Stem Cell Secretome Modulated by Arsenicum album 30C Ameliorates Lipopolysaccharide-induced Cytokine Storm in Blood Mononuclear Cells in vitro

IF 1.2 4区医学 Q3 INTEGRATIVE & COMPLEMENTARY MEDICINE

Homeopathy Pub Date : 2023-12-07 DOI:10.1055/s-0043-1776039

Parth Aphale, Avinash Sanap, Dharmendra Sharma, Avinash Kharat, Supriya Kheur, Chinmay Gawade, Indumati Somasundaram, Ramesh Bhonde

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引用次数: 0

Abstract

Background The therapeutic effectiveness of mesenchymal stem cells (MSCs) and their secretome can be enhanced by means of physical, chemical and biological preconditioning. Arsenicum album 30C (AA30) has been one of the leading homeopathic medicines used in prophylaxis against SARS-CoV-2 infection.

Aims This study aimed to investigate whether AA30 preconditioning could influence the growth factors and cytokine profile of the human dental pulp-derived MSC (DPD-MSC) secretome. Also, to test the efficacy of the AA30-preconditioned DPD-MSC secretome in ameliorating the lipopolysaccharide (LPS)-induced cytokine storm in human peripheral blood mononuclear cells (PBMCs) as an in-vitro cellular model.

Methods The cytotoxicity of AA30 was assessed in DPD-MSCs by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Growth factors and cytokine levels in the AA30-preconditioned DPD-MSC secretome were analysed by fluorescence-activated cell sorting (FACS) analysis. The angiogenic potential of the AA30-preconditioned DPD-MSC secretome was assessed by chick yolk-sac membrane (YSM) assay. Culture medium with 0.001% ethanol was used as vehicle control. The efficacy of the AA30-preconditioned DPD-MSC secretome in ameliorating the cytokine storm was assessed in LPS pre-treated PBMCs. The mRNA and protein expression of inflammatory markers such as IL-1β, IL-6 and IL-10 were analysed by using RT-PCR and FACS analysis respectively.

Results AA30 did not exhibit cytotoxicity in the concentration range of 1% to 50%. Furthermore, the AA30-preconditioned DPD-MSC secretome exhibited a significant increase in the levels of angiogenic factors, such as human angiopoietin-2, EPO and PDGF-AA, and decreased levels of cytokines, such as TNF-α, CXCL-8 and IL-6. The AA30-preconditioned DPD-MSC secretome showed augmented angiogenesis compared to vehicle controls. The DPD-MSC secretome ameliorated LPS-induced mRNA and protein expression of IL-1β, IL-6 and IL-10 in PBMCs.

Conclusion The AA30-preconditioned DPD-MSC secretome augmented angiogenesis and ameliorated the LPS-induced cytokine storm in human PBMCs in vitro. Our data demonstrate that AA30 preconditioning enhances the therapeutic potency of MSCs and their secretome.

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砒霜 30C 可调节干细胞分泌组，改善体外血单核细胞中脂多糖诱导的细胞因子风暴

背景间充质干细胞（MSCs）及其分泌组的治疗效果可通过物理、化学和生物预处理得到增强。砒霜30C（AA30）是预防SARS-CoV-2感染的主要顺势疗法药物之一。本研究旨在探讨 AA30 预处理是否能影响人牙髓间充质干细胞（DPD-MSC）分泌组的生长因子和细胞因子谱。此外，还以体外细胞模型为基础，测试经 AA30 预处理的牙髓间充质干细胞分泌组在改善脂多糖（LPS）诱导的人外周血单核细胞（PBMCs）细胞因子风暴方面的功效。方法通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑试验评估 AA30 在 DPD-MSCs 中的细胞毒性。荧光激活细胞分拣（FACS）分析法分析了AA30预处理的DPD-间充质干细胞分泌组中的生长因子和细胞因子水平。AA30预处理的DPD-间充质干细胞分泌组的血管生成潜力通过鸡卵黄囊膜（YSM）试验进行评估。含0.001%乙醇的培养基作为载体对照。在经 LPS 预处理的 PBMCs 中，评估了经 AA30 预处理的 DPD-MSC 分泌组在改善细胞因子风暴方面的功效。采用 RT-PCR 和 FACS 分析方法分别对 IL-1β、IL-6 和 IL-10 等炎症标志物的 mRNA 和蛋白表达进行了分析。结果 AA30 在 1%至 50%的浓度范围内没有细胞毒性。此外，经 AA30 预处理的 DPD-间充质干细胞分泌组的血管生成因子（如人血管生成素-2、EPO 和 PDGF-AA）水平显著增加，细胞因子（如 TNF-α、CXCL-8 和 IL-6）水平下降。与药物对照组相比，AA30 预处理的 DPD-MSC 分泌组显示血管生成增强。DPD-MSC 分泌组能改善 LPS 诱导的 PBMCs 中 IL-1β、IL-6 和 IL-10 的 mRNA 和蛋白表达。结论经 AA30 预处理的 DPD-MSC 分泌体可促进血管生成，并改善体外人 PBMCs 中 LPS 诱导的细胞因子风暴。我们的数据表明，AA30 预处理增强了间充质干细胞及其分泌组的治疗效力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Homeopathy 医学-全科医学与补充医学

CiteScore

3.40

自引率

70.60%

发文量

审稿时长

20.1 weeks

期刊介绍： Homeopathy is an international peer-reviewed journal aimed at improving the fundamental understanding and clinical practice of homeopathy by publishing relevant high-quality original research articles, reviews, and case reports. It also promotes commentary and debate on matters of topical interest in homeopathy.