蟾酥及其潜在代用品NJ2196抗LPS诱导小鼠神经炎症的等效性评价

Equivalence Evaluation of Venenum Bufonis and Its Potential Substitute NJ2196 against LPS-Induced Neuroinflammation in Mice

目的评价蟾酥及其潜在代用品NJ2196抗脂多糖(LPS)诱导小鼠神经炎症的等效性。方法63只SPF级雄性ICR小鼠随机分为对照组、模型组、氟西汀组、蟾酥低剂量组、蟾酥高剂量组、NJ2196低剂量组、NJ2196高剂量组,每组9只。除对照组外,每组单次给予LPS(0.83 mg·kg^(-1),腹腔注射)建立炎性抑郁模型。氟西汀组、蟾酥低剂量组、蟾酥高剂量组、NJ2196低剂量组、NJ2196高剂量组分别灌胃氟西汀30 mg·kg^(-1),蟾酥30 mg·kg^(-1),蟾酥90 mg·kg^(-1),NJ219630 mg·kg^(-1),NJ219690 mg·kg^(-1),对照组和模型组给予等量生理盐水。观察小鼠悬尾累积不动时间和强迫游泳累积不动时间;ELISA法检测小鼠血清炎症因子白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α水平;qPCR法检测海马组织炎症因子IL-6和TNF-α以及脑源性神经营养因子(BDNF)mRNA表达水平;高灵敏度脂质组学分析蟾酥和NJ2196对炎性类花生酸的影响。结果与模型组相比,蟾酥和NJ2196能够显著降低小鼠悬尾累积不动时间(P<0.01)和强迫游泳累积不动时间(P<0.01),显著降低小鼠血清中炎症因子IL-1β、TNF-α水平(P<0.01),显著降低小鼠海马体中炎症因子IL-6、TNF-αmRNA表达水平(P<0.01)以及BDNF mRNA表达水平(P<0.01),高灵敏度脂质组学分析显示,炎症因子的变化可能与来自环氧合酶(COX)途径的炎症介质的下调有关。各剂量蟾酥与NJ2196比较无显著性差异(P>0.05)。结论蟾酥及其潜在代用品NJ2196具有显著的抗炎性抑郁活性,在LPS诱导小鼠神经炎症模型上两者具有一定等效性。

OBJECTIVE To evaluate the equivalence between Venenum Bufonis(Chansu)and its potential substitute NJ2196 against lipopolysaccharide(LPS)-induced neuroinflammation in mice.METHODS Sixty-three SPF male ICR mice were randomly divided into control group,model group,fluoxetine group,toadstool low-dose group,toadstool high-dose group,NJ2196 low-dose group,and NJ2196 high-dose group,with 9 mice in each group.A single administration of LPS(0.83 mg·kg^(-1),intraperitoneal injection)was given to each group except the control group to establish an inflammatory depression model.Fluoxetine group,Chansu low dose group,Chansu high dose group,NJ2196 low dose group and NJ2196 high dose group were given fluoxetine 30 mg·kg^(-1),Chansu 30 mg·kg^(-1),Chansu 90 mg·kg^(-1),NJ219630 mg·kg^(-1)and NJ219690 mg·kg^(-1)by gavage,respectively,and the control and model group was given equal amount of saline.The cumulative immobility time of tail suspension and the cumulative immobility time of forced swimming in mice were observed;The levels of serum inflammatory factors IL-1βand TNF-αin mice were detected by ELISA;The mRNA expression levels of inflammatory factors IL-6,TNF-αand brain-derived neurotrophic factor(BDNF)in hippocampal tissues were detected by qPCR;The effects of Chansu and NJ2196 on inflammatory arachidonic acids were analyzed by high-sensitivity lipidomics.RESULTS Compared with the model group,Chansu and NJ2196 significantly reduced the cumulative immobility time of tail suspension(P<0.01)and the cumulative immobility time of forced swimming(P<0.01),both significantly reduced the levels of inflammatory factors IL-1βand TNF-αin mouse serum(P<0.01),and markedly decreased the expression of inflammatory factors IL-6 and TNF-αmRNA(P<0.01)as well as BDNF mRNA expression(P<0.01)in mouse hippocampus.High-sensitivity lipidomic analysis showed that the changes in inflammatory factors might be related to the downregulation of inflammatory mediators from the cyclooxygenase(COX)pathway.There was no significant difference(P>0.05)between Chansu and NJ2196 in all dose groups.CONCLUSION Chansu and its potential substitute NJ2196 have significant anti-inflammatory depressive activities,and also have equivalence in the LPS-induced neuroinflammation model.