多酚化合物LM49对角叉菜胶诱导足肿胀小鼠的抗炎作用及机制研究

doi:10.5246/jcps.2025.01.004

中国药学(英文版) ›› 2025, Vol. 34 ›› Issue (1): 41-54.DOI: 10.5246/jcps.2025.01.004

多酚化合物LM49对角叉菜胶诱导足肿胀小鼠的抗炎作用及机制研究

杨帆¹, 李蕊¹, 刘雯婷¹, 孙舰², 赵承孝¹, 冯秀娥¹, 李青山¹^,²^,^*()

^1. 山西医科大学药学院教育部慢性肾病医药基础研究创新中心, 山西太原 030001
^2. 山西中医药大学基于炎性反应的重大疾病创新药物山西省重点实验室, 山西太原 030619

收稿日期:2024-08-12 修回日期:2024-09-20 接受日期:2024-11-07 出版日期:2025-02-20 发布日期:2025-02-20
通讯作者: 李青山

Unveiling the anti-inflammatory effects and mechanisms of LM49 in a carrageenan-induced acute inflammation model

Fan Yang¹, Rui Li¹, Wenting Liu¹, Jian Sun², Chengxiao Zhao¹, Xiue Feng¹, Qingshan Li¹^,²^,^*()

¹ School of Pharmaceutical Science, Medicinal Basic Research Innovation Center of Chronic Kidney Disease, Ministry of Education, Shanxi Medical University, Taiyuan 030001, Shanxi, China
² Shanxi Key Laboratory of Innovative Drug for the Treatment of Serious Diseases Basing on Chronic Inflammation, Shanxi University of Chinese Medicine, Taiyuan 030619, Shanxi, China

Received:2024-08-12 Revised:2024-09-20 Accepted:2024-11-07 Online:2025-02-20 Published:2025-02-20
Contact: Qingshan Li
Supported by:
The National Natural Science Foundation of China (Grant No. 82003770), the National Science and Technology Major Project of China (Grant No. 2018ZX09711001-001-017), and the Key Research and Development Plan of Shanxi Province (Grant No. 202102130501005).

摘要/Abstract

摘要：

本研究旨在评价 2,4ʹ,5ʹ-三羟基-5,2ʹ-二溴二苯甲酮(LM49)对角叉菜胶诱导足肿胀模型小鼠的抗炎作用及潜在机制。小鼠随机分为6组, 采用1%角叉菜胶溶液构建小鼠足肿胀模型。运用病理学、转录组学、流式细胞术、RT-qPCR、Western blot、分子对接等方法, 研究LM49在角叉菜胶诱导的足肿胀模型中的抗炎作用及其机制。结果显示LM49可明显减轻小鼠足部水肿形成, 降低血清促炎因子 IL-1β和TNF-α水平, 提高血清抑炎因子IL-10水平。转录组学分析显示, LM49组中有453个差异表达基因, KEGG和GO分析表明, LM49抑制NF-κB信号通路, 并影响其他一些免疫炎症信号通路。分子对接揭示了LM49在NF-κB信号通路中的8个关键靶点。Western blot结果证实, LM49能抑制小鼠足组织中p65和IκB-α的磷酸化, 以及MYD88和TLR4的表达。本研究结果为进一步研究LM49的抗炎作用及分子机制提供了重要依据。

关键词: 抗炎, 转录组学, 分子对接, 角叉菜胶, 2,4?,5?-三羟基-5,2?-二溴二苯甲酮

Abstract:

