Jong-Soon Lim

Immune-suppressive activity of punicalagin via inhibition of NFAT activation.

Since T cell activation is central to the development of autoimmune diseases, we screened a natural product library comprising 1400 samples of medicinal herbal extracts, to identify compounds that suppress T cell activity. Punicalagin (PCG) isolated from the fruit of Punica granatum was identified as a potent immune suppressant, based on its inhibitory action on the activation of the nuclear factor of activated T cells (NFAT). PCG downregulated the mRNA and soluble protein expression of interleukin-2 from anti-CD3/anti-CD28-stimulated murine splenic CD4+ T cells and suppressed mixed leukocytes reaction (MLR) without exhibiting cytotoxicity to the cells. In vivo, the PCG treatment inhibited phorbol 12-myristate 13-acetate (PMA)-induced chronic ear edema in mice and decreased CD3+ T cell infiltration of the inflamed tissue. These results suggest that PCG could be a potential candidate for the therapeutics of various immune pathologies.

S100A8 and S100A9 are messengers in the crosstalk between epidermis and dermis modulating a psoriatic milieu in human skin

S100A8 and S100A9 are members of the S100A8 protein family that exist as homodimers and heterodimers in neutrophils, monocytes, and macrophages. Recent studies have shown the pivotal roles of S100A8 and S100A9 in the propagation of inflammation and keratinocyte proliferation in psoriasis. We found significant up-regulation of S100A8 and S100A9 secretion from keratinocytes in psoriatic lesions. To mimic the in vivo secretory conditions of S100A8 and S100A9 from psoriatic epidermal keratinocytes, we used the culture medium (CM) of S100A8 and S100A8/A9 adenovirus-transduced keratinocytes to investigate the functions of S100A8 and S100A9. We detected increased levels of various pro-inflammatory cytokines in the CM, including IL-8 and TNF-α, which are involved in aggravating psoriatic skin lesions, and IL-6 and members of the CXCL family of pro-angiogenic cytokines. The CM increased immune cell migration and increased angiogenesis in human umbilical vein endothelial cells. In conclusion, we found that the upregulated production of S100A8 and S100A9 by psoriatic epidermal keratinocytes activated adjacent keratinocytes to produce several cytokines. Moreover, S100A8 and S100A9 themselves function as pro-angiogenic and chemotactic factors, generating a psoriatic milieu in skin.

Oxidation of polycyclic aromatic hydrocarbons by laccase of Coriolus hirsutus

The oxidation of five polycyclic aromatic hydrocarbons; anthracene, benzo(α)pyrene, fluoranthene, phenanthrene and pyrene was catalyzed by laccase from Coriolus hirsutus in the presence of the redox mediators, 2,2′-azinobis(3-ethylbenzthiazoline-6-sulfonate) (ABTS) and 1-hydroxybenzotriazole (HBT). In the ABTS-mediated system, benzo(α)pyrene was the most rapidly oxidized substrate, with anthracene being the most rapidly oxidized in the HBT-mediated system. There was no clear relationship between the ionization potential and the oxidation of the substrates. ABTS increased the oxidation of benzo(α)pyrene more than HBT but the oxidation of the other PAHs tested were the opposite. The mediators used in conjunction increased the oxidation of benzo(α)pyrene compared to using the mediators alone.

A Global Gene Expression Analysis of the Peripheral Blood Mononuclear Cells Reveals the Gene Expression Signature in Psoriasis

BACKGROUND Psoriasis is a chronic inflammatory skin disease that affects approximately 1~3% of the general population. OBJECTIVE We performed cDNA microarray analysis with using the dendrimer labelling method to investigate the gene expression profile in the peripheral blood mononuclear cells (PBMCs) of psoriatic patients. METHODS The peripheral blood mononuclear cells of 5 patients with psoriasis and 8 control subjects were used in the gene expression analyses of psoriasis. RESULTS We identified 212 differentially expressed genes that showed at least a two-fold induction and/or reduction in psoriatic patients. Among those, 63 genes, including CD44, CD56 and IL7R, were induced, while 139 genes, including the sphingosine kinase 1 and p16-INK genes, were reduced in the psoriatic patients. CONCLUSION We can speculate that these genes may have a role for the pathogenesis of psoriasis via their affecting different cellular functions. Our results suggest a possible mechanism by which activated immune cells migrate from the blood to the skin in psoriatic patients, and we provide novel putative targets for developing drugs to treat psoriasis.

