# Application of 18β-glycyrrhetinic acid Fluorescent probes in cell imaging

**Authors:** Yifan Zhao, Hongxia Sun, Zhiwei Yan, Youde Wang, Shuai Li, Yachun Guo, Guangxin Miao, Tienan Wang, Liying Zhang, Chengjun Song

PMC · DOI: 10.1080/14756366.2026.2631869 · 2026-03-09

## TL;DR

Researchers developed fluorescent probes to study how 18β-GA reduces inflammation in macrophages, finding that it likely acts on cytoplasmic proteins.

## Contribution

Designed and tested fluorescent probes to identify the subcellular targets of 18β-GA in anti-inflammatory processes.

## Key findings

- Modifying C-3 hydroxyl and C-30 carboxyl groups improved 18β-GA's anti-inflammatory activity.
- Two fluorescent probes (Ia and IIc) showed similar anti-inflammatory effects to 18β-GA on key inflammatory factors.
- Fluorescence signals from the probes were localized in the cytoplasm, suggesting cytoplasmic protein targets.

## Abstract

Fluorescently labelled small molecule probes (fluorescent probes) play an important role in cell imaging and are often used in combination with light-affinity probes to determine the subcellular localisation of target proteins. To investigate the target proteins of 18β-glycyrrhetinic acid (18β-GA), which regulates the macrophage inflammatory response, we designed and synthesised three types of fluorescent probes. We analysed its structure-activity relationship by evaluating the biological activity and screening for fluorescent probes with high activity. Our results showed that modifying C-3 hydroxyl and C-30 carboxyl groups enhanced the anti-inflammatory activity of 18β-GA, and found that two preferred probes had similar effects on LPS-induced, inflammation-related factor release (IL-1β, TNF-α, IL-6, HDAC8, P-STAT3, and SOCS3) to those of 18β-GA. Fluorescence signals of the preferred probes Ia and IIc were observed in the cytoplasm. The above results indicated that the anti-inflammatory site of 18β-GA may be located in proteins in the cytoplasm, which would provide useful information for research on the anti-inflammatory targets of 18β-GA.

## Linked entities

- **Proteins:** HDAC8 (histone deacetylase 8), SOCS3 (suppressor of cytokine signaling 3)
- **Chemicals:** 18β-glycyrrhetinic acid (PubChem CID 10114), IL-6 (PubChem CID 165368475)

