Periodontal disease (PD) is strongly linked to increased risk of oral squamous cell carcinoma (OSCC); however, the specific mechanism through which the development of PD and OSCC is simultaneously promoted remains unclear. This study explored the impact of periodontal pathogens on OSCC progression and the contribution of periodontal pathogen-stimulated OSCC to PD development. The expression of osteoclastogenesis-inducing factors was assessed using quantitative reverse transcription polymerase chain reaction analysis following stimulation of OSCC with lipopolysaccharide (LPS) derived from the periodontal pathogen Porphyromonas gingivalis (Pg), a pathogen commonly responsible for PD. The cell counting kit-8 assay was used to determine the effects of Pg-LPS on OSCC proliferation and drug resistance to cisplatin and 5-fluorouracil. The effects of conditioned medium (CM) derived from Pg-LPS–stimulated OSCC on osteoclastogenesis was evaluated using tartrate-resistant acid phosphatase (TRAP) staining on bone marrow-derived macrophages (BMMs). Pg-LPS administration in SCC-25 and YD-8 OSCC cell lines induced a significant increase in receptor activator of nuclear factor kappa-B ligand mRNA expression; however, it did not affect cell proliferation. Treatment with CM derived from Pg-LPS–stimulated SCC-25 or YD-8 cells markedly enhanced the formation of TRAP-positive multinucleated cells during osteoclast differentiation of BMMs. Altogether, these findings demonstrate that Pg-LPS–stimulated OSCC promoted osteoclastogenesis through a paracrine mechanism.

Brassica oleracea var. italica (broccoli) is a type of cabbage that contains vitamins, minerals, and phytochemicals. Consequently, it is used as a potential nutraceutical source for improving human health by reducing oxidative stress and inflammatory responses. Here, the effects of broccoli sprout extract (BSE) on the inflammatory response were investigated through lipopolysaccharide (LPS)- induced inflammatory mouse models. First, we found that the BSE obviously reduce NO production in RAW 264.7 cells in response to LPS stimulation in in vitro study. Pretreatment with BSE administration improved sperm motility and testicular cell survivability in LPS-induced endotoxemic mice. Additionally, BSE treatment decreased the levels of the pro-inflammatory cytokines TNF-a, IL-1β, and IL-6, and COX-2 in testis of LPS-induced endotoxemic mice models. In conclusion, BSE could be a potential nutraceutical for preventing the excessive immune related infertility.

Bovine mammary epithelial (MAC-T) cells are commonly used to study mammary gland development and mastitis. Lipopolysaccharide is a major bacterial cell membrane component that can induce inflammation. Autophagy is an important regulatory mechanism participating in the elimination of invading pathogens. In this study, we evaluated the mechanism underlying bacterial mastitis and mammary cell death following lipopolysaccharide treatment. After 24 h of 50 μg/mL lipopolysaccharide treatment, a significant decrease in the proliferation rate of MAC-T cells was observed. However, no changes were observed upon treatment of MAC-T cells with 10 μg/mL of lipopolysaccharide for up to 48 h. Thus, upon lipopolysaccharide treatment, MAC-T cells exhibit dose-dependent effects of growth inhibition at 10 μg/mL and death at 50 μg/mL. Treatment of MAC-T cells with 50 μg/mL lipopolysaccharide also induced the expression of autophagy-related genes ATG3, ATG5, ATG10, ATG12, MAP1LC3B, GABARAP-L2, and BECN1. The autophagy-related LC3A/B protein was also expressed in a dose-dependent manner upon lipopolysaccharide treatment. Based on these results, we suggest that a high dose of bacterial infection induces mammary epithelial cell death related to autophagy signals.

