Jinju Kim

Extracts from Citrus unshiu promote immune-mediated inhibition of tumor growth in a murine renal cell carcinoma model.

AIM OF THIS STUDY Citrus unshiu (Satsuma mandarin, SM) is a citrus fruit the peel of which has been used as a traditional Chinese medicine to treat common cold, relieve exhaustion, and cancer. In this study, we examined how effectively the content and peel extracts of SM can suppress cancer growth. The mechanism underlying cancer-suppressing properties of SM was investigated in tumor-bearing mice with renal carcinoma cell, Renca. MATERIALS AND METHODS Effectiveness of SM in tumor suppression was evaluated by measuring size of tumor mass in tumor-bearing mice treated with various doses of SM content and peel extracts. Proliferation of tumor cells and splenocytes was determined by MTT assay and [³H]TdR uptake, respectively. Relevant immunological mechanisms were chased by assaying cytokines including TGF-β, IL-6, IFN-γ, and TNF-α by ELISA. RESULTS The content and peel extracts of SM inhibited the growth of tumor cells in tumor-bearing mice. Especially, average tumor volume of two groups treated with 3 and 30 mg peel extracts per mouse weight (kg) were significantly decreased to 52.32% (p<0.05) and 68.72% (p<0.01), respectively. To identify tumor regression mechanism, anti-tumor cytokines measured in Con A-activated splenocytes from tumor-bearing mice. IFN-γ was increased in both of the peel extract-treated groups, while TNF-α, which had been decreased by tumor growth, was rescued to the normal level in SM content and peel extracts-treated groups. However, SM content and peel extracts did not inhibit proliferation and tumor-proliferative cytokines including TGF-β and IL-6 production of tumor cells. CONCLUSION These results indicate that SM content and peel extracts have anti-tumor properties in the tumor-bearing murine model. The mechanism underlying the anti-tumor effects of SM extracts is strongly suggested to be via boosting cytokines such as IFN-γ and TNF-α, enhancing immune-mediated anti-tumor properties.

Extract of Salvia miltiorrhiza (Danshen) induces Nrf2-mediated heme oxygenase-1 expression as a cytoprotective action in RAW 264.7 macrophages

ETHNOPHARMACOLOGICAL RELEVANCE Danshen (Salvia miltiorrhiza) is widely used in traditional herbal medicines for relief of a variety of symptoms related to complications arising from vascular diseases such as hypertension, diabetes, and atherosclerosis. Induction of heme oxygenase-1 (HO-1) expression protects against oxidative stress-induced cell damage, which plays an important role in cytoprotection in a variety of pathological models. MATERIALS AND METHODS In the present study, we investigated the effect of Danshen on the up-regulation of HO-1, an inducible and cytoprotective enzyme in RAW 264.7 macrophages. Molecular mechanisms underlying the effects, especially protective effects, was elucidated by analyzing the activation of transcription factors and their upstream signalling, and by evaluating the inhibitory effect of HO-1 on ROS production. RESULTS Danshen induced HO-1 mRNA expression and protein production, and nuclear translocation of NF-E2-related factor 2 in RAW 264.7 macrophages. Pharmacological inhibitors of PI3K/Akt and MEK1 attenuated HO-1 induction in Danshen-stimulated RAW 264.7 macrophages. Furthermore, Danshen pretreatment reduced intracellular production of reactive oxygen species after stimulation with hydrogen peroxide; this effect was reversed by the HO-1 inhibitor ZnPP. CONCLUSION Danshen induced HO-1 expression through PI3K/Akt-MEK1-Nrf2 pathway and reduced intracellular production of reactive oxygen species via induction of HO-1 expression. The results support a role of HO-1 in the cytoprotective effect of Danshen.

Lilium lancifolium Thunb. extract attenuates pulmonary inflammation and air space enlargement in a cigarette smoke-exposed mouse model