This study aimed to investigate the anti-inflammatory properties and underlying molecular mechanisms of 2,4ʹ,5ʹ-trihydroxyl-5,2ʹ-dibromo diphenylmethanone (LM49) using a carrageenan-induced paw edema model in mice, which serves as a well-established model for acute inflammation. Mice were randomly assigned into six groups, and acute inflammation was induced by injecting 1% carrageenan solution into the paw. To elucidate the anti-inflammatory effects and mechanisms of LM49, a comprehensive approach was employed, including pathology, transcriptomics, flow cytometry, RT-qPCR, Western blotting analysis, and molecular docking analysis. The results demonstrated that LM49 exerted a significant protective effect by reducing paw edema and lowering serum levels of pro-inflammatory cytokines IL-1β and TNF-α, while concurrently elevating anti-inflammatory cytokine IL-10 levels. Transcriptomic analysis identified 453 differentially expressed genes in the LM49-treated group. KEGG and GO enrichment analyses indicated that LM49 suppressed the NF-κB signaling pathway and modulated several other immune-inflammatory pathways. Molecular docking studies identified eight key targets of LM49 within the NF-κB signaling pathway. Furthermore, Western blotting analysis confirmed that LM49 inhibited the phosphorylation of p65 and IκB-α and downregulated the expression of MYD88 and TLR4 in mouse paw tissues. These findings provided a foundational understanding of the anti-inflammatory effects and molecular mechanisms of LM49, paving the way for further in-depth studies in this field.

Key words: Anti-inflammation, Transcriptomic, Molecular docking, Carrageenan, 2,4?,5?-Trihydroxyl-5,2?-dibromo diphenylmethanone

Supporting:

杨帆, 李蕊, 刘雯婷, 孙舰, 赵承孝, 冯秀娥, 李青山. 多酚化合物LM49对角叉菜胶诱导足肿胀小鼠的抗炎作用及机制研究[J]. 中国药学(英文版), 2025, 34(1): 41-54.

Fan Yang, Rui Li, Wenting Liu, Jian Sun, Chengxiao Zhao, Xiue Feng, Qingshan Li. Unveiling the anti-inflammatory effects and mechanisms of LM49 in a carrageenan-induced acute inflammation model[J]. Journal of Chinese Pharmaceutical Sciences, 2025, 34(1): 41-54.

图/表 7

Table 1. Primers used for the RT-qPCR study.

Figure 1. Anti-inflammatory capacity of LM49 on Carr-induced paw edema in mice (n = 10 mice per group). (A) Effect of LM49 on Carr-induced mouse paw swelling. (B) Effect of LM49 on Carr-induced mouse paw swelling at 4 h. (C) Pathological observations of paw tissues stained with H&E staining. Inflammatory cell infiltration and ruined hepatic cords (400× magnification). The quantitative data represent mean ± SD (n = 3). #P < 0.05 for model group vs. normal group; *P < 0.05 for LM49 group and indometacin group vs. model group.

Figure 2. Effects of LM49 on lymphocyte subsets in Carr-induced edema. (A) RT-qPCR analysis of mRNA expression of IL-10, IL-1β, TNF-α, and PGE2 in Carr-induced swollen mouse paw tissues. (B) The CD3+CD4+ and CD3+CD8+ subsets of T cells were assessed by flow cytometry. (C) ELISA of protein expression of IL-10, IL-1β and TNF-α levels in mice. The quantitative data represent mean ± SD (n = 6). #P < 0.05 for model group vs. normal group; *P < 0.05 for LM49 group and indometacin group vs. model group.

Figure 3. Expression analysis of DEGs between model and LM49 groups. (A–B) Volcanic map of DEGs among groups. (C–D) Venn diagram of DEGs among groups. (F) Genomes enrichment analysis of up-/down-regulated DEGs. (G) KEGG enrichment analysis of up-/down-regulated DEGs. (E) Heatmap analysis of model and LM49-H group DEGs.

Table 2. LM49 and some protein macromolecular docking box parameters.

Figure 4. Molecular docking analysis. The blue graphic represents the LM49 molecular structure. Purple graphics are protein-activated sites. Green graphics are amino acid residues. The red graphic indicates hydrogen bonding. (A) LM49 docked with related protein molecules in the NF-κB pathway. (B) LM49 docked with related protein molecules in other related pathways.

