中药新药与临床药理2025，Vol.36Issue(4)：522-533,12.DOI:10.19378/j.issn.1003-9783.2025.04.005
青蒿琥酯联合血必净注射液通过VEGF/Akt/eNOS通路对实验性脑型疟小鼠的保护作用及机制探讨
Protective Effect and Mechanism of Artesunate Combined with Xuebijing Injection on Experimental Cerebral Malaria Mice Through VEGF/Akt/eNOS Pathway
摘要
Abstract
Objective To investigate the protective effect and mechanism of artesunate combined with Xuebijing Injection on experimental cerebral malaria(ECM)mice.Methods Female C57BL/6 mice aged 4-5 weeks were randomly divided into blank control group(0.9%NaCl),infection control group(0.9%NaCl),Xuebijing Injection group(Xuebijing,8 mL·kg-1,5 days),artesunate group(artesunate,20 mg·kg-1,double the first dose,5 days),artesunate combined with Xuebijing Injection group("combination group").Xuebijing 8 mL·kg-1+artesunate 20 mg·kg-1,double the first dose of artesunate,5 days),20 rats in each group.According to the above groups,intraperitoneal injection was given once a day.The body mass,body temperature,peripheral blood infection rate and survival of mice were recorded.Evans blue staining was used to detect the permeability of blood-brain barrier in mouse brain tissue;the pathological changes of brain tissue in mice were detected by hematoxylin-eosin(HE)staining.The levels of tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ)in serum of mice were detected by ELISA.Western Blot was used to detect the protein levels of Occludin,ZO-1,VE-Cadherin,Ang-1,Ang-2,ICAM-1,VCAM-1,VEGF,PI3 K,Akt,p-Akt,eNOS and p-eNOS in brain tissue of mice.The level of ICAM-1 protein in brain tissue of mice was detected by immunofluorescence.The level of nitric oxide(NO)in brain tissue of mice was detected by total nitric oxide detection kit.Results Compared with the blank control group,the body temperature and body mass of the mice in the infection control group decreased(P＜0.01).At the end of administration,the infection rate reached 30.99%,and the survival rate was 0.The mice showed neurological symptoms such as limp and loss of reflex.The Rapid Murine Coma and Behavioral Scale(RMCBS)score decreased(P＜0.01),and the permeability of the blood-brain barrier increased(P＜0.05).There were infected red blood cells and white blood cells adhered to the brain microvessels,and there were bleeding points.The levels of TNF-α and IFN-γ in serum increased(P＜0.05,P＜0.01).The protein expressions of Occludin,ZO-1,VE-Cadherin,Ang-1 and p-Akt/Akt in brain tissue decreased(P＜0.05,P＜0.01),the protein expressions of ICAM-1 and VCAM-1 increased(P＜0.05,P＜0.01),and the expression of NO decreased(P＞0.05);compared with the infection control group,the body temperature and body mass of the mice in the combined treatment group increased(P＜0.01).At the end of administration,the infection rate was 0.67%,the inhibition rate reached 97.84%,the survival rate was 20%,the neurological symptoms of the mice were improved,the RMCBS score increased(P＜0.01),the permeability of the blood-brain barrier decreased(P＞0.05),there was no obvious cell adhesion in the brain microvessels,and the levels of TNF-α and IFN-γ in the serum decreased(P＜0.05,P＜0.01).The protein expressions of Occludin,ZO-1,VE-Cadherin,Ang-1,VEGF,PI3K,p-Akt/Akt and p-eNOS/eNOS in brain tissue increased(P＜0.05,P＜0.01),the expressions of ICAM-1 and VCAM-1 decreased(P＜0.01),and the expression of NO increased(P＜0.01).Conclusion Artesunate combined with Xuebijing Injection can effectively improve the symptoms and signs of ECM mice,reduce the adhesion in brain microvessels,alleviate excessive inflammatory reaction and reduce the permeability of blood-brain barrier.The mechanism may be related to the activation of VEGF/Akt/eNOS signaling pathway and the increase of NO expression.关键词
脑型疟/青蒿琥酯/血必净注射液/VEGF/Akt/eNOS信号通路/NO/小鼠Key words
cerebral malaria/artesunate/Xuebijing Injection/VEGF/Akt/eNOS signaling pathway/NO/mice分类
医药卫生引用本文复制引用
陈培婷,谭瑞湘,邱翠雯,余林辛,周玖瑶,邓长生,宋健平..青蒿琥酯联合血必净注射液通过VEGF/Akt/eNOS通路对实验性脑型疟小鼠的保护作用及机制探讨[J].中药新药与临床药理,2025,36(4):522-533,12.基金项目
国家中医药管理局中医药国际合作专项(XDZYJZC-001) （XDZYJZC-001）
国家自然科学基金项目(82374315). （82374315）
