黄岑苷对实验性自身免疫性脑脊髓炎小鼠的影响

目的观察不同浓度黄岑苷(baicalin)对实验性自身免疫脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠模型的作用,并研究其初步机制。方法建立实验性自身免疫性脊髓炎小鼠模型,免疫后第3天给予高低剂量黄岑苷灌胃,每天1次,共20 d。对小鼠活动进行神经功能评分,TUNEL染色检测脊髓组织细胞凋亡情况,ATP水平检测试剂盒检测脊髓组织ATP含量水平,免疫印迹法(Western blot)检测Bax、Bcl-2、cleaved cas-9和cleaved cas-3蛋白表达水平。结果黄岑苷能够提高实验性自身免疫脑脊髓炎小鼠神经功能,延后发病时间;黄岑苷干预后,Tunel染色结果显示脊髓组织凋亡细胞数下降,ATP水平下降,免疫印迹法结果显示Bcl-2的蛋白表达显著上升,Bax、cleaved cas-9和cleaved cas-3的蛋白表达显著下降。结论黄岑苷可通过抑制线粒体内源性凋亡途径抑制细胞凋亡,保护线粒体,提高实验性自身免疫脑脊髓炎小鼠神经功能,为预防疾病提供了实验理论依据。

Protective effect of Baicalin on experimental autoimmune encephalomyelitis in mice

Objective To observe the effects of different concentrations of baicalin on the mouse model of experimental autoimmune encephalomyelitis (EAE) and to explore its mechanisms. Methods A mouse model of EAE was established with MOG33-55 peptide and bacillus Calmette-Guerin (BGG) vaccine with complete Freund adjuvant (CFA). At the third day after immunization, high and low doses of baicalin were administered to the mice intragastrically once a day for 20 days. The neurological function of mice was evaluated. TUNEL staining was used to detect apoptosis in the spinal cord tissue. The level of ATP in spinal cord tissue was detected by an ATP determination kit. Furthermore, the protein expressions of Bax, Bcl-2, cleaved cas-3 and cleaved cas-9 were detected by western blot, respectively. Results Baicalin improved the neurological function and delayed the onset time in EAE mice.After the treatment with baicalin, the TUNEL staining showed that the number of apoptotic cells in spinal cord was decreased, and the ATP level decreased. Western blot revealed that the protein expression of Bcl-2 was significantly increased, while the protein expression of Bax, cleaved cas-3 and cleaved cas-9 were significantly decreased. Conclusions Baicalin can reduce apoptosis by inhibiting mitochondrial endogenous apoptosis pathway, protect the function of mitochondria and improve the neurological function inEAE mice, therefore, provide experimental evidence for the disease prevention.