## Full-text entities

- **Genes:** App (amyloid beta precursor protein) [NCBI Gene 11820] {aka Abeta, Abpp, Adap, Ag, Cvap, E030013M08Rik}, Hdac8 (histone deacetylase 8) [NCBI Gene 70315] {aka 2610007D20Rik}, Pik3r1 (phosphoinositide-3-kinase regulatory subunit 1) [NCBI Gene 18708] {aka PI3K, p50alpha, p55alpha, p85alpha}, Il6 (interleukin 6) [NCBI Gene 16193] {aka Il-6}, HDAC8 (histone deacetylase 8) [NCBI Gene 55869] {aka CDA07, CDLS5, HD8, HDACL1, KDAC8, MRXS6}, STAT3 (signal transducer and activator of transcription 3) [NCBI Gene 6774] {aka ADMIO, ADMIO1, APRF, HIES}, Nfkb1 (nuclear factor of kappa light polypeptide gene enhancer in B cells 1, p105) [NCBI Gene 18033] {aka NF-KB1, NF-kappaB, NF-kappaB1, p105, p50, p50/p105}, TNF (tumor necrosis factor) [NCBI Gene 7124] {aka DIF, IMD127, TNF-alpha, TNFA, TNFSF2, TNLG1F}, Il1b (interleukin 1 beta) [NCBI Gene 16176] {aka IL-1beta, Il-1b}, Socs3 (suppressor of cytokine signaling 3) [NCBI Gene 12702] {aka Cis3, Cish3, EF-10, Ef10, SSI-3, Ssi3}, IL1B (interleukin 1 beta) [NCBI Gene 3553] {aka IL-1, IL1-BETA, IL1F2, IL1beta}, Stat3 (signal transducer and activator of transcription 3) [NCBI Gene 20848] {aka 1110034C02Rik, Aprf}, Akt1 (Akt serine/threonine kinase 1) [NCBI Gene 11651] {aka Akt, LTR-akt, PKB, PKB/Akt, PKBalpha, Rac}, Gapdh (glyceraldehyde-3-phosphate dehydrogenase) [NCBI Gene 14433] {aka Gapd}, SOCS3 (suppressor of cytokine signaling 3) [NCBI Gene 9021] {aka ATOD4, CIS3, Cish3, SOCS-3, SSI-3, SSI3}, Tnf (tumor necrosis factor) [NCBI Gene 21926] {aka DIF, TNF-a, TNF-alpha, TNFSF2, TNFalpha, Tnfa}, Cort (cortistatin) [NCBI Gene 12854] {aka CST, PCST}, IL6 (interleukin 6) [NCBI Gene 3569] {aka BSF-2, BSF2, CDF, HGF, HSF, IFN-beta-2}, Nos2 (nitric oxide synthase 2, inducible) [NCBI Gene 18126] {aka MAC-NOS, NOS-II, Nos-2, Nos2a, i-NOS, iNOS}
- **Diseases:** Pneumonia (MESH:D011014), Chronic inflammation (MESH:D007249), respiratory disease (MESH:D012140), vascular diseases (MESH:D014652), inflammatory drugs (MESH:D000081015), allergies (MESH:D004342), infectious (MESH:D003141), infection (MESH:D007239), macrophage (MESH:D055501)
- **Chemicals:** amide (MESH:D000577), water (MESH:D014867), 18beta-GA (MESH:C119129), brine (MESH:C017082), 18-crown-6 (MESH:C015762), sodium ascorbate (MESH:D001205), SDS (MESH:D012967), NaOH (MESH:D012972), 13C (MESH:C000615229), 4-benzoylbenzoic acid (MESH:C058818), glycyrrhizin (MESH:D019695), silica gel (MESH:D058428), 3H (MESH:D014316), DNS-Cl (MESH:C029038), ABP (MESH:C072526), FITC (MESH:D016650), 3-azidopropan-1-amine (MESH:C000625736), polyethylene glycol (MESH:D011092), N2 (MESH:D009584), azide (MESH:D001386), 3-Bromopropyne (MESH:C458239), streptomycin (MESH:D013307), triazole (MESH:D014230), CO2 (MESH:D002245), LPS (MESH:D008070), paraformaldehyde (MESH:C003043), Cu (I) (MESH:C073870), 2-bromoethanol (MESH:C025475), DIPEA (MESH:C027070), THF (MESH:C018674), EDCI (MESH:D005022), PBS (MESH:D007854), GA (MESH:D005708), H (MESH:D006859), alkyne (MESH:D000480), 2H (MESH:D003903), 2-(2-bromoethoxy)ethanol (-), NaHCO3 (MESH:D017693), K2CO3 (MESH:C037593), penicillin (MESH:D010406), Na (MESH:D012964), potassium tert-butoxide (MESH:C077664), Na2SO4 (MESH:C012036), TFA (MESH:D014269), TMS (MESH:D013932), oil (MESH:D009821)
- **Species:** Mus musculus (house mouse, species) [taxon 10090], Glycyrrhiza (licorice, genus) [taxon 46347], Homo sapiens (human, species) [taxon 9606]
- **Mutations:** P0013C, G6203A
- **Cell lines:** RAW 264.7 — Mus musculus (Mouse), Mouse leukemia, Cancer cell line (CVCL_0493)

## Figures

11 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12973793/full.md

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Source: https://tomesphere.com/paper/PMC12973793