Periodontal disease is an inflammatory disease that affects the destruction of the bone supporting the tooth and connective tissues surrounding it. Periodontal ligament fibroblasts (PDLFs) induce overexpression of matrix metalloproteinase (MMP) involved in periodontal diseaseʼs inflammatory destruction. Osteoclasts take part in physiological bone remodeling, but they are also involved in bone destruction in many kinds of bone diseases, including osteoporosis and periodontal disease. This study examined the effect of baicalin on proteolytic enzymesʼ production and secretion of inflammatory cytokines in PDLFs and RAW 264.7 cells under the lipopolysaccharide (LPS)-induced inflammatory conditions. Baicalin inhibited the expression of the protein, MMP-1 and MMP-2, without affecting PDLFs’ cell viability, suggesting its possibility because of the inhibition of phosphorylation activation of mitogen-activated protein kinase’s p38, and the signal transduction process of nuclear factor κB (NFκB)-related protein. Also, baicalin reduced the expression of MMP-8 and MMP-9 in RAW 264.7 cells. This reduction is thought to be due to the inhibition of the signal transduction process of NFκB-related proteins affected by inhibiting p65RelA phosphorylation. Also, baicalin inhibited the secretion of nitric oxide and interleukin-6 induced by LPS in RAW 264.7 cells. These results suggest that baicalin inhibits connective tissue destruction in periodontal disease. The inhibition of periodontal tissue destruction may be a therapeutic strategy for treating inflammatory periodontal-diseased patients.

Baicalin is a flavonoid compound with many advantages, including anti-inflammatory agents and antioxidants. Lipopolysaccharide (LPS) is an endotoxin that induces neuronal damage through inflammatory response and oxidative stress reaction. This study was investigated the protective effects of baicalin on the oxidative stress and histopathological changes caused by LPS in hippocampus. Adult mice were divided into four groups; vehicle-treated, baicalin-treated, LPS-treated, and LPS and baicalin co-treated animals. Baicalin (10 mg/kg/day) and/or LPS (250 μg/kg/day) were intraperitoneally administered for seven consecutive days, and body weight was measured. Reactive oxygen species (ROS) level and lipid peroxidation level in the hippocampus were examined. Histopathological study was performed using hematoxylin and eosin staining manuals. LPS treatment decreased body weight and increased ROS and oxidative stress in the hippocampus. However, co-treatment with baicalin alleviated these changes caused by LPS. Severe histopathological changes were observed in the hippocampus of LPS-treated animals. Baicalin co-treatment attenuated the changes and preserved neuronal cells from LPS damage. These results showed that baicalin suppresses LPS-induced neuronal damage by alleviating oxidative stress in the hippocampus. Thus, this study demonstrated that baicalin exerts protective effects against LPS-induced oxidative stress in hippocampus.

Lysophosphatidic acid (LPA) is a lipid messenger mediated by G protein-coupled receptors (LPAR1-6). It is involved in the pathogenesis of certain chronic inflammatory and autoimmune diseases. In addition, it controls the self-renewal and differentiation of stem cells. Recent research has demonstrated the close relationship between periodontitis and various diseases in the human body. However, the precise role of LPA in the development of periodontitis has not been studied. We identified that LPAR1 was highly expressed in human periodontal ligament stem cells (PDLSCs). In periodontitis-mimicking conditions with Porphyromonas gingivalis -derived lipopolysaccharide (Pg-LPS) treatment, PDLSCs exhibited a considerable reduction in the cellular viability and osteogenic differentiation potential, in addition to an increase in the inflammatory responses including tumor necrosis factor-α and interleukin-1β expression and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation. Of the various LPAR antagonists, pre-treatment with AM095, an LPAR1 inhibitor, showed a positive effect on the restoration of cellular viability and osteogenic differentiation, accompanied by a decrease in NF-κB signaling, and action against Pg-LPS. These findings suggest that the modulation of LPAR1 activity will assist in checking the progression of periodontitis and in its treatment.

Lipopolysaccharide (LPS)는 염증유발 cytokine 분비를 자극하고 염증을 유발하는 그람음성균의 내독소이다. 본 연구에서는 LPS가 신경아교세포 활성과 해마에 있는 nuclear factor kappa B (NF-κB) 매개 염증유발 요소를 조절하는지를 조사하였다. 성체 수컷 쥐를 대조군과 LPS를 투여한 실험군으로 무작위로 나누어 vehicle 또는 LPS (250 μg/kg)를 5일 동안 복강투여하고 무게를 측정했다. 해마의 활성산소와 과산화지방질 수준을 분석하고, 형태학적 연구를 위해 Hematoxylin and eosin 염색을 시행하였다. 또한, 해마에서 ionized calcium-binding adapter molecule 1 (Iba-1), glial fibrillary acidic protein (GFAP), NF-κB, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α)의 발현을 확인하기 위해 Western blot 분석과 immunofluorescence 염색을 시행하였다. 그 결과, LPS를 투여한 쥐들의 체중이 감소하였다. LPS 투여는 활성산소와 과산화지방질 수준의 증가를 유발하였고 LPS를 투여한 쥐의 해마에서 심각한 조직병리학적 변화를 확인했다. 또한 LPS 투여는 신경교세포와 별아교세포의 표시물인 Iba-1과 GFAP의 발현을 증가시켰고, NF-κB의 발현과 IL-1β와 TNF-α와 같은 염증성인자의 발현을 증가시켰다. 이러한 결과들을 통해 LPS 투여는 해마손상과 염증반응을 유도한다는 것을 알 수 있고 LPS 투여가 해마조직에서 신경아교세포와 NF-κB에 매개된 염증인자들을 활성화시킨다는 것을 확인하였 다. 따라서, 본 연구는 LPS 투여는 해마조직에서 산화적 스트레스 증가와 염증인자 활성을 증가시켜 신경손상을 유도함을 보여준다.

