Equivalence Evaluation of Venenum Bufonis and Its Potential Substitute NJ2196 against LPS-Induced Neuroinflammation in Mice
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摘要:
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, NJ2196 30 mg · kg-1, NJ2196 90 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-α mRNA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。 Abstract: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, NJ2196 30 mg · kg-1 and NJ2196 90 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-1β and TNF-α in mice were detected by ELISA; The mRNA expression levels of inflammatory factors IL-6, TNF-α and 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-1β and TNF-α in mouse serum (P < 0.01), and markedly decreased the expression of inflammatory factors IL-6 and TNF-α mRNA (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. -
Key words:
- Chansu /
- Chansu substitute /
- neuroinflammatory /
- equivalence evaluation
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图 1 蟾酥和NJ2196对小鼠抑郁行为的影响
注: 与对照组比较, ##P < 0.01；与模型组比较, **P < 0.01。x±s, n=9。
Figure 1. Effects of Chansu and NJ2196 on depressive behavior in mice
图 2 蟾酥和NJ2196对小鼠血清炎症因子水平的影响
注: 与对照组比较, ##P < 0.01；与模型组比较, **P < 0.01。x±s, n=6。
Figure 2. Effect of Chansu and NJ2196 on serum inflammatory factor levels in mice
图 3 蟾酥和NJ2196对小鼠海马组织炎症因子mRNA表达的影响
注: 与对照组比较, #P < 0.05，##P < 0.01；与模型组比较, **P < 0.01。x±s, n=5。
Figure 3. Effects of Chansu and NJ2196 on mRNA expression of inflammatory factors in mouse hippocampal tissue
图 4 蟾酥和NJ2196对小鼠海马组织BDNF mRNA表达的影响
注: 与模型组比较, *P < 0.05, **P < 0.01。x±s, n=5。
Figure 4. Effect of Chansu and NJ2196 on BDNF mRNA expression in mouse hippocampal tissue
图 5 蟾酥和NJ2196对脑组织COX途径中炎症介质的影响
注: 与对照组比较, ##P < 0.01;与模型组比较, *P < 0.05, **P < 0.01。x±s, n=5。
Figure 5. Effect of Chansu and NJ2196 on inflammatory mediators in the COX pathway of brain tissue
表 1 引物序列
Table 1. Primer sequences
基因 上游引物(5'→3') 下游引物(5'→3') GAPDH ACCCCAGCAAGGACACTGAGCAAG GGCCCCTCCTGTTAT TATGGGGGT IL-6 AGACAAAGCCAGAGTCCTTCAGAGA GCCACTCCTTCTGTGACTCCAGC TNF-α CCCTCCTGGCCAACGGCATG TCGGGGCAGCCT TGTCCCTT BDNF AGGTCTGACGACGACATCACT CTTCGTTGGGCCGAACCTT 
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[1] 朱伟豪, 吕翔, 沈莉莉, 等. 珍稀动物药代用品研究策略和方法[J]. 中国中药杂志, 2022, 47(17): 612-616. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGZY202217003.htmZHU WH, LYU X, SHEN LL, et al. Research strategies and methods for precious animal medicine substitutes[J]. China J Chin Mater Med, 2022, 47(17): 4560-4564. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGZY202217003.htm [2] HUO YG, XU R, MA HY, et al. Identification of < 10 kD peptides in the water extraction of Venenum Bufonis from Bufo gargarizans using Nano LC-MS/MS and De novo sequencing[J]. J Pharm Biomed Anal, 2018, 157: 156-164. doi: 10.1016/j.jpba.2018.05.027 [3] 王子月, 周婧, 马宏跃, 等. 基于纳升高效液相色谱-线性离子阱-静电场轨道阱高分辨质谱技术研究蟾酥鲜品中的蛋白质[J]. 中国药学杂志, 2017, 52(8): 675-680. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGYX201708012.htmWANG ZY, ZHOU J, MA HY, et al. Identification of proteins in toad venom by NanoLC-LTQ-orbitrap velos pro[J]. Chin Pharm J, 2017, 52(8): 675-680. