Oriental medicine uses many herbs with biological activity. Among these, some have anti-inflammatory activities, but their action mechanisms have yet to be fully elucidated. Therefore, in this study, we harvested mouse bone marrow cells (BMs) and treated them with lipopolysaccharide (LPS), after which they were treated with the extracts of four medicinal herbs. The metabolic activity, cell death ratio, and tumor necrosis factor (TNF)-alpha production of the BMs was then evaluated. Among the four medicinal herbs, Caesalpinia sappan (CS) significantly decreased the metabolic activity, mitochondrial membrane potential of BMs, and production of TNF-alpha, especially in the presence of LPS. Moreover, CS increased the cell death of BMs stained with propidium iodide. Taken together, these findings indicate that CS inhibited the biological activities of LPS-treated BMs in multiple assays, confirming that CS has anti-inflammatory activity.

Doxorubicin has been used to treating cancers, including breast cancer, bladder cancer, and acute lymphocytic leukemia, however, few studies have investigated its anti-inflammatory activity. In this study, we used mouse spleen cells treated with lipopolysaccharide (LPS), a representative inflammatory agent to investigate the effects of doxorubicin. Specially, we investigated the effects of doxorubicin on metabolic activity, cell size, cell death, and cytokine production of LPS-treated spleen cells. Doxorubicin significantly decreased the metabolic activity, even when applied at relatively low concentrations (1.6-8 ng/mL). To investigate the potential mechanism, we measured the mitochondrial membrane potential (MMP) of the LPS-treated spleen cells using Rhodamine 123. Doxorubicin decreased MMP and cell size, and induced cell death. Furthermore, doxorubicin suppressed the production of tumor necrosis factor (TNF)-alpha, a representative cytokine, in LPS-treated spleen cells. Taken together, doxorubicin decreased metabolic activity and the production of inflammatory cytokines, while increasing the death of LPS-induced hyperactivated spleen cells. This results will enable broader application of doxorubicin, as an anti-inflammatory agent, in clinical and research fields.

Transglutaminase2 (TGM2) is a multi-functional calcium dependent enzyme that affects angiogenesis, apoptosis, differentiation, attachment, and changes in the extracellular matrix. However, its function in periodontal tissue has not yet been studied. The aim of this study was to investigate the association of the TGM2 expression and the modulation of inflammatory mediators in inflamed periodontal ligament (PDL) cells induced by pro-inflammatory cytokines such as Interleukin-1β and the Tumor necrosis factor-α. The expression of TGM2 was increased in the inflamed periodontal tissue and PDL cells. Over-expressed TGM2 in the PDL cells increased expression of MMP1, MMP3, IL-6, CXCL8, and PTGS2. Conversely, inhibition of TGM2 activity using LDN27219, a TGM2 inhibitor, resulted in decreased expression of MMP1, MMP3, IL-6, and CXCL8. The mRNA expression was confirmed by RT-PCR and quantified by qRT-PCR. Protein levels were also confirmed by immunofluoroscence staining. These results suggest that TGM2 plays an important role in the regulation of inflammatory mediators which exacerbate tissue damage in inflamed periodontal tissue.

Bropirimine, a class of antineoplastic agents, is known as one of the potent immunomodulators and is currently under clinical development for the treatment of cancer. However, the effect of bropirimine on the cow remains unknown as a therapeutics agent. In this experiment, the effect of bropirimine in the peripheral blood mononuclear cells (PBMCs) stimulated by lipopolysaccharide (LPS) or concanavalin-A (Con-A) was examined. Jugular venous blood was collected from Korean Hanwoo calves and PBMCs were isolated. It was used to study the effect of bropirimine upon stimulation with LPS or Con-A for 72 hours. The expression pro-inflammatory cytokines like Tumor Necrosis Factor α (TNF-α) and Interferon γ (IFN-γ) were confirmed. Bropirimine significantly inhibited LPS- or Con-A-induced TNF-α and Con-A-induced IFN-γ in dose-dependent manner. Furthermore, Bropirimine inhibited TNF-α and Con-A mRNA expression at the transcription level. These results clearly indicated that bropirimine inhibited LPS or Con-A stimulated up-regulation of proinflammatory cytokines in a dose-dependent manner without conspicuous cytotoxicity. The bropirimine has potential to protect cow from LPS or Con-A induced endotoxin shock, possibly through inhibition of the production of proinflammatory cytokines. It suggesting that bropirimine may be a novel therapeutic agent for the prevention of inflammatory diseases. This result revealed specific features of the immune responses depending on the bropirimine compound and would help to knowledge of bovine immunity.