Periodontal diseases have been associated with the development of cardiovascular diseases. Accumulating evidences have indicated that Porphyromonas gingivalis , a major periodontopathic pathogen, plays a critical role in the pathogenesis of atherosclerosis. In the present study, we demonstrated that P. gingivalis lipopolysaccharide (LPS) increases the mRNA and protein expression of matrix metalloproteinase-9 (MMP-9) in rat vascular smooth muscle cells. We showed that the MMP-9 expression induced by P. gingivalis LPS is mediated by the activation of signal transducer and activator of transcription 3 (STAT3) in vascular smooth muscle cells. Furthermore, the inhibition of STAT3 activity reduced P. gingivalis LPS-induced migration of vascular smooth muscle cells. Overall, our findings indicate that P. gingivalis LPS stimulates the migration of vascular smooth muscle cells via STAT3-mediated MMP-9 expression.

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.

The dynastid beetle Allomyrina dichotoma has been used as a herbal medicine. Recently, we performed de novo RNAsequencing of Allomyrina dichotoma and identified several antimicrobial peptide candidates based on in silico analysis.Among them, cationic antimicrobial peptide, Allomyrinasin, was selected and we assessed the anti-inflammatory activitiesof Allomyrinasin against mouse macrophage Raw264.7 cells. The results showed that Allomyrinasin decreased the nitricoxide production of the lipopolysaccharide-induced Raw264.7 cells. In addition, quantitative RT-PCR, ELISA and Westernblot analysis revealed that Allomyrinasin reduced cytokine expression levels in the Raw264.7 cells. Taken together, thesedata indicated that Allomyrinasin had anti-inflammatory activity in the lipopolysaccharide-induced Raw264.7 cells.

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.

Porphyromonas gingivalis, a foremost periodontal pathogen, has been known to cause periodontal diseases. Epidemiologic evidences have indicated the involvement of P. gingivalis in the development of cardiovascular diseases. In this study, we show that the P. gingivalis lipopolysaccharide increases the mRNA expression and protein secretion of interleukin-6 in vascular smooth muscle cells. We demonstrate that P. gingivalis LPS activates the extracellular signalregulated kinase 1/2 (ERK1/2), p38 mitogen-activated protein kinase (MAPK), and Akt, which mediate the IL-6 expression in vascular smooth muscle cells. Also, P. gingivalis LPS stimulates the vascular smooth muscle cell migration, which is a critical step for the progression of atherosclerosis. Moreover, neutralization of the IL-6 function inhibits the migration of vascular smooth muscle cells induced by P. gingivalis LPS. Taken together, these results indicate that P. gingivalis LPS promotes the expression of IL-6, which in turn increases the migration of vascular smooth muscle cells.

The bacterial lipopolysaccharide (LPS) mainly contributes to the structural integrity, survival and protection barrier against harsh environments. Therefore, the early stages in LPS or lipid A biosynthesis are attractive targets in the identification and development of inhibitors which would be effective against infections caused by Gram-negative bacteria. The bacterial outer membrane proteins (OMPs) meanwhile function as maintenance for structure, adhesion to other cells and substances, as well as development of resistance to antimicrobials. The LPS and LPS-related molecules, and OMPs are important immunogenic components of several important pathogens including Brucella, which have been extensively used in immunological studies and in the diagnosis of diseases. Here we review the importance, structure, functions and immunogenic aspects of LPS and OMPs particularly of Brucella which can be targeted for the prevention and diagnosis of brucellosis.