Safety evaluation of Gamisoyo-san: genotoxicity, acute toxicity, and influence on drug-metabolizing enzymes.

Gamisoyo-san is an herbal formula widely used to treat psychological issues, menopausal symptoms, and dysmenorrhea. However, there is insufficient information on its safety profile. This study aimed to confirm the genotoxic and acute toxic potential of Gamisoyo-san. We performed a battery of tests, which included a bacterial reverse mutation test (Ames test) using five bacterial strains, an in vitro chromosomal aberration test using Chinese hamster lung (CHL) cells, an in vivo micronucleus test in mice, and human Cytochrome P450 (CYP450) and UDP-glucuronosyltransferase (UGT) assays. In the acute toxicity study, male and female rats were orally administered Gamisoyo-san 1000, 2000, or 5000 mg/kg and observed for 14 days. The activities of human CYP450s and UGTs were evaluated using recombinant baculosomes. Gamisoyo-san showed no signs of genotoxicity in the five bacterial strains, CHL cells, or mouse bone marrow cells. The acute toxicity test showed that the median lethal dose (LD50) of Gamisoyo-san was greater than 5000 mg/kg in rats. Gamisoyo-san inhibited the activities of CYP1A2, CYP2C19, and UGT1A1. In conclusion, Gamisoyo-san may not exert severe toxicological events or genotoxic effects at doses up to 5000 mg/kg in rats.

Simultaneous Determination of Fourteen Marker Compounds in the Traditional Herbal Prescription, Geumgwesingihwan, Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry.

Geumgwesingihwan (GSH) is a traditional herbal prescription composed of eight medicinal herbs: Rehmannia glutinosa (Gaertn.) DC., Dioscorea japonica Thunb., Cornus officinalis Siebold and Zucc., Poria cocos Wolf, Paeonia suffruticosa Andrews, Alisma plantago-aquatica subsp. orientale (Sam.) Sam., Achyranthes bidentate Blume, and Plantago asiatica L. This study developed and validated an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method in the multiple reaction monitoring (MRM) mode for simultaneous determination of 14 compounds (allantoin, gallic acid, 5-(hydroxymethyl)furfural, geniposidic acid, oxypaeoniflorin, loganin, geniposide, paeoniflorin, ecdysterone, verbascoside, cornuside, benzoylpaeoniflorin, paeonol, and alisol B acetate) in GSH. The chromatographic separation of all marker analytes was carried out on an Acquity UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 µm) using gradient elution of a mobile phase of distilled water-acetonitrile containing 0.1% acetic acid. The newly established UPLC-MS/MS MRM method was validated by evaluating the linearity, the limits of detection and quantification, recovery, and precision. All markers were detected at concentrations of 6.94-4126.28 mg/kg. In addition, the recovery was 76.65-119.49% and the relative standard deviation value of the precision was 0.19-9.91%. The newly developed and validated UPLC-MS/MS assay will provide useful information for quality assessment of GSH.

In Vitro and In Vivo Genotoxicity Assessments and Phytochemical Analysis of the Traditional Herbal Prescription Siryung-Tang.

Siryung-tang (SRT) is a traditional herbal prescription containing Oryeong-san and Soshiho-tang that is used to treat digestive system diseases. We performed safety evaluations of SRT based on genotoxicity and developed an assay for quality control using high-performance liquid chromatography with a photodiode array detector. Genotoxicity was evaluated based on bacterial reverse mutation (Salmonella typhimurium TA1535, TA98, TA100, and TA1537, and Escherichia coli WP2 uvrA), chromosomal aberration (Chinese hamster lung cells), and micronucleus (mouse) tests. Quality control analysis was conducted using a SunFire C18 column and gradient elution with a distilled water-acetonitrile mobile phase system containing 0.1% (v/v) formic acid for 12 markers (5-(hydroxy-methyl)furfural, 3,4-dihydroxybenzaldehyde, liquiritin apioside, liquiritin, coumarin, baicalin, wogonoside, cinnamaldehyde, baicalein, glycyrrhizin, wogonin, and atractylenolide III). SRT showed no genotoxicity in three tests. Ames tests showed that SRT at 313-5000 µg/plate did not significantly increase the number of revertant colonies with or without metabolic activation among five bacterial strains. Moreover, in vivo micronucleus testing showed that SRT did not increase the frequency of bone marrow micronuclei. The number of chromosomal aberrations associated with SRT was similar to that observed in the negative controls. The 12 markers were detected at 0.04-16.86 mg/g in a freeze-dried SRT sample and completely eluted within 45 min. The extraction recovery was 95.39-104.319% and the relative standard deviation value of the precision was ≤2.09%. Our study will be used as basic data for the safety and standardization of SRT.