Figure 5. Effects of LM49 on NF-κB signaling pathway proteins. (A) Expression of p65, p-p65, IκB-α, and p-IκB-α proteins was assessed by Western blotting analysis. (B) The expression of TLR4 and MYD88 proteins was assessed using Western blotting analysis. The quantitative data represent mean ± SD (n = 3). #P < 0.05 for model group vs. normal group; *P < 0.05 for LM49 group and indometacin group vs. model group.

参考文献 34

[1]	Yadav, S.S.; Nair, R.R.; Singh, K. Editorial: Cause or effect: role of inflammation in metabolic disorder. Front. Endocrinol. 2024, 15, 1359605.
[2]	Sikking, M.A.; Stroeks, S.L.V.M.; Marelli-Berg, F.; Heymans, S.R.B.; Ludewig, B.; Verdonschot, J.A.J. Immunomodulation of myocardial fibrosis. JACC Basic Transl. Sci. 2023, 8, 1477–1488.
[3]	Zhao, R.M.; Wang, B.X.; Wang, D.S.; Wu, B.H.; Ji, P.Y.; Tan, D.Y. Oxyberberine prevented lipopolysaccharide-induced acute lung injury through inhibition of mitophagy. Oxid. Med. Cell Longev. 2021, 2021, 6675264.
[4]	Zhang, X.R.; Retyunskiy, V.; Qiao, S.; Zhao, Y.; Tzeng, C.M. Alloferon-1 ameliorates acute inflammatory responses in λ-carrageenan-induced paw edema in mice. Sci. Rep. 2022, 12, 16689.
[5]	Gonzalez, A.L.; Dungan, M.M.; Smart, C.D.; Madhur, M.S.; Doran, A.C. Inflammation resolution in the cardiovascular system: arterial hypertension, atherosclerosis, and ischemic heart disease. Antioxid. Redox Signal. 2024, 40, 292–316.
[6]	Koenig, W.; Sager, H.B. Inflammation and cardiovascular disease: new epidemiologic data and their potential implications for anti-cytokine therapy. Eur. J. Prev. Cardiol. 2023, 30, 1728–1730.
[7]	Guo, X.Y.; Shen, P.M.; Shao, R.J.; Hong, T.; Liu, W.Z.; Shen, Y.; Su, F.; Wang, Q.L.; He, B. Efferocytosis-inspired nanodrug treats sepsis by alleviating inflammation and secondary immunosuppression. Biomed. Mater. 2023, 18, 055020.
[8]	Feng, Y.Y.; Wang, Z.J.; Pang, H. Role of metformin in inflammation. Mol. Biol. Rep. 2023, 50, 789–798.
[9]	Dai, C.L.; Yang, H.X.; Liu, Q.P.; Rahman, K.; Zhang, H. CXCL6: a potential therapeutic target for inflammation and cancer. Clin. Exp. Med. 2023, 23, 4413–4427.
[10]	Monteleone, G.; Moscardelli, A.; Colella, A.; Marafini, I.; Salvatori, S. Immune-mediated inflammatory diseases: common and different pathogenic and clinical features. Autoimmun. Rev. 2023, 22, 103410.
[11]	Bai, J.J.; Qian, B.L.; Cai, T.Y.; Chen, Y.F.; Li, T.X.; Cheng, Y.L.; Wu, Z.M.; Liu, C.; Ye, M.X.; Du, Y.C.; Fu, W.G. Aloin attenuates oxidative stress, inflammation, and CCl₄-induced liver fibrosis in mice: possible role of TGF-β/smad signaling. J. Agric. Food Chem. 2023, 71, 19475–19487.
[12]	Liu, Z.W.; Gao, T.H.; Yang, Y.; Meng, F.X.; Zhan, F.P.; Jiang, Q.C.; Sun, X. Anti-cancer activity of porphyran and carrageenan from red seaweeds. Molecules. 2019, 24, 4286.