ETHNOPHARMACOLOGICAL RELEVANCE Lilium lancifolium Thunb. (Liliaceae) has long been used as a traditional medicine in Korea and China to treat bronchitis, pneumonia, and other pulmonary ailments. AIM OF THE STUDY Cigarette smoke (CS) is a major risk factor for the development of pulmonary inflammatory response; it also triggers pulmonary alveoli enlargement. In the present study, we investigate the effects of Lilium lancifolium Thunb. root extract on pulmonary inflammatory responses in a CS-exposed mouse model. MATERIALS AND METHODS Water extract of Lilium lancifolium Thunb. root was fed to C57BL/6 mice prior CS exposure every day for 3 weeks. The numbers of macrophages and neutrophils in bronchoalveolar lavage fluid (BALF) were counted. The relative inflammatory factors, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-1 beta (IL-1β), monocyte chemotactic protein-1 (MCP-1), and matrix metalloproteinase-12 (MMP-12) were measured by real-time PCR, ELISA, or Western blot analysis. The average alveoli size was determined by lung histology. RESULTS Lilium lancifolium Thunb. root extract was found to significantly inhibit the numbers of macrophages and neutrophils in BALF due to CS exposure. Lilium lancifolium Thunb. root extract also reduced the protein secretion levels of TNF-α, IL-6, IL-1β, and MCP-1 in BALF and the RNA expression levels of TNF-α, IL-6, IL-1β, MCP-1, and MMP-12 in lung tissue compared with mice only exposed to CS. Moreover, MMP-12 in serum was down regulated in Lilium lancifolium Thunb. root extract treated mice compared with CS-exposed mice. Finally, a morphometric analysis of the lungs of Lilium lancifolium Thunb. root extract treated mice demonstrated a significant reduction in airspace size compared to mice only exposed to CS. CONCLUSION Our results show that Lilium lancifolium Thunb. root extract reduces lung inflammation and airspace enlargement in a CS-exposed mouse model. These data indicate that Lilium lancifolium Thunb. root extract is a therapeutic candidate for pulmonary inflammation and emphysema caused by CS.

Methyl Gallate Exhibits Potent Antitumor Activities by Inhibiting Tumor Infiltration of CD4+CD25+ Regulatory T Cells

CD4+CD25+ regulatory T (Treg) cells play crucial roles in the host response to tumors. Increasing evidence supports the existence of elevated numbers of Treg cells in solid tumors and hematologic malignancies. In this study, the effects of methyl gallate on Treg cells were examined. Methyl gallate inhibited Treg cell-suppressive effects on effector CD4+ T cells and Treg migration toward tumor environment. The expression of Treg surface markers including CTLA-4, CCR4, CXCR4, and glucocorticoid-induced TNFR was significantly suppressed upon methyl gallate treatment. Furthermore, forkhead box P3 (Foxp3) expression was also significantly decreased by methyl gallate, suggesting that the suppressive effects of methyl gallate on Treg were medicated by decrease of Treg-specific transcription factor Foxp3. In tumor-bearing hosts, methyl gallate treatment substantially reduced tumor growth and prolonged the survival rate. In contrast, nu/nu mice did not show decreased tumor progression in response to methyl gallate. In addition, in tumor-bearing Treg-depleted mice, tumor growth and the survival rates were not changed by methyl gallate treatment, strongly suggesting that the main therapeutic target of methyl gallate in tumor suppression was related to modulation of the CD4+CD25+ Treg cell functions. In the spleen of tumor-bearing mice, methyl gallate treatment induced a significant decrease in the CD4+CD25+Foxp3high Treg cell population. Especially, the number of tumor-infiltrating CD25+Foxp3high Treg cells was significantly lower in methyl gallate-treated mice. These results suggest that methyl gallate can be used to reverse immune suppression and as a potentially useful adjunct for enhancing the efficacy of immune-based cancer therapy.

Effects of Schisandra chinensis Baillon (Schizandraceae) on lipopolysaccharide induced lung inflammation in mice

ETHNOPHARMACOLOGICAL RELEVANCE Schisandra chinensis Baillon (Sc), an anti-inflammatory herb that has been used in traditional Chinese medicine for thousands of years, is frequently used to treat upper respiratory tract infections. AIM OF THE STUDY This study was conducted to evaluate the ability of a water extract of Sc to prevent airway inflammation both in vitro and in vivo. MATERIALS AND METHODS Human lung alveolar epithelial-derived A549 cells were stimulated with to interleukin-1β, tumor necrosis factor-α, and interferon-γ (IL-1β, TNF-α, and INF-γ; cytokine mixture; CM) and treated with Sc extracts. They were then evaluated using nitric oxide (NO), IL-8 and monocyte chemotactic protein-1 (MCP-1) secretions. In the in vivo study, BALB/c mice were challenged with lipopolysaccharide (LPS) to induce acute airway inflammation. After this challenge, the mice were treated with Sc extracts (10, 50 and 100mg/kg) by oral administration, and inflammatory cells in the bronchoalveolar lavage (BAL) fluid were counted. IL-6 and TNF-α secretions were measured using an enzyme-linked immunosorbent assay. Lung tissues of the LPS treated mice were prepared and stained with hematoxylin and eosin (HE) for histological examination. RESULTS In the A549 cells, Sc extracts dose-dependently and significantly inhibited CM-induced NO production and reduced IL-8 and MCP-1 secretions. Sc extracts efficiently suppressed neutrophil and macrophage infiltrations of lung tissues and increased IL-6 and TNF-α levels in BAL fluid in LPS-instilled BALB/c mice. In addition, Sc extracts treatment inhibited pathologic progress in the lung tissues, as confirmed by H&E staining. These findings indicate that Sc extracts could be potentially useful for the treatment of acute lung inflammation and acute lung injury.