We have previously shown that the specific phosphatidylinositol 3-kinase inhibitor LY294002 (LY29), and its inactive analog LY303511 (LY30), inhibit a monocyte chemoattractant protein-1 (MCP-1) expression in human umbilical vein endothelial cells; these results suggest the potential of LY30 as an anti-inflammatory drug. In this study, we determined the effects of LY30 on the production of various inflammatory cytokines in human macrophagic THP-1 cells which were stimulated with lipopolysaccharide (LPS). LY30 selectively suppressed the mRNA expression of IL-12 p40, TNF-α, and MCP-1 without affecting the expression of IL-1α, IL-6, and IL-8. Inhibition of the production of IL-12 and TNF-α by LY30 was also demonstrated using ELISA assays. In order to elucidate the mechanisms of the action of LY30, we examined the role played by the mitogen-activated protein kinases and the key transcription factors, AP-1 and NF-κB in LPS-stimulated THP-1 cells. The results revealed that LY30 inhibited LPS-induced activation of ERK, but not p38 or JNK. Furthermore, the AP-1 DNA binding activity was suppressed by LY30 based upon the dosage, whereas NF-κB DNA binding was not affected. These results suggest that LY30 selectively inhibits cytokine production in the LPS-stimulated macrophagic THP-1 cells by downregulating the activation of ERK and AP-1.

Background: Periodontitis is an inflammatory disease characterized by the breakdown of tooth-supporting tissues, leading to tooth loss. Aggregatibacter actinomycetemcomitans are major etiologic bacterium causing aggressive periodontitis. Ursodeoxycholic acid (UDCA), a hydrophilic gall bladder acid, has been used as an effective drug for various diseases related to immunity. The aim of this study was to investigate the effect of UDCA on the inflammatory response induced by A. actinomycetemcomitans. Methods: A human acute monocytic leukemia cell line (THP-1) was differentiated to macrophage- like cells by treatment with phorbol 12-mystristate 13-acetate (PMA) and used for all experiments. The cytotoxic effect of UDCA was examined by MTT assay. THP-1 cells were pretreated with UDCA for 30 min before A. actinomycetemcomitans infection and the culture supernatant was analyzed for various cytokine production by ELISA. The effect of UDCA on bacterial growth was examined by measuring optical densities using a spectrophotometer. Results: UDCA showed no cytotoxic effect on THP-1 cells, up to 80 μM Ed highlight: Please confirm technical meaning. UDCA pretreatment inhibited the A. actinomycetemcomitansinduced IL-1β, TNF-⍺, and IL-17A secretion in a dosedependent manner. UDCA also inhibited IL-21 production at 60 μM. The production of IL-12 and IL-4 was not influenced by A. actinomycetemcomitans infection. Conclusion: These findings indicate that UDCA inhibits the production of inflammatory cytokines involved in innate and Th17 immune responses in A. actinomycetemcomitansinfected THP-1- derived macrophages, which suggests its possible use for the control of aggressive periodontitis.

Many kinds of medicinal herbs have been used to treat inflammation in Oriental medicine. However, there few studies have investigated the anti-inflammatory activity of medicinal herbs. In this study, we used mouse bone marrow cells (BMs) treated with lipopolysaccharide (LPS), a simulator of osteomyelitis, to screen medicinal herbs having anti-inflammatory activity. Specifically, we investigated the activity of an extract of Rhus chinensis (RC) using metabolic activity and cytokine production of the BMs treated with LPS and RC. The metabolic activity of BMs was measured using Cell Counting Kit-8® solution. RC decreased the metabolic activity of LPS-treated BMs. A viability assay using trypan blue solution demonstrated that RC marginally decreased the viability of LPS-treated BMs. Flow cytometry analysis revealed that RC decreased the mitochondrial membrane potential of BMs, regardless of LPS treatment. To investigate the anti-inflammatory activity of RC, we measured the production of tumor necrosis factor (TNF)-alpha and interleukin (IL)-10 in BMs. LPS increased the production of both cytokines in BMs. Interestingly, RC induced a greater increase in IL-10 than TNF-alpha in LPS-treated BMs. Taken together, RC decreased metabolic activity and modulated the production of inflammation-related cytokines in LPS-treated BMs. These findings suggest that RC can be used as a medicinal herb with anti-inflammatory activity.