[13]	Borsani, B.; De Santis, R.; Perico, V.; Penagini, F.; Pendezza, E.; Dilillo, D.; Bosetti, A.; Zuccotti, G.V.; D’Auria, E. The role of carrageenan in inflammatory bowel diseases and allergic reactions: where do we stand? Nutrients. 2021, 13, 3402.
[14]	Xiang, L.; Huang, Q.W.; Chen, T.; He, Q.M.; Yao, H.; Gao, Y.X. Ethanol extract of Paridis rhizoma attenuates carrageenan-induced paw swelling in rats by inhibiting the production of inflammatory factors. BMC Complement. Med. Ther. 2023, 23, 437.
[15]	Almukainzi, M.; El-Masry, T.A.; Selim, H.; Saleh, A.; El-Sheekh, M.; Makhlof, M.E.M.; El-Bouseary, M.M. New insight on the cytoprotective/antioxidant pathway Keap1/Nrf2/HO-1 modulation by Ulva intestinalis extract and its selenium nanoparticles in rats with carrageenan-induced paw edema. Mar. Drugs. 2023, 21, 459.
[16]	Salem, S.; Leghouchi, E.; Soulimani, R.; Bouayed, J. Reduction of paw edema and liver oxidative stress in carrageenan-induced acute inflammation by Lobaria pulmonaria and Parmelia caperata, lichen species, in mice. Int. J. Vitam. Nutr. Res. 2021, 91, 143–151.
[17]	Annamalai, P.; Thangam, E.B. Vitex trifolia L. modulates inflammatory mediators via down-regulation of the NF-κB signaling pathway in carrageenan-induced acute inflammation in experimental rats. J. Ethnopharmacol. 2022, 298, 115583.
[18]	Zhang, Q.; Bang, S.S.; Chandra, S.; Ji, R.R. Inflammation and infection in pain and the role of GPR37. Int. J. Mol. Sci. 2022, 23, 14426.
[19]	Khan, M.M.; Ali, S.A.; Qazi, Y.; Khan, S.W.; Shaikh, M.A. Anti-inflammatory effects of Chrozophora plicata uncovered using network pharmacology and in-vivo carrageenan paw edema model. Heliyon. 2024, 10, e24617.
[20]	Lukova, P.; Apostolova, E.; Baldzhieva, A.; Murdjeva, M.; Kokova, V. Fucoidan from Ericaria crinita alleviates inflammation in rat paw edema, downregulates pro-inflammatory cytokine levels, and shows antioxidant activity. Biomedicines. 2023, 11, 2511.
[21]	El Awady, M.E.; Mohamed, S.S.; Abo Elsoud, M.M.; Mahmoud, M.G.; Anwar, M.M.; Ahmed, M.M.; Eltaher, A.; Magdeldin, S.; Attallah, A.; Elhagry, A.E.; Abdelhamid, S.A. Insight into antioxidant and anti-inflammatory effects of marine bacterial natural exopolysaccharide (EPSSM) using carrageenan-induced paw edema in rats. Sci. Rep. 2024, 14, 5113.
[22]	Han, S.; Li, S.Y.; Li, J.L.; He, J.; Wang, Q.Q.; Gao, X.; Yang, S.L.; Li, J.J.; Yuan, R.; Zhong, G.Y.; Gao, H.W. Hederasaponin C inhibits LPS-induced acute kidney injury in mice by targeting TLR4 and regulating the PIP2/NF-κB/NLRP3 signaling pathway. Phytother. Res. 2023, 37, 5974–5990.
[23]	Li, R.; Jia, H.Y.; Si, M.; Li, X.W.; Ma, Z.; Zhu, Y.; Sun, W.Y.; Zhu, F.Q.; Luo, S.Y. Loureirin B protects against cerebral ischemia/reperfusion injury through modulating M1/M2 microglial polarization via STAT6/NF-kappaB signaling pathway. Eur. J. Pharmacol. 2023, 953, 175860.
