黄芩苷抑制脂多糖促巨噬细胞氧化应激损伤作用的研究  

Role of baicalin and mechanisms through which baicalin attenuates oxidative stress injury induced by lipopolysaccharide on macrophages

作　　者：黄伟琨[1] 徐秋艳 周婷[1] Huang Weikun;Xu Qiuyan;Zhou Ting(Dept.of Stomatology,Guizhou Provincial People’s Hospital,Guiyang 550002,China)

机构地区：[1]贵州省人民医院口腔科,贵阳550002

出　　处：《国际口腔医学杂志》2022年第5期521-528,共8页International Journal of Stomatology

基　　金：国家自然科学基金(81760198);贵州省中医药、民族医药科学技术研究专项课题(QZYY-2019-011);贵州省人民医院青年基金(GZSYQN[2018]08,GZSYQN[2018]10);贵州省卫生健康委员会科学技术基金(gzwjkj2018-1-054);贵州省人民医院国家自然科学基金培育基金(QKHPTRC[2018]5764-06)。

摘　　要：目的研究黄芩苷在牙龈卟啉单胞菌(P.gingivalis)的致病因子脂多糖(LPS)促巨噬细胞氧化应激反应中的作用及分子机制。方法LPS刺激巨噬细胞的同时,采用不同浓度(5、10μmol·L^(-1))的黄芩苷进行干预,分别采用细胞活性检测试剂盒(CCK8)、乳酸脱氢酶(LDH)试剂盒、2,7-二氢二氯荧光黄(DCFA-DA)探针、丙二醛(MDA)、超氧化物歧化酶(SOD)试剂盒、流式细胞仪,检测P.gingivalis-LPS对细胞所造成的损伤、细胞内活性氧(ROS)含量、MDA和SOD活性、细胞凋亡;随后,采用蛋白质印迹法(WB)检测核因子E2相关因子2(Nrf2)的总蛋白、胞质蛋白及胞核蛋白的表达情况。结果与对照组相比,P.gingivalis-LPS可以导致细胞活力下降(P<0.0001)、LDH含量上升(P<0.0001)、ROS和MDA含量上调(P<0.0001)、SOD活性下降(P<0.0001)以及细胞凋亡率上升(P<0.0001)。黄芩苷的干预能够减轻P.gingivalis-LPS对细胞所造成的上述损伤(P<0.0001);10μmol·L^(-1)黄芩苷的干预能够显著上调胞核Nrf2的表达(P<0.01)。结论黄芩苷通过促进巨噬细胞Nrf2的核转位,减轻了P.gingivalis-LPS所诱发的氧化应激损伤。Objective We aimed to explore the role and mechanisms of baicalin in oxidative injury promoted by Porphyromonas gingivalis(P.gingivalis)lipopolysaccharides(LPSs)on macrophages.Methods Macrophages derived from human monocytic-leukemia cells(THP-1)were stimulated with LPSs with or without baicalin(5 and 10μmol·L^(-1)),and then cell injury,reactive oxygen species(ROS)level,malondialdehyde(MDA),superoxide dismutase(SOD)activities,and apoptosis were measured through cell counting kit-8(CCK8),lactate dehydrogenase(LDH),2,7-dichlorodihydrofluorescein diacetate,MDA,and SOD assays;enzyme-linked immunosorbent assay;and flow cytometry.The expression level of nuclear factor erythroid 2-related factor 2(Nrf2)was determined by Western Blot(WB).Results LPS inhibited cell proliferation(P<0.0001),induced LDH leakage(P<0.0001),increased ROS level and MDA activity(P<0.0001),decreased SOD activity(P<0.0001),and increased apoptotic rate(P<0.0001).Baicalin co-treatment attenuated damage induced by LPSs(P<0.0001).Meanwhile,the cytoplasmic protein level of Nrf2 was up-regulated by 10μmol·L^(-1) baicalin co-treatment(P<0.01).Conclusion Baicalin attenuated LPS-promoted oxidative injury on macrophages by up-regulating Nrf2.

关 键 词：黄芩苷 牙周炎 氧化应激 巨噬细胞 

分 类 号：Q26[生物学—细胞生物学]