Anti-inflammatory effect of Gyeji-tang in a chronic obstructive pulmonary disease mouse model induced by cigarette smoke and lipopolysaccharide.

CONTEXT: Chronic obstructive pulmonary disease (COPD) is a chronic inflammatory lung disease associated with respiratory symptoms and narrowing of airways. Gyeji-tang (GJT) is a traditional Asian medicine that has been used to relieve early-stage cold symptoms, headache, and chills. OBJECTIVE: We examined the effect and potential molecular action mechanism of GJT in a mouse model of COPD induced by cigarette smoke (CS) plus lipopolysaccharide (LPS). MATERIALS AND METHODS: COPD was induced in C57BL/6J mice via daily exposure to CS for 1 h for 8 weeks and intranasal administration of LPS on weeks 1, 3, 5, and 7. GJT (100 or 200 mg/kg) or roflumilast (5 mg/kg) was administrated daily for the final 4 weeks of COPD induction. RESULTS: Administration of GJT significantly suppressed the CS/LPS-induced increases in: the numbers of total cells and macrophages in bronchoalveolar lavage fluid; the expression levels of tumour necrosis factor-α, interleukin (IL)-6, IL-1ß, and IL-8; the activities (phosphorylation) of nuclear factor kappa B and signal transducer and activator of transcription 3; and the expression levels of the structural remodelling markers, transforming growth factor beta, matrix metallopeptidase (MMP)-7, and MMP-9. DISCUSSION AND CONCLUSIONS: These results demonstrate that GJT prevents the lung inflammation and airway remodelling induced by CS plus LPS exposure in mice, suggesting that GJT may have therapeutic potential for the treatment of COPD.

Magnesium augments immunosuppressive effects of a corticosteroid in obese mice with airway inflammation.

BACKGROUND: Magnesium deficiency common in obesity is known to promote chronic low-grade inflammation and aggravate asthma symptoms; however, the effects of magnesium supplementation in obese asthmatic patients have not been investigated. OBJECTIVE: To examine the effects of magnesium co-administration with dexamethasone on airway inflammation in obese mice. METHODS: Female C57BL/6 mice were fed a high-fat diet, sensitized with ovalbumin (OVA) to induce allergic reactions, challenged with aerosolized OVA, and administered dexamethasone (3 mg/kg) with or without magnesium. Bronchial inflammation was analyzed based on the presence of inflammatory cells and cytokines in bronchoalveolar lavage fluid, total and OVA-specific IgE in serum, goblet cells ratios, bronchial wall thickness, and expression of α-smooth muscle actin. RESULTS: In obese mice, co-administration of magnesium and dexamethasone decreased IL-13 in bronchoalveolar lavage fluid and total and OVA-specific IgE in serum, and reduced α-smooth muscle actin-positive areas in the bronchi compared with mice treated with dexamethasone alone. However, no differences were observed in dexamethasone-treated normal-weight mice depending on magnesium supplementation. CONCLUSION: These results suggest that magnesium increases immunosuppressive effects of dexamethasone in airway inflammation aggravated by obesity, suggesting that magnesium supplementation may have a potential in alleviating asthma symptoms in obese patients with reduced responses to corticosteroids.

Safety assessment of Oryeong-san, a traditional herbal formula: Study of subacute toxicity and influence of cytochrome P450s and UDP-glucuronosyltransferases.

