Sun Ke, Luo Zhao-Liang, Hu Chen, Gong Ting-Liang, Tang Guo-Hua, Wu Shao-Ping
Department of Brain Disease, Chongqing 400011, China.
Department of Cardiology, Chongqing Traditional Chinese Medicine Hospital, Chongqing 400011, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Nov;36(6):656-661. doi: 10.12047/j.cjap.6001.2020.137.
To investigate the protective effect and immune mechanism of berberine on cerebral ischemia/reperfusion injury in rats. Fifty SD rats were randomly divided into sham operation group (Sham), model group (Model), berberine low dose groups (BBR-L, 25 mg/kg), berberine medium dose groups (BBR-M, 50 mg/kg) and berberine high dose groups (BBR-H, 100 mg/kg), with 10 rats in each group. Longa suture method was used to establish a rat model of cerebral ischemia/reperfusion, after 2 hours of ischemia, reperfusion for 24 hours. Rats in BBR-L, BBR-M and BBR-H were treated with berbrerine by gavage 2 hours after successful model building, while the sham operation group and the modle group were given the same volume of saline as described above. After 24 hours of administration, the activity of antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), cytokine tumor necrosis factor α (TNF-α) , interferon β (IFN-β) , interleukin 6 (IL-6) and nitric oxide (NO) content were detected by ELISA assay. Serum CD4, CD8 and CD4/CD8 contents were measured by flow cytometry to investigate the immune function of each group. RT-qPCR and Western blot were used to detect NF-kappaB-NOD-like receptors 3 (NF-κB-NLRP3) signal axis key genes and protein expression in rat brain tissue. Compared with the sham operation group, the degree of neurological deficit and the rate of cerebral infarction were increased in the model group (＜0.05), and the levels of serum NO, TNF-α, IFN-β, IL-6, NF-κB p65, NLRP3, ASC and caspase-1 in brain tissue were increased (＜0.05), while the activities of SOD, GSH-Px and the levels of CD4, CD8 and CD4/CD8 in serum were decreased (＜0.05). Compared with the model group, the degree of neurological deficit and the rate of cerebral infarction were increased in the BBR-H, BBR-M and BBR-L groups (＜0.05), and the levels of serum NO, TNF-α, IFN-β, IL-6, NF-κB p65, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) and caspase-1 in brain tissue were increased (＜0.05), while the activities of SOD, GSH-Px and the levels of CD4, CD8 and CD4/CD8 in serum were decreased (＜0.05). Berberine may reduce oxidative stress, inhibit inflammation, enhance immune function, and reduce cerebral ischemia/reperfusion injury in rats, which may be related to the inhibition of NF-κB-NLRP3 signaling.
探讨黄连素对大鼠脑缺血/再灌注损伤的保护作用及免疫机制。将50只SD大鼠随机分为假手术组（Sham）、模型组（Model）、黄连素低剂量组（BBR-L，25mg/kg）、黄连素中剂量组（BBR-M，50mg/kg）和黄连素高剂量组（BBR-H，100mg/kg），每组10只。采用Longa线栓法建立大鼠脑缺血/再灌注模型，缺血2小时后，再灌注24小时。BBR-L、BBR-M和BBR-H组大鼠在模型成功建立后2小时经灌胃给予黄连素，假手术组和模型组给予与上述相同体积的生理盐水。给药24小时后，采用ELISA法检测抗氧化酶超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）的活性、细胞因子肿瘤坏死因子α（TNF-α）、干扰素β（IFN-β）、白细胞介素6（IL-6）及一氧化氮（NO）含量。采用流式细胞术检测血清CD4、CD8及CD4/CD8含量，以探讨各组的免疫功能。采用RT-qPCR和Western blot法检测大鼠脑组织中核因子κB-核苷酸结合寡聚化结构域样受体3（NF-κB-NLRP3）信号轴关键基因及蛋白表达。与假手术组比较，模型组神经功能缺损程度及脑梗死率升高（P＜0.05），血清NO、TNF-α、IFN-β、IL-6、脑组织中NF-κB p65、NLRP3、凋亡相关斑点样蛋白（ASC）及半胱天冬酶-1水平升高（P＜0.05），而血清SOD、GSH-Px活性及CD4、CD8、CD4/CD8水平降低（P＜0.05）。与模型组比较，BBR-H、BBR-M和BBR-L组神经功能缺损程度及脑梗死率降低（P＜0.05），血清NO、TNF-α、IFN-β、IL-6、脑组织中NF-κB p65、NLRP3、ASC及半胱天冬酶-1水平降低（P＜0.05），而血清SOD、GSH-Px活性及CD4、CD8、CD4/CD8水平升高（P＜0.05）。黄连素可能通过抑制NF-κB-NLRP3信号通路减轻氧化应激、抑制炎症反应、增强免疫功能，从而减轻大鼠脑缺血/再灌注损伤。