Absence of a human DnaJ protein hTid-1S correlates with aberrant actin cytoskeleton organization in lesional psoriatic skin

Background: HSP27 phosphorylation plays pivotal roles on F-actin polymerization and actin cytoskeleton organization. Results: The loss of hTid-1S expression was observed in lesional human psoriatic skin. Conclusion: The binding of hTid-1S with MK5 inhibits HSP27 phosphorylation and attenuates F-actin polymerization. Significance: The lack of hTid-1S expression correlates with the aberrant actin cytoskeleton organization in psoriatic human skin. The biochemical mechanism by which the human tumorous imaginal disc1S (hTid-1S) interferes with actin cytoskeleton organization in keratinocytes of human skin epidermis was investigated. We found that hTid-1, specifically hTid-1S, interacts with MK5, a p38-regulated/activated protein kinase, and inhibits the protein kinase activity of MK5 that phosphorylates heat shock protein HSP27 in cultured HeLa cells. Thus, hTid-1S expression inhibits the phosphorylation of HSP27 known to play important roles in F-actin polymerization and actin cytoskeleton organization. The interplay between MK5/HSP27 signaling and hTid-1S expression was supported by the inhibition of HSP27 phosphorylation and MK5 activity in HeLa cells in response to hypoxia during which hTid-1S expression was down-regulated. We also found that overexpression of hTid-1S results in the inhibition of HSP27 phosphorylation, F-actin polymerization, and actin cytoskeleton organization in transduced HaCaT keratinocytes. This study further proposes that the loss of hTid-1S expression in the basal layer of skin epidermis correlates with the enhanced HSP27 phosphorylation, keratinocyte hyperproliferation, and excess actin cytoskeleton organization in lesional psoriatic skin.

Expression of Neutrophil Gelatinase-Associated Lipocalin in Calcium-Induced Keratinocyte Differentiation

In a previous search for the differentially expressed genes in keratinocyte differentiation, we identified neutrophil gelatinase-associated lipocalin (NGAL) as a calcium-induced gene. In this study, we further verified the expression of NGAL in cultured keratinocytes as well as in several skin diseases. Reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and ELISA clearly showed that NGAL expression was markedly increased in calcium-induced keratinocyte differentiation in vitro. However, in our previous report, NGAL expression was not detected in normal skin tissue except for hair follicle by in situ hybridization and immunohistochemistry, indicating the difference of cell status between in vitro and in vitro conditions. Interestingly, NGAL expression was highly increased in psoriasis-like inflammatory disorders (lichen planus and pityriasis rubura pilaris) and skin cancers (keratoacanthoma and squamous cell carcinoma), implying that NGAL may be related with the epidermal hyperplasia. Collectively, these results reveal the potential importance of NGAL in the maintenance of skin homeostasis.

Screening and isolation of a natural dopamine D1 receptor antagonist using cell-based assays.

To develop a cell-based assay to screen for human dopamine D(1) receptor agonists or antagonists from medicinal plant extracts, a stable Chinese hamster ovary (CHO) cell line (CHO-D1R) expressing the human dopamine D(1) receptor was established using an expression vector containing a scaffold attachment region (SAR) element. CHO-D1R cells showed specific binding to [(3)H]-SCH23390 with high affinity (K(d)=1.47+/-0.17 nM) and dose-dependent responses for the dopamine-mediated stimulation of cAMP concentrations (EC(50)=20.6+/-1.44 nM). The screening of medicinal plant extracts using cell-based cAMP assays revealed that an extract of Gleditsia sinensis Lam., which is known to be rich in saponin, had strong antagonist activity for the D(1) receptor. From the activity-guided fractionation and chemical structural analysis of the G. sinensis extract, a compound called gleditsioside F was isolated and was identified to have antagonist activity for the D(1) receptor. Gleditsioside F showed very effective D(1) antagonist activity by inhibiting ligand binding to the D(1) receptor as well as by inhibiting dopamine-mediated increases in cAMP concentration.