Oryeong-san is a traditional herbal formula that is used for the treatment of common genitourinary diseases in Korea and other Asian countries. However, little is known about its safety and influence on drug metabolism. In the present study, we investigated the subacute toxicity of an Oryeong-san water extract (OSWE) in rats and its effects on activities of drug-metabolizing enzymes. Subacute toxicity was modeled in animals exposed to treatment with the extract at multiple doses. Rats were given OSWE by oral gavage at 0, 1000, 2000 and 5000 mg/kg/day for 4 weeks. We checked general observations and investigated any changes of body/organ weight, food consumption, hematology, serum biochemistry, and urinalysis in vivo; and the activities of human microsomal cytochrome P450s (CYP450s) and UDP-glucuronosyltransferase (UGT) isozymes in vitro. We found that OSWE caused no significant toxicological changes at the doses tested. Therefore, the no observed adverse effect level of OSWE was more than 5000 mg/kg/day for male and female rats. OSWE inhibited the activities of CYP2C19 (IC50: 737.69 µg/mL) and CYP2E1 (IC50: 177.77 µg/mL). These results indicate that OSWE may be safe with no drug-related toxicity for up to 4 weeks and provide useful information concerning its potential to interact with conventional drugs or other herbal medicines.

The genotoxicity of an aqueous extract of Gyejibokryeong-hwan.

BACKGROUND: Gyejibokryeong-hwan (Guizhi Fuling Wan in China), a mixture of five herbal plants, is a well-known treatment for renal diseases including those associated with climacteric syndrome. However, the genotoxicity of Gyejibokryeong-hwan has not yet been well established. METHODS: The present study investigated that the genotoxicity of an aqueous extract of Gyejibokryeong-hwan (GJBRHE): an in vitro chromosomal aberration test using Chinese hamster lung cells, an in vitro bacterial reverse mutation assay (Ames test) with Salmonella typhimurium and Escherichia coli strains, and an in vivo micronucleus test using ICR mouse bone marrow. RESULTS: GJBRHE with or without the S9 mix showed no genotoxicity in the Ames test up to 5000 µg/plate or in the in vivo MN test up to 2000 mg/kg body weight. In contrast, the chromosomal aberration test showed that GJBRHE induced an increase in the number of chromosomal aberrations compared with the control after treatment for 6 h with 4200 µg/mL GJBRHE in the presence of the S9 mix and for 22 h with 800 µg/mL GJBRHE in the absence of the S9 mix. CONCLUSIONS: GJBRHE did not cause detectable genotoxic effects in the bacterial mutation test or the in vivo MN test, however genotoxic effect was detected in the in vitro chromosomal aberration assay. Our results suggest that GJBRHE may be associated with a low risk of carcinogenesis. Thus, further detailed experiments would be needed to clarify the compound responsible for inducing this genotoxicity of GJBRHE and to determine its mechanism.

Inhibitory effects of Ponciri Fructus on testosterone-induced benign prostatic hyperplasia in rats.

BACKGROUND: Benign prostatic hyperplasia (BPH) is non-cancerous condition of enlargement of the prostate, a common occurrence in older men. The immature fruits of Poncirus trifoliata (L.) Rafinesque (Rutaceae), Ponciri Fructus are widely used in traditional oriental medicine for the therapy of various diseases. However, little is known about the mechanism underlying the pathogenesis of BPH. In the present study, we investigated the protective effects of a Ponciri Fructus extract (PFE) on the development of BPH in a in a rat model of BPH induced by testosterone propionate (TP). METHODS: Male Sprague Dawley rats were used as a model of BPH after its induction by daily subcutaneous injections of TP/corn oil, for a period of four weeks. PFE was administrated daily 1 h before TP/corn oil injection by oral gavage at a dose level of 200 mg/kg during the 4 weeks of TP/corn oil injections. All rats were sacrificed at the end of the experiment, we measured the relative prostate weight, the levels of testosterone and dihydrotestosterone (DHT), histological changes, activities of antioxidant enzymes (catalase, glutathione peroxidase, glutathione reductase, and superoxide dismutase), and expression of proliferating cell nuclear antigen (PCNA). In addition, we also measured the inhibition (%) of 5α-reductase in the prostatic tissue. RESULTS: Our findings indicate that PFE significantly inhibited the development of BPH; decreased the relative prostate weight, the level of testosterone and DHT in serum and prostatic tissue, prostatic hyperplasia, expression of PCNA, and increased the antioxidant enzymes. Moreover, PFE showed a weak inhibitory activity on 5α-reductase. CONCLUSIONS: These results suggest that PFE may be used as a therapeutic agent for BPH via antiproliferative and antioxidant effects.

