Publications

@article{InKim2023,

title = {FSLLRY-NH2, a protease-activated receptor 2 (PAR2) antagonist, activates mas-related G protein-coupled receptor C11 (MrgprC11) to induce scratching behaviors in mice},

author = {Hye In Kim and Gi Baek Lee and Da Eun Song and Babina Sanjel and Wook-Joo Lee and Won-Sik Shim},

doi = {10.1016/j.lfs.2023.121786},

issn = {0024-3205},

year  = {2023},

date = {2023-07-00},

urldate = {2023-07-00},

journal = {Life Sciences},

volume = {325},

publisher = {Elsevier BV},

abstract = {Aims: Protease-activated receptor 2 (PAR2), a type of G protein-coupled receptor (GPCR), plays a significant role in pathophysiological conditions such as inflammation. A synthetic peptide SLIGRL-NH2 (SLIGRL) can activate PAR2, while FSLLRY-NH2 (FSLLRY) is an antagonist. A previous study showed that SLIGRL activates both PAR2 and mas-related G protein-coupled receptor C11 (MrgprC11), a different type of GPCR expressed in sensory neurons. However, the impact of FSLLRY on MrgprC11 and its human ortholog MRGPRX1 was not verified. Hence, the present study aims to verify the effect of FSLLRY on MrgprC11 and MRGPRX1.



Methods: The calcium imaging technique was applied to determine the effect of FSLLRY in HEK293T cells expressing MrgprC11/MRGPRX1 or dorsal root ganglia (DRG) neurons. Scratching behavior was also investigated in wild-type and PAR2 knockout mice after injecting FSLLRY.



Key findings: It was surprisingly discovered that FSLLRY specifically activates MrgprC11 in a dose-dependent manner, but not other MRGPR subtypes. Furthermore, FSLLRY also moderately activated MRGPRX1. FSLLRY stimulates downstream pathways including Gαq/11, phospholipase C, IP3 receptor, and TRPC ion channels to evoke an increase in the intracellular calcium levels. The molecular docking analysis predicted that FSLLRY interacts with the orthosteric binding pocket of MrgprC11 and MRGPRX1. Finally, FSLLRY activated primary cultures of mouse sensory neurons, and induced scratching behaviors in mice.



Significance: The present study has revealed that FSLLRY is capable of triggering itch sensation through activation of MrgprC11. This finding highlights the importance of considering the unexpected activation of MRGPRs in future therapeutic approaches aimed at the inhibition of PAR2.},

keywords = {Calcium imaging, FSLLRY, Itch, MrgprC11, MRGPRX1, PAR2, Scratching behavior},

pubstate = {published},

tppubtype = {article}

}

@article{Lee2023,

title = {Rg3-enriched Korean red ginseng alleviates chloroquine-induced itch and dry skin pruritus in an MrgprA3-dependent manner in mice},

author = {Wook-Joo Lee and Won-Sik Shim},

doi = {10.1016/j.imr.2022.100916},

issn = {2213-4220},

year  = {2023},

date = {2023-03-00},

urldate = {2023-03-00},

journal = {Integrative Medicine Research},

volume = {12},

number = {1},

publisher = {Elsevier BV},

abstract = {Background: Previous studies have found that Korean red ginseng extract (KRG) has antipruritic effects, which can be attributed to the presence of Rg3, one of the most potent ginsenosides. Therefore, Rg3-enriched KRG extract (Rg3EKRG) is anticipated to have enhanced antipruritic effects. The present study was conducted to examine the effects of Rg3EKRG in acute chloroquine (CQ)-induced and chronic dry skin pruritus.



Methods: Calcium imaging technique was used in HE293T cells expressing MrgprA3 and TRPA1 ("MrgprA3/TRPA1") and in primary cultures of mouse dorsal root ganglia (DRG) neurons. Mouse scratching behavior tests were performed on dry skin models. To verify the altered expression of itch-related genes, real-time RNA sequencing analysis and PCR were performed on DRG sections obtained from dry skin models.



Results: Rg3EKRG suppressed CQ-induced intracellular calcium changes to a greater degree than KRG. Rg3EKRG dose-dependently inhibited CQ-induced responses in MrgprA3/TRPA1 cells. Rg3EKRG likely targeted MrgprA3 rather than TRPA1 to exert its inhibitory effect. Further, Rg3EKRG strongly inhibited the scratching behavior in mice induced by acute CQ injection. Importantly, DRG neurons obtained from dry skin mice models showed increased mRNA levels of MrgprA3, and treatment with Rg3EKRG alleviated chronic dry skin conditions and suppressed spontaneous scratching behaviors.