Bambusae Caulis in Taeniam extract reduces ovalbumin-induced airway inflammation and T helper 2 responses in mice

ETHNOPHARMACOLOGICAL RELEVANCE Bambusae Caulis in Taeniam (BC) was known as traditional herbal medicine with anti-inflammatory property in the Orient. AIM OF THE STUDY Allergic asthma is inflammatory disease of airways associated with enhanced T helper (Th) 2 lymphocytes responses to allergens, leading to eosinophilic infiltration and elevated serum IgE levels. Although there were some studies that BC extract had an anti-inflammatory property, there was no study on asthma. In present study, we investigated the suitability of BC extract as a therapeutic candidate in the treatment of allergic airway disease in ovalbumin-induced asthma model. MATERIALS AND METHODS Balb/C mice (female, 6 weeks old) were treated by ovalbumin sensitization and nebulization, and used as asthma model. The number of eosinophil in bronchoalveolar lavage (BAL) fluid and the degree of eosinophila were investigated by hematoxylin and eosin stain and the infiltration of inflammatory cells into lung tissues was examined by staining by hematoxylin and eosin solution. The levels of interleukin (IL)-4 in BAL fluid, immunoglobulin E (IgE) in serum, interferon (IFN)-gamma and IL-4 production in splenocyte culture from Balb/C mice (not treated, 6 weeks old) that incubated with or without BC extract for 48 h were determined by enzyme-linked immunosorbent assay. RESULTS The level of eosinophils was decreased by treatment of the animals with BC extract (40 mg/kg) and correspondingly, a significantly lowered degree of eosinophila was also reported (p<0.01). In lung tissue, BC extract reduced the increased immune cell infiltration induced by OVA (p<0.05). Furthermore, the levels of IL-4 and IgE in BAL fluid or serum up-regulated by OVA was decreased by BC extract. Finally, IFN-gamma production was significantly increased (p<0.01), while IL-4 production significantly decreased (p<0.01), after treatment of the culture supernatants of splenocytes with BC extract. CONCLUSIONS These results indicated that BC extract reduces OVA-induced airway inflammation and Th 2 response in mice, suggesting that BC extract can be a therapeutic candidate for allergic airway disease, including asthma.

Inhibitory effect of Agrimoniae Herba on lipopolysaccharide-induced nitric oxide and proinflammatory cytokine production in BV2 microglial cells

Abstract Objectives: Agrimoniae Herba has been used as an anti-inflammatory agent in traditional medicine. Nitric oxide and proinflammatory cytokines produced by activated microglia may be a possible etiological factor of neurodegenerative disorders. We evaluated whether Agrimoniae Herba could have an anti-inflammatory effect on lipopolysaccharide-induced BV2 microglial cells. Methods: The effects of Agrimoniae Herba on lipopolysaccharide-induced nitric oxide and proinflammatory cytokine production in BV2 microglial cells were evaluated by nitric oxide assay, enzyme-linked immunosorbent assay and western blotting. Results: Agrimoniae Herba had no cytotoxicity and suppressed lipopolysaccharide-induced nitric oxide production in BV2 microglial cells. Agrimoniae Herba also suppressed lipopolysaccharide-induced production of proinflammatory cytokines such as tumor necrosis factor, interleukin 1 beta and interleukin 6 in a dose-dependent manner. Agrimoniae Herba inhibited the expression of inducible nitric oxide synthase. Discussion: Taken together, these findings indicate that Agrimoniae Herba may be used as a form of pharmaceutical acupuncture therapy in the treatment of brain inflammation.