Genetically obese (ob/ob) mice are resistant to the lethal effects of thioacetamide hepatotoxicity.

Obesity increases the risk of chronic liver diseases, including viral hepatitis, alcohol-induced liver disease, and non-alcoholic steatohepatitis. In this study, we investigated the effects of obesity in acute hepatic failure using a murine model of thioacetamide (TA)-induced liver injury. Genetically obese ob/ob mice, together with non-obese ob/+ littermates, were subjected to a single intraperitoneal injection of TA, and examined for signs of hepatic injury. ob/ob mice showed a significantly higher survival rate, lower levels of serum alanine aminotransferase and aspartate aminotransferase, and less hepatic necrosis and apoptosis, compared with ob/+ mice. In addition, ob/ob mice exhibited significantly lower levels of malondialdehyde and significantly higher levels of glutathione and antioxidant enzyme activities compared with their ob/+ counterparts. Bioactivation analyses revealed reduced plasma clearance of TA and covalent binding of [(14)C]TA to liver macromolecules in ob/ob mice. Together, these data demonstrate that genetically obese mice are resistant to TA-induced acute liver injury through diminished bioactivation of TA and antioxidant effects.

Anti-inflammatory effects of Salvia plebeia R. Br extract in vitro and in ovalbumin-induced mouse model.

BACKGROUND: Asthma is an increasing global health problem, and novel strategies to prevent or ameliorate the condition are needed. Here, the effects of 80 % ethanol extracts of Salvia plebeia R. Br. (SE) on an induced inflammatory response were investigated. RESULTS: Salvia plebeia R. Br. inhibited production of pro-inflammatory cytokines, such as TNF-α and IL-6, as well as nitric oxide (NO) in LPS-stimulated RAW 264.7 cells. NO and pro-inflammatory cytokine production was suppressed more effectively by SE of the aerial parts (SE-A) than of the roots (SE-R) of S. plebeia. In BEAS-2B cells, both SE-A and SE-R inhibited the increase in production of the inflammatory cytokines IL-6 and IL-8. We also investigated the anti-asthmatic effects of SE in an ovalbumin (OVA)-induced BALB/c mouse model. SE-A treatment significantly reduced the number of airway eosinophils, IL-4 and IL-13 levels, mucus production, and inflammatory infiltration, as compared with the corresponding levels in the untreated, OVA-induced mice, and had similar effects to dexamethasone. CONCLUSIONS: Salvia plebeia ethanol extract ameliorated the induced inflammatory response in RAW 264.7 and BEAS-2B cells, with more effective inhibition noted for SE-A than for SE-R. SE-A treatment was effective in improving the histopathological changes in the lungs of asthma model mice via modulation of eosinophils and Th2 cytokines. These results suggest that SE-A can be considered as a therapeutic agent that can potentially relieve asthma.

Acute and subacute toxicity of an ethanolic extract of Melandrii Herba in Crl:CD sprague dawley rats and cytotoxicity of the extract in vitro.

BACKGROUND: Melandrii Herba, a medicinal plant, has been used in Korea for treatment of bacterial and fungal infection. However, the safety and toxicity of Melandrii Herba have not yet been established. Therefore, we investigated the acute and subacute toxicity of an ethanolic extract of Melandrii Herba (MHEE) in Crl:CD Sprague Dawley rats and cytotoxicity of MHEE in vitro. METHODS: To study acute toxicity, rats were treated with MHEE at single doses of 0, 500, 1000, and 2000 mg/kg administered by oral gavage, and body weight, clinical signs, and mortality were observed after dosing. To study subacute toxicity, rats were treated with MHEE at doses of 0, 500, 1000, and 2000 mg/kg administered once a day by gavage for 4 weeks. We measured clinical signs, mortality, gross pathological findings, body and organ weights, food consumption, serum biochemistry, and conducted hematology and urinalysis. The cytotoxicity of MHEE was assayed by measuring the viability of prostate cell lines including normal prostate stromal WPMY-1, normal prostate epithelial RWPE-1, and benign prostatic hyperplasia epithelial BPH-1 cells at various concentrations of MHEE in vitro. RESULTS: Single oral doses of MHEE caused no significant difference in rat clinical signs, mortality, or body weight. The lethal dose of MHEE was considered to be >2000 mg/kg. Daily oral doses of MHEE for 4 weeks did not result in any significant changes in rat mortality, gross pathological findings, relative organ weights, food consumption, hematology, serum biochemistry, or urinalysis. At MHEE >1000 mg/kg/day, salivation was increased in both male and female rats. However, the salivation caused by the MHEE treatment was not accompanied by pathological changes in body weight or gross pathological findings, and we considered the salivation as a minor symptom. Therefore, no adverse effects were seen at 2000 mg/kg/day or less. MHEE showed no cytotoxic effects on either normal prostate or benign prostatic hyperplasia cell lines. CONCLUSIONS: Administration of MHEE in Crl:CD Spradgue Dawley rats is nontoxic and is safe for at least a month.