Conclusion: The results of the present study imply that Rg3EKRG has a stronger antipruritic effect than KRG, inhibiting both acute CQ-induced and chronic dry skin pruritus in an MrgprA3-dependent manner. Therefore, Rg3EKRG is a potential antipruritic agent that can suppress acute and chronic itching at the peripheral sensory neuronal level.},

keywords = {Dry skin, Itch, Korean Red Ginseng, MrgprA3, RNA-seq, Scratching behavior, TRPA1},

pubstate = {published},

tppubtype = {article}

}

@article{Sanjel2019,

title = {BAM8-22 and its receptor MRGPRX1 may attribute to cholestatic pruritus},

author = {Babina Sanjel and Han-Joo Maeng and Won-Sik Shim},

doi = {10.1038/s41598-019-47267-5},

issn = {2045-2322},

year  = {2019},

date = {2019-12-00},

urldate = {2019-12-00},

journal = {Sci Rep},

volume = {9},

number = {1},

publisher = {Springer Science and Business Media LLC},

abstract = {Pruritus is an unexpected symptom observed in cholestasis and its mechanism is still unclear. Here, we show that bovine adrenal medulla (BAM) 8–22, an endogenous itch-inducing peptide, could be involved in cholestatic pruritus. It was found that bile duct ligation (BDL) mice, an obstructive cholestasis model, showed increased spontaneous scratching behaviour. Importantly, the mRNA level of proenkephalin, a precursor polypeptide of BAM8-22, was significantly increased in the skin of BDL mice. Furthermore, the mRNA level of Mrgprx1, which encodes a receptor for BAM8-22, was significantly increased in the dorsal root ganglia (DRG) of BDL mice. This was further confirmed by elevation of intracellular calcium levels upon BAM8-22 treatment in primarily-cultured DRG neurons. In addition, BDL mice showed augmented scratching behaviour by BAM8-22, indicating enhanced activity of MRGPRX1. Moreover, the skin homogenate of BDL mice induced elevation of intracellular calcium levels through MRGPRX1. Finally, among the various bile acids, chenodeoxycholic acid significantly increased proenkephalin transcription in a human keratinocyte cell line (HaCaT). In conclusion, cholestatic pruritus could be attributed in part to enhanced action of both BAM8-22 in the skin and its receptor MRGPRX1 in sensory neurons.},

keywords = {Animal model, Calcium imaging, Cholestasis, Dorsal root ganglia, MrgprC11, MRGPRX1, Scratching behavior},

pubstate = {published},

tppubtype = {article}

}

@article{Kim2019,

title = {Extracts of the leaves of Pyrus ussuriensis Maxim. Alleviate itch sensation via TSLP-dependent manner in mouse models of atopic dermatitis},

author = {Bo Hyun Kim and Wook-Joo Lee and Babina Sanjel and Kyohee Cho and Youn Kyoung Son and Hye Yoon Park and Sun Yeou Kim and Won-Sik Shim},

doi = {10.1016/j.physbeh.2019.112624},

issn = {0031-9384},

year  = {2019},

date = {2019-10-00},

urldate = {2019-10-00},

journal = {Physiology & Behavior},

volume = {210},

publisher = {Elsevier BV},

abstract = {Pyrus ussuriensis Maxim. commonly known as “Sandolbae” in Korean is a pear tree widely distributed across East Asia. Recent studies indicate that P. ussuriensis Maxim. leaves (PUL) have antipruritic effects. This study aimed to determine the effects of PUL extract and its fractions in decreasing the itch sensation and skin lesions in two distinct animal models of atopic dermatitis (AD) induced by dinitrofluorobenzene (DNFB) or house dust mite(HDM). Our results showed that the total ethanol extract of PUL decreased the scratching behavior in mice withDNFB- and HDM-induced AD. Moreover, the ethyl acetate fraction of PUL significantly improved the overallcondition of the mice with AD induced by HDM. Further, we used HEK293T cells that express receptors and ion channels for thymic stromal lymphopoietin (TSLP), a potent pruritogen for AD, to determine the mechanismsunderlying the antipruritic effects of PUL extract/fractions. Specific subfractions of the PUL strongly inhibited the increase in calcium levels induced by TSLP. In addition, the specific subfraction of PUL inhibited the TSLP-induced increase in calcium levels in cultured mouse dorsal root ganglia neurons. Thus, our results showed thatthe PUL extract could be effective for alleviating pruritus, and the antipruritic effects were exerted probably viathe inhibition of the TSLP pathway in peripheral sensory neurons governing the itch sensation in AD.},

keywords = {Animal model, Atopic dermatitis, Calcium imaging, Itch, Scratching behavior, TSLP},

pubstate = {published},

tppubtype = {article}

}

@article{Lee2018,

title = {Korean Red Ginseng extract and ginsenoside Rg3 have anti-pruritic effects on chloroquine-induced itch by inhibition of MrgprA3/TRPA1-mediated pathway},

