase(SOD),and levels of malonaldehyde(MDA),tumor necrosis factor-α(TNF-α),interleukin-10(IL-10),IL-1βin plasma were detected;Pathological changes in liver were observed by hematoxylin-eosin staining(HE);Apoptosis of liver cells was detected using TUNEL staining;Western blotting was used to determine protein expressions of nuclear factor erythroid 2-related factor 2(Nrf2),phosphorylated nuclear factor-κB p65(p-NF-κB p65),heme oxygenase-1(HO-1),B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),multidrug resistance-associated protein 2(MRP2),uridine diphosphate-glucuronosyltransferase1A1(Ugt1a1)and P-glycoprotein(P-gp)in liver;qRT-PCR was used to detect mRNA expressions of MRP2,P-gp and Ugt1a1 in liver.Results Compared with model group,quercetin significantly reduced liver index and liver function(P<0.05,0.01),reduced pathological change in liver tissue,reduced MDA level in plasma(P<0.05,0.01),increased SOD activity(P<0.01),reduced levels of TNF-α,IL-1β(P<0.05,0.01),increased IL-10 level(P<0.01),inhibited p-NF-κB p65 and Bax protein expressions in liver(P<0.05,0.01),promoted Nrf2,HO-1 and drug transporter protein expressions(P<0.05,0.01),and up-regulated the mRNA expressions of MRP2,P-gp and Ugt1a1(P<0.01).Conclusion Quercetin has a certain protective effect on DB-induced liver injury,and its mechanism may be related to reducing inflammation,inhibiting cell apoptosis,regulating the expression of Nrf2 and drug transporters.
作者
王欣
张雨涵
史磊磊
上官惠子
宋佳莲
安宇宇
谢允东
刘继平
WANG Xin;ZHANG Yuhan;SHI Leilei;SHANGGUAN Huizi;SONG Jialian;AN Yuyu;XIE Yundong;LIU Jiping(Shaanxi University of Chinese Medicine,Xianyang 712046,China;Shaanxi Key Laboratory for Safety Monitoring of Food and Drug,Xianyang 712046,China;Key Laboratory of Pharmacodynamic Mechanism and Material Basis of Chinese Medicine of Shaanxi Administration of Traditional Chinese Medicine,Xianyang 712046,China)
出处
《中草药》
CAS
CSCD
北大核心
2024年第10期3345-3353,共9页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金资助项目(82204790)。
关键词
槲皮素
黄药子
肝损伤
炎症
凋亡
核因子E2相关因子2
药物转运体
quercetin
Dioscorea bulbifera L.
liver injury
inflammation
apoptosis
nuclear factor erythroid 2-related factor 2
drug transporter