Gyeji-tang water extract exerts anti-inflammatory activity through inhibition of ERK and NF-κB pathways in lipopolysaccharide-stimulated RAW 264.7 cells.

BACKGROUND: Gyeji-tang (GJT, Guizhi Tang in Chinese, Keishi-to in Japanese) is a traditional herbal decoction composed of 5 medicinal herbs. GJT has been used to treat the common cold, headaches, and fever in Asian countries including Korea, China, and Japan. In the present study, we investigated the inhibitory effect of a water extract of GJT on inflammatory response using the murine macrophage cell line, RAW 264.7. METHODS: RAW 264.7 macrophages were treated with lipopolysaccharide (LPS) to upregulate inflammatory genes. Cells were pretreated with various concentrations of GJT for 4 h and stimulated with LPS for an additional 20 h. Productions of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), cyclooxygenase-2 (COX-2), and prostaglandin E2 (PGE2) were measured by enzyme-linked immunosorbent assays (ELISAs). Protein expressions of heme oxygenase (HO)-1, extracellular signal-regulated kinase (ERK), and nuclear factor kappa-B (NF-κB) were analyzed by immunoblotting. RESULTS: Treatment with the GJT extract enhanced expression of HO-1 in macrophages without cytotoxicity. GJT extract significantly inhibited proinflammatory cytokines TNF-α and IL-6 in LPS-stimulated cells. GJT suppressed LPS-induced COX-2 expression, leading to a decrease in COX-2-derived PGE2 level. In addition, GJT extract prevented phosphorylation of ERK and NF-κB translocalization to the nucleus in LPS-treated RAW 264.7 cells. CONCLUSION: These data suggest that GJT has anti-inflammatory possibly through blocking ERK and NF-κB signaling pathways.

Role of Klotho, an antiaging protein, in pulmonary fibrosis.

Klotho is a recently discovered antiaging protein. Although many researchers are investigating the roles of Klotho in chronic kidney diseases and cancer, however, there are no studies on the roles of Klotho in chronic pulmonary diseases. The purpose of this study was to define the role of Klotho in pulmonary fibrosis using a murine model of ovalbumin (OVA)-induced chronic asthma and in BEAS-2B human bronchial epithelial cells. In an in vivo experiment, mice were sensitized by intraperitoneal injection of OVA (20 µg/mouse), followed 1 week later by an airway challenge with 1 % OVA solution delivered three times a week for 4 weeks. In an in vitro experiment, we investigated the effects of stimulated with interleukin (IL)-4 and tumor necrosis factor (TNF)-α on Klotho protein and VEGF and transforming growth factor (TGF)-ß1/Smad3 signaling in BEAS-2B cells. Klotho decreased and VEGF and TGF-ß1 levels increased with increasing duration of OVA challenge. Similar findings were found for the expression of these proteins in lung tissue. The collagen content in lung tissue increased with repeated OVA challenge. In the in vitro experiment, Klotho expression decreased and VEGF and TGF-ß1/Smad3 expression increased after IL-4 (50 ng/mL) and TNF-α (50 ng/mL) stimulation. Pretreatment with 25, 50, and 100 ng/mL of Klotho protein significantly attenuated the increases in VEGF and TGF-ß1/Smad3 expression levels after IL-4 and TNF-α treatment, and reduced α-smooth muscle actin expression in concentration-dependent manner. Klotho protein inhibited the fibrotic response by suppressing VEGF and TGF-ß1/Smad3 expression. These results suggest that Klotho protein may be crucial to inhibiting fibrosis associated with chronic airway diseases.