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGYX201708012.htm [4] WANG JJ, GUO HB, XU DH, et al. Cell affinity screening combined with nanoLC-MS/MS based peptidomics for identifying cancer cell binding peptides from Bufo Bufo gargarizans[J]. J Pharm Biomed Anal, 2021, 206: 114354. doi: 10.1016/j.jpba.2021.114354 [5] XU DH, WANG JJ, CHEN WY, et al. Evaluation of analgesic and anti-inflammatory actions of indolealkylamines from toad venom in mice using lipidomics and molecular docking[J]. J Ethnopharmacol, 2021, 269: 113677. doi: 10.1016/j.jep.2020.113677 [6] 国家药典委员会. 中华人民共和国药典: 一部[M]. 北京: 中国医药科技出版社, 2020: 401.National Pharmacopoeia Committee. Pharmacopoeia of the People's Republic of China: Ⅰ[M]. Beijing: China medical science press, 2020, 401. [7] DANTZER R, O'CONNOR JC, FREUND GG, et al. From inflammation to sickness and depression: When the immune system subjugates the brain[J]. Nat Rev Neurosci, 2008, 9(1): 46-56. doi: 10.1038/nrn2297 [8] HANNESTAD J, DELLAGIOIA N, BLOCH M. The effect of antidepressant medication treatment on serum levels of inflammatory cytokines: A meta-analysis[J]. Neuropsychopharmacology, 2011, 36(12): 2452-2459. doi: 10.1038/npp.2011.132 [9] BRUNING CA, MARTINI F, SOARES SM, et al. Depressive-like behavior induced by tumor necrosis factor-α is attenuated by m-trifluoromethyl-diphenyl diselenide in mice[J]. J Psychiatr Res, 2015, 66/67: 75-83. doi: 10.1016/j.jpsychires.2015.04.019 [10] GE L, LIU LW, LIU HS, et al. Resveratrol abrogates lipopolysaccharide-induced depressive-like behavior, neuroinflammatory response, and CREB/BDNF signaling in mice[J]. Eur J Pharmacol, 2015, 768: 49-57. doi: 10.1016/j.ejphar.2015.10.026 [11] LEE BH, PARK YM, HWANG JA, et al. Variable alterations in plasma erythropoietin and brain-derived neurotrophic factor levels in patients with major depressive disorder with and without a history of suicide attempt[J]. Prog Neuropsychopharmacol Biol Psychiatry, 2021, 110: 110324. doi: 10.1016/j.pnpbp.2021.110324 [12] LUAN SX, ZHOU B, WU Q, et al. Brain-derived neurotrophic factor blood levels after electroconvulsive therapy in patients with major depressive disorder: A systematic review and meta-analysis[J]. Asian J Psychiatr, 2020, 51: 101983. doi: 10.1016/j.ajp.2020.101983 [13] 汪洋, 王玉娟, 武九龙, 等. 电针对PSD大鼠行为学及神经营养因子和炎症因子的影响[J]. 南京中医药大学学报, 2021, 37(2): 263-269. https://www.cnki.com.cn/Article/CJFDTOTAL-NJZY202102020.htmWANG Y, WANG YJ, WU JL, et al. Effect of electroacupuncture on regulation of serum inflammatory factors and nerve cytokines and nerve function in PSD rats[J]. J Nanjing Univ Tradit Chin Med, 2021, 37(2): 263-269. https://www.cnki.com.cn/Article/CJFDTOTAL-NJZY202102020.htm [14] YIN R, ZHAO SF, QIU CX. Brain-derived neurotrophic factor fused with a collagen-binding domain inhibits neuroinflammation and promotes neurological recovery of traumatic brain injury mice via TrkB signalling[J]. J Pharm Pharmacol, 2020, 72(4): 539-550. doi: 10.1111/jphp.13233 [15] ZHU YY, OUYANG ZJ, DU HJ, et al. New opportunities and challenges of natural products research: When target identification meets single-cell multiomics[J/OL]. Acta Pharm Sin B, 2022, 1-29. https://www.sciencedirect.com/science/article/pii/S2211383522003732. -
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