尿石素C通过阻断脂多糖诱导的RAW 264.7巨噬细胞中的NF-κB信号通路来抑制炎症。

Urolithin-C Suppresses Inflammation by Blocking NF-κB Signaling Pathway in LPS-Induced RAW 264.7 Macrophages.

作者信息

Manjappa Vani Karadi, Venkatappa Manjula Mavathuru, Urs Deepadarshan, Basavarajaiah Suliphuldevara Mathada, Venkataramaiah Shivakumar, Mahendranathsingh Sanjana Bai Shivramsingh, Mohan Sushma, Pushpavathi Hunase Rajaiah, Krishnappa Dharmappa Kattepura, Sannaningaiah Devaraja

机构信息

Department of Studies and Research in Biochemistry, Tumkur University, Tumkur, Karnataka, 572103, India.

Department of Biochemistry, Maharani Lakshmi Ammanni College for Women, Autonomous, Bangalore, Karnataka, 560012, India.

出版信息

J Inflamm Res. 2025 Sep 9;18:12463-12483. doi: 10.2147/JIR.S539273. eCollection 2025.

DOI:10.2147/JIR.S539273

PMID:40951448

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12433252/

Abstract

PURPOSE

In the current study, the evaluation of anti-inflammatory ) activity of chemically synthesized Urolithin-C was examined.

METHODS

The synthesis of Urolithin-C (3,8,9-trihydroxy-6H-benzo[c]chromen-6-one) was carried out by chemical method and it was characterized using various techniques. The anti-inflammatory efficacy of synthesized Urolithin-C was studied by membrane stabilization, protein denaturation and protease inhibition assays. In addition, MTT (3-[4,5-dimethylthiazol-2-yl] 2,5-diphenyl tetrazolium bromide) assay was employed to evaluate the cytotoxic effect of Urolithin-C. The anti-inflammatory property of Urolithin-C was further examined using LPS (Lipopolysaccharide) induced RAW 264.7 (Mouse macrophage) cells. Furthermore, the anti-inflammatory properties of Urolithin-C was studied by quantifying pro/anti-inflammatory cytokines using ELISA (enzyme-linked immunosorbent assay). The mechanism of action of Urolithin-C on NF-κB (Nuclear Factor-kappa B) translocation was studied using CLSM (confocal laser scanning microscopy). While gene expression pattern was analyzed using RT-qPCR (Reverse Transcription quantitative Polymerase Chain Reaction).

RESULTS

In comparison to the positive control aspirin, Urolithin-C showed a strong anti-inflammatory effect by preventing lysosomal degradation, protein denaturation and inhibition of protease. Furthermore, at the higher dose (200 µg/mL), Urolithin-C was found to be toxic to the mouse macrophages; however, at lower concentration (25 µg/mL) it did not cause toxicity to the said. Thus, 25 µg/mL of Urolithin-C was used to assess the anti-inflammatory activity. Interestingly, Urolithin-C efficiently reduced the expression of pro-inflammatory inducible enzyme (Cox-2), cytokines (IL-2, IL-6, and TNF-alpha) and increased the anti-inflammatory cytokine (TGF-beta1), compared to positive control diclofenac (DFC). Urolithin-C effectively abrogated the NF-κB p65 phosphorylation and its translocation to the nucleus as well. Most importantly, Urolithin-C efficiently suppressed the expression of pro-inflammatory genes and elevated the expression of anti-inflammatory gene.

CONCLUSION

Urolithin-C exhibited anti-inflammatory properties by regulating the expression of pro-inflammatory inducible enzyme, cytokines and the translocation of NF-κB p65 to the nucleus.

摘要

目的

在本研究中，对化学合成的尿石素C的抗炎活性进行了评估。

方法

采用化学方法合成尿石素C（3,8,9-三羟基-6H-苯并[c]色烯-6-酮），并使用各种技术对其进行表征。通过膜稳定、蛋白质变性和蛋白酶抑制试验研究合成的尿石素C的抗炎效果。此外，采用MTT（3-[4,5-二甲基噻唑-2-基]-2,5-二苯基溴化四氮唑）试验评估尿石素C的细胞毒性作用。使用脂多糖（LPS）诱导的RAW 264.7（小鼠巨噬细胞）细胞进一步检测尿石素C的抗炎特性。此外，通过酶联免疫吸附测定（ELISA）定量促炎/抗炎细胞因子来研究尿石素C的抗炎特性。使用共聚焦激光扫描显微镜（CLSM）研究尿石素C对核因子κB（NF-κB）易位的作用机制。同时使用逆转录定量聚合酶链反应（RT-qPCR）分析基因表达模式。

结果

与阳性对照阿司匹林相比，尿石素C通过防止溶酶体降解、蛋白质变性和抑制蛋白酶表现出强大的抗炎作用。此外，在较高剂量（200μg/mL）时，发现尿石素C对小鼠巨噬细胞有毒性；然而，在较低浓度（25μg/mL）时，它对上述细胞没有毒性。因此，使用25μg/mL的尿石素C来评估抗炎活性。有趣的是，与阳性对照双氯芬酸（DFC）相比，尿石素C有效降低了促炎诱导酶（Cox-2）、细胞因子（IL-2、IL-6和肿瘤坏死因子-α）的表达，并增加了抗炎细胞因子（转化生长因子-β1）的表达。尿石素C还有效消除了NF-κB p65的磷酸化及其向细胞核的易位。最重要的是，尿石素C有效抑制了促炎基因的表达并提高了抗炎基因的表达。