Single Oral Dose Toxicity Test of ACM(Added Chongmyung-tang) in Sprague-Dawley Rat

AbstractObjectives :This research investigates the single oral dose toxicity of ACM in SD rats.Methods :ACMs were administered to female and male SD rats, as an oral d ose of 5000 ㎎/㎏. Animals were monitored for the mortality and changes in the body weight, clinical signs and gross observation during the 14 days after dosing, upon necropsy. Results :We could not find any mortality. Compared with the control grou p, significant weight change was not observed in the experimental group. First day after administration, compoun d-colored stool was observed in all rats. After the second day of administration, the more common symptoms were not observed . There were no gross abnormalities in all cases. [ED NOTE: highlight: given the context, this is very vague]Conclusions :The results obtained in this study suggest that the approximate lethal dose of ACM in both female and male rats were considered as over 5000 ㎎/㎏. Key Words:ACM, Added Chongmyung-Tang, single oral dose toxicity, SD rats Received : May 25, 2012; Revised : June 07, 2012; Accepted : Ju ne 21, 2012Correspondence : Sang-Ryong Lee, Address : Daejon Univ. College of Oriental Medicine, Yongun-dong, Dong-gu, Daejeon, KoreaTel : 043-229-3726, Fax : 043-253-8757, E-mail : 7575np@dju.kr*This study was supported by a grant of the Traditional Korean Medicine R&D Project, Ministry of Health & Welfare, Republic of Korea. (Task-specific number: B090020)

A Thirteen Week Repeated Oral Dose Toxicity Test and A Four Week Recovery Test of ACM(Added Chongmyung-tang) in Sprague-Dawley Rats

AbstractObjectives :To provide information on the safety of ACM, we carried out a 13-week repeated oral dose toxicity and a 4-week recovery test of ACM in Sprague-Dawley rats.Methods :Female and male rats were treated with ACM with oral doses of 800, 2000, and 5000 ㎎/㎏. The ACM was administered for 13 weeks. Mortality, clinical signs, body weig ht changes, food consumption, ophthalmologic findings, urinalysis, hematological and biochemical parameters, gross findings, organ weights and histological markers were monitored during the study period. Moreover, the rats were monitored for 4 extra weeks to determine recovery time after the study period.Results :We found no mortality and no abnormalities in clinical signs, b ody weight, food consumption, ophthalmologic findings, urinalysis, hematological and biochemical parameters, gross findings, organ weights and histological markers in any of the rats tested.Conclusions :The no-observed adverse effects level (NOAEL) was considered as over 5000 mg/kg for male and female rats.Key Words :ACM, Added Chongmyung-Tang, 13 week repeated oral dose toxicity test, 4 week recovery test, SD rats. Received : August 17, 2012; Revised : September 13, 2012; Accep ted : September 13, 2012Correspondence : In-Chul Jung, Address : Daejon Univ. College o f Oriental Medicine, Yongun-dong, Dong-gu, Daejeon, Korea.Tel : +82-42-470-9129, Fax : +82-42-470-9006, E-mail : npjeong @dju.ac.kr* This study was supported by agrant of the Traditional Korean Medicine R&D Project, Ministry of Health & Welfare, Republic of Korea. (Task-specific number: B090020).

Inhibitory Effects of Soyeum Pharmacopuncture (SPP) on Rheumatoid Arthritis in Collagen II-induced Arthritis (CIA) Mice

Objective : The aim is to examine the effect of Soyeum Pharmacopuncture (SPP) on collagen-induced arthritis (CIA) in DBA/1OlaHsd mice. Methods : To determine the effect of SPP on chronic IFNlammatory joint disease, we induced CIA in DBA/1OlaHsd mice by immunization with bovine type II collagen. Animals were treated with intraperitoneal injection doses of 2 mg/kg of SPP, beginning 3 days before the expected onset of disease symptoms. Inhibitory Effects of SPP were observed by serum levels of TNF-, and IFN-,,, or cell proliferation in the spleen cell culture and histological examination of knee joint. Results : In the CIA Mice, serum levels of TNF-, and IFN-,, production in the spleen cell culture were reduced. At the histopathological examination of knee joint, chondropathy of cartilage in the synovial joint in the SP group was repaired while compared with control group. Conclusion : These results suggest that the SPP may be effective for the prevention and treatment of rheumatoid arthritis disease.