DataSheet4_The Protective Effect of Pilose Antler Peptide on CUMS-Induced Depression Through AMPK/Sirt1/NF-κB/NLRP3-Mediated Pyroptosis.docx

Background: Pilose antler peptide (PAP), prepared from the pilose antler of Cervus nippon Temminck, is widely used in traditional Chinese medicine (TCM) against various inflammatory disorders. TCM prescriptions containing pilose antler are often prescribed clinically to treat depression. However, the pharmacological mechanisms of how PAP, against inflammation, prevents and treats depression remain poorly understood.
Methods: PAP was identified by de novo sequencing and database searching. Then, behavioral tests were conducted to investigate the effect of PAP on CUMS-exposed mice. In parallel, Nissl staining and Golgi-Cox staining were used for exploring the effect of PAP on neural cells and dendritic spine density. Additionally, the expression of key proteins of the AMPK/Sirt1/NF-κB/NLRP3 pathway was analyzed by Western blot. Finally, the CUMS procedure was conducted for 6 weeks. At the 5th week, PAP and fluoxetine (Flu) were intragastrically treated for 2 weeks. The silencing information regulator-related enzyme 1 (Sirt1) inhibitor EX-527 and the AMP-activated protein kinase (AMPK) inhibitor dorsomorphin were employed to investigate the effects of Sirt1 and AMPK on PAP-mediated depression.
Results: PAP attenuated the behavior alteration caused by CUMS stimulation, decreased the number of neurons, and restored the dendritic spine density. PAP treatment effectively upregulated the expressions of p-AMPK and Sirt1 and suppressed the expressions of Ac-NF-κB, NLRP3, Ac-Caspase-1, GSDMD-N, Cleaved-IL-1β, and Cleaved-IL-18. Moreover, selectively inhibited Sirt1 and AMPK were able to compromise the therapeutic effect of PAP on depression.
Conclusion: The present work indicated that PAP has a protective effect on CUMS-induced depression. In addition, AMPK and Sirt1 played critical roles in the PAP-relieved depression. PAP might be a potential therapeutic option for treating depression.

背景：从梅花鹿（Cervus nippon Temminck）鹿茸中提取制备的鹿茸肽（Pilose antler peptide, PAP），在传统中医药（Traditional Chinese Medicine, TCM）中被广泛用于治疗多种炎症性疾病。含鹿茸的中药复方在临床中常被用于治疗抑郁症，但鹿茸肽通过抗炎作用预防和治疗抑郁症的具体药理机制仍尚不明确。
方法：本研究通过从头测序与数据库检索鉴定得到鹿茸肽。随后采用行为学实验探究鹿茸肽对慢性不可预知温和应激（Chronic Unpredictable Mild Stress, CUMS）造模小鼠的干预作用。同时，利用尼氏染色与高尔基-科克斯染色，观察鹿茸肽对神经细胞及树突棘密度的影响。此外，通过蛋白质印迹法（Western blot）分析腺苷酸活化蛋白激酶/沉默信息调节因子1/核因子κB/NLRP3通路关键蛋白的表达水平。本研究全程采用CUMS造模6周，于造模第5周开始，通过灌胃给予鹿茸肽与氟西汀（Flu），持续给药2周。为探究沉默信息调节因子1（Sirt1）与腺苷酸活化蛋白激酶（AMPK）在鹿茸肽改善抑郁症中的调控作用，本研究分别采用Sirt1抑制剂EX-527与AMPK抑制剂dorsomorphin开展干预实验。
结果：鹿茸肽可缓解CUMS刺激诱导的行为学异常，改善神经元数量减少并恢复树突棘密度。鹿茸肽给药可有效上调p-AMPK与Sirt1的表达水平，并下调Ac-NF-κB、NLRP3、Ac-Caspase-1、GSDMD-N、Cleaved-IL-1β及Cleaved-IL-18的表达。此外，抑制Sirt1或AMPK的活性可削弱鹿茸肽对抑郁症的治疗效果。
结论：本研究证实鹿茸肽对CUMS诱导的抑郁症具有明确保护作用。腺苷酸活化蛋白激酶（AMPK）与沉默信息调节因子1（Sirt1）在鹿茸肽缓解抑郁症的过程中发挥关键调控作用。鹿茸肽有望成为治疗抑郁症的潜在候选治疗方案。