Chen Hai-Dong, Jiang Ming-Zhao, Zhao Ying-Ying, Li Xin, Lan Hai, Yang Wan-Qi, Lai Yong
College of Pharmacy, Dali University, Dali, 671000, Yunnan, People's Republic of China; Yunnan Key Laboratory of Screening and Research on Anti-pathogenic Plant Resources from Western Yunnan (Cultivation), Dali, People's Republic of China.
College of Pharmacy, Dali University, Dali, 671000, Yunnan, People's Republic of China.
J Ethnopharmacol. 2023 Jan 10;300:115691. doi: 10.1016/j.jep.2022.115691. Epub 2022 Sep 7.
The plant Erigeron breviscapus (Vant.) Hand.-Mazz.,a Chinese herbal medicine with multiple pharmacological effects and clinical applications, has been traditionally used in the treatment of paralysis caused by stroke and joint pain from rheumatism by the Yi minority people of Southwest China for generations.However, its mechanism involves many factors and has not been fully clarified.
Taking intestinal flora as the target, the protective effect of extract(breviscapine) of E. breviscapus on cerebral ischemia and its possible mechanism were discussed from the perspective of brain inflammatory pathway and intestinal CYP3A4, which depends on intestinal flora.
In this study, we first verified the binding ability between major active ingredient of Erigeron breviscapus and the core target TLR4 protein by molecular docking using Vina software.We established a rat model of cerebral ischemia-reperfusion injury in vivo.The neurological function of rats was scored by Bederson score table, the cerebral infarction volume was detected by TTC staining, and the serum NSE level was detected by ELASA. 16S rRNA sequencing was used to detect the intestinal flora of rats in each group.The expression levels of cerebral TLR4/MyD88/NF-κB and CYP3A4 mRNA and protein in different intestinal segments were detected by qRT-PCR and Western blot.
Compared with the model group, the neurological injury score, infarct volume and serum NSE concentration of breviscapine low, medium and high dose groups and nimodipine groups decreased significantly. Meanwhile, breviscapine could significantly reduce the expression level of the TLR4/MyD88/NF-κB in brain tissue and CYP3A4 in different intestinal segments of rats with cerebral ischemia-reperfusion injury. In addition, breviscapine also significantly ameliorated intestinal flora dysbiosis of rats with cerebral ischemia-reperfusion injury.
Breviscapine can protect rats from cerebral ischemia-reperfusion injury by regulating intestinal flora, inhibiting brain TLR4/MyD88/NF-κB inflammatory pathway and intestinal CYP3A4 expression.
灯盏花（Erigeron breviscapus (Vant.) Hand.-Mazz.）是一种具有多种药理作用和临床应用的中药材，在中国西南部彝族中，世代相传用于治疗中风引起的瘫痪和风湿性关节疼痛。然而，其作用机制涉及多种因素，尚未完全阐明。
以肠道菌群为靶点，从脑炎症通路和依赖肠道菌群的肠道CYP3A4角度，探讨灯盏花提取物（灯盏花素）对脑缺血的保护作用及其可能机制。
本研究首先使用Vina软件通过分子对接验证灯盏花主要活性成分与核心靶点TLR4蛋白的结合能力。我们建立了大鼠脑缺血再灌注损伤体内模型。采用Bederson评分表对大鼠神经功能进行评分，通过TTC染色检测脑梗死体积，采用酶联免疫吸附测定法（ELASA）检测血清神经元特异性烯醇化酶（NSE）水平。使用16S rRNA测序检测各组大鼠的肠道菌群。采用实时定量聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法（Western blot）检测不同肠段脑TLR4/髓样分化因子88（MyD88）/核因子κB（NF-κB）和CYP3A4 mRNA及蛋白的表达水平。
与模型组相比，灯盏花素低、中、高剂量组和尼莫地平组的神经损伤评分、梗死体积和血清NSE浓度均显著降低。同时，灯盏花素可显著降低脑缺血再灌注损伤大鼠脑组织中TLR4/MyD88/NF-κB和不同肠段CYP3A4的表达水平。此外，灯盏花素还显著改善了脑缺血再灌注损伤大鼠的肠道菌群失调。
灯盏花素可通过调节肠道菌群、抑制脑TLR4/MyD88/NF-κB炎症通路和肠道CYP3A4表达来保护大鼠免受脑缺血再灌注损伤。