author = {Wook-Joo Lee and Young-Sik Kim and Won-Sik Shim},

doi = {10.1016/j.jgr.2017.05.004},

issn = {1226-8453},

year  = {2018},

date = {2018-10-00},

urldate = {2018-10-00},

journal = {Journal of Ginseng Research},

volume = {42},

number = {4},

pages = {470--475},

publisher = {Elsevier BV},

abstract = {Background: It was previously found that Korean Red Ginseng water extract (KRGE) inhibits the histamine-induced itch signaling pathway in peripheral sensory neurons. Thus, in the present study, we investigated whether KRGE inhibited another distinctive itch pathway induced by chloroquine (CQ); a representative histamine-independent pathway mediated by MrgprA3 and TRPA1. Methods: Intracellular calcium changes were measured by the calcium imaging technique in theHEK293T cells transfected with both MrgprA3 and TRPA1 (“MrgprA3/TRPA1”), and in primary culture ofmouse dorsal root ganglia (DRGs). Mouse scratching behavior tests were performed to verify proposed antipruritic effects of KRGE and ginsenoside Rg3.Results: CQ-induced Ca2þ influx was strongly inhibited by KRGE (10 mg/mL) in MrgprA3/TRPA1, and notablyginsenoside Rg3 dose-dependently suppressed CQ-induced Ca2þ influx in MrgprA3/TRPA1. Moreover, bothKRGE (10 mg/mL) and Rg3 (100 mM) suppressed CQ-induced Ca2þ influx in primary culture of mouse DRGs,indicating that the inhibitory effect of KRGE was functional in peripheral sensory neurons. In vivo tests revealed that not only KRGE (100 mg) suppressed CQ-induced scratching in mice [bouts of scratching:274.0  51.47 (control) vs. 104.7  17.39 (KRGE)], but also Rg3 (1.5 mg) oral administration significantly reduced CQ-induced scratching as well [bouts of scratching: 216.8  33.73 (control) vs.115.7  20.94 (Rg3)]. Conclusion: The present study verified that KRGE and Rg3 have a strong antipruritic effect against CQ-induced itch. Thus, KRGE is as a promising antipruritic agent that blocks both histamine-dependent and -independent itch at peripheral sensory neuronal levels.},

keywords = {Calcium imaging, Chloroquine, Korean Red Ginseng, MrgprA3, Scratching behavior, TRPA1},

pubstate = {published},

tppubtype = {article}

}

@article{Ji2018,

title = {Different perception levels of histamine-induced itch sensation in young adult mice},

author = {Yeounjung Ji and Yongwoo Jang and Wook Joo Lee and Young Duk Yang and Won-Sik Shim},

doi = {10.1016/j.physbeh.2018.02.015},

issn = {0031-9384},

year  = {2018},

date = {2018-05-00},

urldate = {2018-05-00},

journal = {Physiology & Behavior},

volume = {188},

pages = {188--193},

publisher = {Elsevier BV},

abstract = {Itch is an unpleasant sensation that evokes behavioral responses such as scratching the skin. Interestingly, it isconceived that the perception of itch sensation is influenced by age. Indeed, accumulating evidence supports theidea that even children or younger adults show distinctive itch sensation depending on age. This evidence implies the presence of a mechanism that regulates the perception of itch sensation in an age-dependent fashion. Therefore, the purpose of the present study was to investigate a putative mechanism for the age-dependent perception of itch sensation by comparing histamine-induced scratching behaviors in 45-day old (D45) and 75-day old male “young adult” mice. The results indicated that, following histamine administration, the D75 mice spent a longer time scratching than D45 mice. However, the intensity of the calcium influx induced by histamine in primary culture of dorsal root ganglia (DRG) neurons was not different between D45 and D75 mice. Moreover, no apparent difference was observed in mRNA levels of a characteristic His-related receptor and ion channel. Incontrast, the mRNA levels of Toll-Like Receptor 4 (TLR4) were increased approximately by two-fold in D75 DRG compared with D45 DRG. Additionally, D75-derived DRG neurons exhibited enhanced intracellular calcium increase by lipopolysaccharide (LPS, a TLR4 agonist) than those of D45 mice. Furthermore, intensities of calciuminflux induced by histamine were significantly potentiated when co-treated with LPS in D75 DRG neurons, butnot in those of D45 mice. Thus, it appears that D75 mice showed enhanced histamine-induced scratching behaviors not by increased expression levels of histamine-related genes, but probably due to augmented TLR4},

keywords = {Dorsal root ganglia, H1R, Histamine, Itch, Scratching behavior, TLR4, TRPV1},

pubstate = {published},

tppubtype = {article}

}