The objective of this study was to investigate the effects of a lettuce (Lactuca sativa L.) extract on the inflammation of human umbilical vein endothelial cell (HUVEC) and blood lipid improvement in hypercholesterolemic mice fed a high cholesterol diet. The lettuce extract (100% ethanol extract) inhibited the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in HUVEC treated with tumor necrosis factor-α (TNF-α). The lettuce extract suppressed the adhesion of THP-1 to TNF-α-treated HUVEC. The lettuce extract decreased the TNF-α-stimulated production of proinflammatory cytokine interleukin-6, interleukin-8 and chemokine monocyte chemotactic protein 1. In hypercholesterolemic mice, the lettuce extract reduced serum total cholesterol, triglyceride, and low-density lipoprotein-cholesterol level, while the lettuce extract elevated high-density lipoprotein-cholesterol level, resulting in the decrease of atherogenic index and cardiac risk factor level. These results suggested that lettuce extract can be an useful resource to show an anti-inflammatory effect and improve lipid metabolism

Bone fractures are most often seen in racetrack horses because of the high level of intensity in racing. These issues are the main cause of decreased performance in racehorses. Mesenchymal stem cells (MSCs) have been explored to improve intra-articular therapy in racehorses. MSCs are essential for the repair and regeneration of damaged tissues. In this study, the effect of intra-articular injection of MSCs in racehorses was investigated. Before accessing the MSC therapy, synovial fluids were obtained from the fracture site of racehorses, and adipose tissue was collected for MSC isolation. Using the MSC specific marker, adipose tissue-derived MSCs were identified. The racehorses received intra-articular injection of autologous MSCs (or allogeneic) (3 × 107 cells/3 mL). After 1 or 2 weeks, synovial fluids were collected from racehorses. To test the effect of MSC injection using ELISA, we analyzed inflammatory factors from the untreated samples compared to MSC-treated samples of racehorses. The level of pro-inflammatory factors (interleukin-1β and prostaglandin E2) was significantly decreased in synovial fluids of MSC-injected racehorses, compared to before accessing the MSC therapy, whereas, the level of anti-inflammatory factor (interleukin-10) was higher than prior to accessing the MSC therapy. Further studies are needed to investigate the anti-inflammatory mechanism of MSC in racehorses.

제주산 식용 고사리를 열수와 70% 에탄올로 추출한 후, 70% 에탄올 추출물을 순차적으로 헥산, 에틸아세테이트, 부탄올, 증류수로 분획하여 폴리페놀 함량과 항산화 활성 및 NO 생성 저해효과를 측정하였다. 총페놀 함량은 열수와 70% 에탄올 추출물이 각각 50.0과 43.4mg GAE/g이었고, 에틸아세테이트와 부탄올 분획물에서 각각 104.3과 58.7mg GAE/g로 높았다. DPPH 라디칼 소거활성(IC50)은 에틸아세테이트 분획물이 100.6μg/mL로 가장 높았고, 그 다음으로 부탄올 분획물(182.6), 70% 에탄올 추출물(231.5) 순이었다. ABTs 라디칼 소거활성(IC50)은 열수와 70% 에탄올 추출물이 각각 102.4와 119.4μg/mL이었고, 분획물에서는 에틸아세테이트가 70.7μg/mL으로 가장 높았으며 그 다음으로 부탄올(141.7)이었다. 활성산소 흡수능력(ORAC)은 열수와 70% 에탄올 추출물이 각각 267.8과 205.5mg TE/g이었고, 에틸아세테이트 분획물이 836.0mg TE/g로 가장 높았으며, 부탄올 분획물은 337.9mg TE/g이었다. NO 생성 저해활성(IC50)은 에틸아세테이트 분획물이 24.9μg/mL로 가장 높았으며, 그 다음으로 헥산과 부탄올 분획물이 34.9와 118.0μg/mL이었다. 결론적으로 제주산 고사리의 에틸아세테이트 분획물은 총페놀 함량은 물론 항산화 활성과 항염 활성이 가장 높아 기능성 소재로 활용될 수 있을 것으로 추정되었다.

