摘要
目的研究冰片对脂多糖(LPS)引发脑微血管内皮细胞(BMECs)损伤的保护作用及潜在机制。方法原代培养和鉴定大鼠BMECs,并用LPS诱发炎性损伤。利用CCK-8法检测细胞的存活率,并以此优化冰片的给药剂量。继而通过ELISA检测TNF-α、IL-6和IL-8的生成,DCFH-DA探针检测ROS含量,Hoechst 33342染色检测细胞凋亡率,Western blot检测TLR4、p-p65、p65、p-IκBα和IκBα的表达。结果冰片剂量优化结果表明其在10 mg·L^(-1)和20 mg·L^(-1)具有良好的剂量-效应依赖性。以它们为低、高剂量,发现冰片可显著减少TNF-α、IL-6和IL-8的分泌和ROS的生成,降低凋亡细胞百分率,减少TLR4、p-p65和IκBα的表达,并增加p65和p-IκBα的表达。结论冰片可通过下调TLR4-NF-κB通路缓解BMECs的炎症反应,进而减少大脑损伤。
The study was performed to investigate the protection of borneol against LPS-induced brain microvascular endothelial cells(BMECs)injury and the underlying mechanism.Rat BMECs were primarily cultured and identified,and then induced by LPS to construct inflammatory injury model.CCK-8 kit was used to detect the cell survival rate and optimize the dosage of borneol.The secretion of TNF-α,IL-6 and IL-8 were measured by ELISA,the content of ROS was detected by DCFH-DA probe,the apoptosis ratio was assessed by Hoechst 33342 staining,and the expression of TLR4,p-p65,p65,p-IκBαand IκBαwere tested by Western blotting.Doses optimization result indicated that 10 mg·L^(-1) and 20 mg·L^(-1) of borneol displayed a dose-effect relationship.At these two doses,borneol significantly decreased the secretion of TNF-α,IL-6 and IL-8,the production of ROS,the percentage of apoptotic cells,and the expression of TLR4,p-p65 and IκBα,while increased the expressions of p65 and p-IκBα.In conclusion,borneol alleviates LPS-induced inflammatory response of BMECs by down-regulating TLR4-NF-κB pathway,thus reduce the brain injury.
作者
喻斌
梁涛
卢金福
吕高虹
卞勇
阮鸣
YU Bin;LIANG Tao;LU Jinfu;LYU Gaohong;BIAN Yong;RUAN Ming(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;Laboratory Animal Center,Nanjing University of Chinese Medicine,Nanjing 210023,China;School of Food Science,Nanjing Xiaozhuang University,Nanjing 211117,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2022年第1期1-6,共6页
Immunological Journal
基金
国家自然科学基金(81973726)。