[24]	Wang, F.; Li, R.; Wang, W,; Zhou, X.; Liu, M.; Zhao, J.; Wen, A.; Wang, W.; Jia, Y. α-Boswellic acid ameliorates acute kidney injury by inhibiting the TLR4-mediated inflammatory pathway. J. Chin. Pharm. Sci. 2023, 32. 539–550.
[25]	Yang, F.; Cai, H.H.; Feng, X.E.; Li, Q.S. A novel marine halophenol derivative attenuates lipopolysaccharide-induced inflammation in RAW_264.7 cells via activating phosphoinositide 3-kinase/Akt pathway. Pharmacol. Rep. 2020, 72, 1021–1031.
[26]	Yang, F.; Cai, H.H.; Zhang, X.; Sun, J.; Feng, X.E.; Yuan, H.X.; Zhang, X.Y.; Xiao, B.G.; Li, Q.S. An active marine halophenol derivative attenuates lipopolysaccharide-induced acute liver injury in mice by improving M2 macrophage-mediated therapy. Int. Immunopharmacol. 2021, 96, 107676.
[27]	Sen’kova, A.V.; Savin, I.A.; Odarenko, K.V.; Salomatina, O.V.; Salakhutdinov, N.F.; Zenkova, M.A.; Markov, A.V. Protective effect of soloxolone derivatives in carrageenan- and LPS-driven acute inflammation: pharmacological profiling and their effects on key inflammation-related processes. Biomed. Pharmacother. 2023, 159, 114231.
[28]	Aitbaba, A.; Kabdy, H.; Fatimzahra, A.; Azraida, H.; Abdoussadeq, O.; Aboufatima, R.; El Yazouli, L.; Sokar, Z.; Garzoli, S.; Chait, A. Anti-inflammatory and antiarthritic properties of Marrubium vulgare aqueous extract in an animal model. Nat. Prod. Res. 2024, 1–9.
[29]	Zhou, X.; Guo, Z.L.; Liu, S.Z.; Chen, Z.J.; Wang, Y.; Yang, R.; Li, X.Z.; Ma, K.T. Transcriptomics and molecular docking reveal the potential mechanism of lycorine against pancreatic cancer. Phytomedicine. 2024, 122, 155128.
[30]	Xu, X.; Li, L.; Zhang, Y.; Lu, X.; Xu, R.; Wu, S.; Lin, W.; Hypolipidemic effects of Alismatis Rhizoma decoction on the lipid profile in hyperlipidemia rats by RNA-sequencing. J. Chin. Pharm. Sci. 2021, 30, 409–420.
[31]	Sun, Z.C.; Wang, Y.Q.; Pang, X.Y.; Wang, X.Y.; Zeng, H. Mechanisms of polydatin against spinal cord ischemia-reperfusion injury based on network pharmacology, molecular docking and molecular dynamics simulation. Bioorg. Chem. 2023, 140, 106840.
[32]	Agu, P.C.; Afiukwa, C.A.; Orji, O.U.; Ezeh, E.M.; Ofoke, I.H.; Ogbu, C.O.; Ugwuja, E.I.; Aja, P.M. Molecular docking as a tool for the discovery of molecular targets of nutraceuticals in diseases management. Sci. Rep. 2023, 13, 13398.
[33]	Kumar, A.; Singh, V.K.; Kayastha, A.M. Molecular modeling, docking and dynamics studies of fenugreek (Trigonella foenum-graecum) α-amylase. J. Biomol. Struct. Dyn. 2023, 41, 9297–9312.
[34]	Bisht, A.; Tewari, D.; Kumar, S.; Chandra, S. Network pharmacology, molecular docking, and molecular dynamics simulation to elucidate the mechanism of anti-aging action of Tinospora cordifolia. Mol. Divers. 2024, 28, 1743–1763.

多酚化合物LM49对角叉菜胶诱导足肿胀小鼠的抗炎作用及机制研究

Unveiling the anti-inflammatory effects and mechanisms of LM49 in a carrageenan-induced acute inflammation model

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