The activation of glial cells in the spinal cord has been contribute to the initiation and maintenance of pain facilitation induced by peripheral inflammation and nerve injury. The present study investigated effects of botulinum toxin type A (BoNT-A), injected subcutaneously or intracisternally, on the expression of microglia and astrocytes in rats. Complete Freund’s Adjuvant (CFA)-induced inflammation was employed as an orofacial chronic inflammatory pain model. A subcutaneous injection of 40 μL CFA into the vibrissa pad was performed under 3% isoflurane anesthesia in SD rats. Immunohistochemical analysis for changes in Iba1 (a microglia marker) and GFAP (an astrocyte marker), were performed 5 days after CFA injection. Subcutaneous injection of CFA produced increases in Iba1 and GFAP expression, in the ipsilateral superficial lamia I and II in the medullary dorsal horn of rats. Subcutaneous treatment with BoNT-A attenuated the up-regulation of Iba1 and GFAP expressions induced by CFA injection. Moreover, intracisternal injection of BoNT-A also attenuated the up-regulated Iba1 and GFAP expressions. These results suggest that the anti-nociceptive action of BoNT-A is mediated by modulation activation of glial cells, including microglia and astrocyte.

최근 알로에 베라 섭취에 의한 독성 간염이 보고되고 있으나, 아직까지 알로에 베라의 간에 대한 염증 효과가 명확히 밝혀지지 않았다. 본 연구는 알로에 베라 에탄올 추출물이 간세포의 염증 발현에 미치는 영향과 그 기전을 밝히기 위해 수행되었다. 0.001~100 μg/mL의 알로에 베라 추출물을 HepG2세포에 처리하여 MTT assay를 시행한 결과, 모든 농도에서 세포사멸이 유도되지 않았다. 그러나 모든 농도에서 알로에 베라 추출물은 염증성 사이토카인인 IL-8의 분비를 15.7~25.8%까지 유의적으로 증가시켰다(p<0.05). 또 다른 사이토카인인 M-CSF도 알로에 베라 추출물에 의해 36.3~61.5%까지 유의적으로 분비가 증가되었다(p<0.05). 본 연구 결과는 알로에 베라 추출물이 IL-8과 M-CSF와 같은 염증성 사이토카인 분비기전에 의해 간에 염증을 유발할 수 있음을 보여준다. 또한 알로에 추출물에 의해 유발될 수 있는 간염기전을 제시함으로써, 향후 수행될 추가 실험을 위한 기초 자료로 그 가치가 높을 것으로 사료된다.

Tyrophagus putrescentiae (Tp) as a storage mite inhabitats in stored grains, hay, and straw at agricultural areas. T. putrescentiae stimulates an immune response and triggers inflammatory cytokines release, and thus it is a source of allergen that sensitize and induce allergic reactions. Also, T. putrescentiae has been reported to cause asthma and atopic disease by cross-reactivity with Dermatophagoides pteronyssinus (Dp). The study on T. putrescentiae in human monocytic THP-1 cells is not enough to understand cytokine expression and pathological mechanisms. The aim of this study is to investigate the effect of T. putrescentiae extract (TpE) on production of inflammatory cytokines and expression of mRNA level in THP-1 cells. THP-1 cells are treated with TpE and supernatants were analyzed for the production of cytokines using enzyme-linked immunosorbent assay (ELISA). mRNA level in the culture cells was measured by a reverse transcriptase-polymerase chain reaction (RT-PCR). As a result of this study, TpE significantly induced secretion of interleukin-6, interleukin-8, and monocyte chemotactic protein-1 (MCP-1) in THP-1 cells in time- and dose-dependent manner. These results suggest that TpE may play a role in contributing to inflammatory disease through stimulation of immune cell. Further research of T. putrescentiae is needed to understand the elucidation of the pathogenic mechanism.