Topical Application of Angelica sinensis Improves Pruritus and Skin Inflammation in Mice with Atopic Dermatitis-Like Symptoms

Angelica sinensis (AS) is one of the most popular medicinal foods used as a hematopoietic herb and also traditionally applied topically for skin disorders. However, the effectiveness of AS on atopic dermatitis (AD) has not been reported yet. This study was conducted to evaluate the antipruritic and anti-inflammatory effects of AS on regulating AD-related mediators in DNCB (2,4-dinitrochlorobenzene)-induced mice. AS was topically applied to the dorsal skin of DNCB-challenged mice for 11 days. Alteration of skin thickness was measured for assessment of histological improvement. In addition, the number of mast cells, the level of serum immunoglobulin E (IgE), the counting of scratching behavior, and the expression of substance P were evaluated. Also, the expressions of cytokines, nuclear factor κB (NF-κB), phospho-IκBα, and mitogen-activated protein kinases (MAPKs) were measured for evaluating the improvement of skin inflammation. The repeated treatment of AS significantly inhibited the skin thickness, the number of mast cells, and the level of serum IgE. Moreover, AS significantly suppressed the increased scratching behavior and the expression of substance P compared to the DNCB group. Topical application of AS also reduced the level of cytokines (IL-4, IL-6, TNF-α, and IFN-γ) as well as the expressions of NF-κB, phospho-IκBα, and phospho-MAPKs in the dorsal skin. The results of our study suggest that topical application of AS might have efficacy for modulating pruritus and inflammation in AD. Further studies are required to further characterize the mechanism of actions of AS.

Stemona tuberosa prevented inflammation by suppressing the recruitment and the activation of macrophages in vivo and in vitro

ETHNOPHARMACOLOGICAL RELEVANCE Stemona tuberosa (ST) is a traditional herbal medicine used for the treatment of various respiratory diseases in eastern Asia. AIM OF THE STUDY We investigated the anti-inflammatory effects of a ST water extract in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and in cigarette smoke (CS)-induced lung inflammation mouse models. MATERIALS AND METHODS RAW 264.7 macrophages were treated with the ST extract and stimulated by LPS. The expressions of pro-inflammatory mediators were evaluated by using nitric oxide (NO) assay, enzyme-linked immunosorbent assay and Western blot analysis. After the C57BL/6 mice were exposed to CS, they were administrated with the ST extract. The accumulated inflammatory cells in the bronchoalveolar lavage fluid (BALF) were counted. Also, real-time polymerase chain reaction and hematoxylin and eosin staining were performed in lung tissues. RESULTS The ST extract treatment reduced the production of NO via blocking the expressions of cyclooxygenase-2 and inducible nitric oxide synthase protein in RAW 264.7 macrophages. In addition, ST extract treatment decreased the secretions of inflammatory cytokines and regulated NF-κB activation by inhibiting the phosphorylation of IκB and the mitogen-activated protein kinase pathway. Also, ST extract administration to mice reduced the infiltrations of macrophages into BALF and the histological inflammatory changes in lung tissues. Furthermore, administration of the ST extract regulated the levels of tumor necrosis factor-α, interleukin (IL)-6, IL-1β, monocyte chemoattractant protein-1 and matrix metalloproteinases-12 in the lungs. CONCLUSION These findings suggested that ST extract attenuated pulmonary inflammatory responses by inhibiting the expression of diverse inflammatory mediators in vivo and in vitro.

Schizonepeta tenuifolia inhibits the development of atopic dermatitis in mice.

Historically, Schizonepeta tenuifolia (ST) has been used for the treatment of skin disorders, such as allergic dermatitis, eczema, and inflammatory diseases. In this study, we examined whether ST inhibited 2,4‐dinitrochlorobenzene (DNCB)‐induced atopic dermatitis (AD) in BALB/c mice. In histopathological analyses of the epidermis and dermis, skin thickness was significantly increased in DNCB‐induced mice as compared with normal group. Treatment with ST inhibited this inflammatory change and markedly suppressed the secretion of immunoglobulin E, tumor necrosis factor α, and interleukin 6 levels in the serum of DNCB‐induced mice. In addition, ST treatment significantly restored the upregulation of proinflammatory factors, such as nuclear factor (NF)‐κB and mitogen‐activated protein kinase expression. Taken together, due to its ability to suppress inflammatory factors and upregulate proinflammatory factors, ST may be useful as a therapeutic treatment for AD. ST extract application decreased both epidermis and dermis thickness in DNCB‐induced mice. In serum, ST reduced immunoglobulin E, tumor necrosis factor, and interleukin 6 level. In addition, ST suppressed NF‐κB activation as well as the mitogen‐activated protein kinase activities. Copyright