Safety assessment of Guibi-tang: Subchronic toxicity study in Crl:CD SD rats.

Guibi-tang (Kihi-To in Japanese and Qui-Pi-Tang in Chinese) is a multiherbal traditional Korean medicinal formula used for treatment of amnesia, fatigue, poor memory or forgetfulness, anemia, insomnia, and necrosis. The aim of the present study was to investigate potential safety, if any, of subchronic administration of Guibi-tang aqueous extract (GBT) in laboratory animals. For this study, 0, 1000, 2000, and 5000 mg/kg/day of GBT was administered to Crl:CD Sprague Dawley rats (10/sex/group) for 13 weeks via oral gavage. Administration of the GBT did not result in any mortality, body weight, food consumption, hematology, serum biochemistry, clinical or ophthalmological signs or changes in urinalysis, gross findings, or organ weight. However, histopathology revealed forestomach hyperplasia and duodenum mucosal hyperplasia in rats of both sexes at the highest dose of GBT, 5000 mg/kg/day. Therefore, the no-observed-adverse-effect level in rats was established for GBT at 2000 mg/kg/day under the conditions of this study.

Genotoxicity evaluation of Oryeong-san water extract using in vitro and in vivo tests.

BACKGROUND: Oryeong-san, a mixture of five herbal plants, is a well-known therapy for renal-associated diseases such as those manifesting edema, dysuria, and oliguria. METHODS: In the present study, we investigatee the potential genotoxic effects of a water extract of Oryeong-san (ORSE) in three mutagenicity assays (an in vitro bacterial reverse mutation assay (Ames test) with Salmonella typhimurium and Escherichia coli strains, an in vitro mammalian chromosomal aberration test using Chinese hamster lung cells, and an in vivo micronucleus test using ICR mice bone marrow). RESULTS: ORSE showed no genotoxicity in the Ames test up to 5000 µg/plate; the in vitro chromosome aberration test showed no significant structural aberrations with and without the S9 mix up to 5000 µg/mL, or the in vivo micronucleus test up to 2000 mg/kg body weight. CONCLUSIONS: In conclusion, under the current test conditions, ORSE seems safe for use; however, other genotoxicity tests (e.g. sister-chromatid exchange or Comet) or chronic toxicity tests are warranted.

Samsoeum water extract attenuates allergic airway inflammation via modulation of Th1/Th2 cytokines and decrease of iNOS expression in asthmatic mice.

BACKGROUND: Samsoeum has long been used in Korea and other Asian countries as a traditional medicine to treat various diseases. In the present study, we investigated the antiasthma effect of the herbal medicine Samsoeum water extract (SSEW) using an in vivo ovalbumin (OVA)-induced asthmatic model. METHODS: Female BALB/c mice were sensitized by an intraperitoneal injection of OVA and subsequently challenged with nebulized OVA. We investigated the number of inflammatory cells, the production of Th1/Th2 cytokines and chemokine in bronchoalveolar lavage fluid (BALF), histological changes in lung tissue, the infiltration of inflammatory cells and hyperplasia of goblet cells in lung tissue, the levels of immunoglobulin E (IgE) in BALF and plasma, and the expression of inducible nitric oxide synthase (iNOS) in lung tissue. RESULTS: Our findings indicated that SSEW decreased the accumulation of inflammatory cells (particularly, eosinophil and neutrophil) and regulated the balance in the production of Th1/Th2 cytokines and chemokine in BALF. Moreover, SSEW suppressed the level of IgE in BALF and plasma, and inhibited the infiltration of inflammatory cells, hyperplasia of goblet cells, and the expression of iNOS in lung tissue. CONCLUSIONS: Collectively, these results suggest that, because of its anti-inflammatory and antiasthma properties, SSEW may be useful in reducing airway inflammation in the treatment of allergic asthma.

Evaluation of a water extract of So-Cheong-Ryong-Tang for acute toxicity and genotoxicity using in vitro and in vivo tests.

BACKGROUND: So-Cheong-Ryong-Tang, a traditional Korean medicine, has been used empirically for the treatment of asthma, allergic rhinitis, and colds for hundreds of years. However, its genotoxicity has been rarely examined. METHODS: We therefore investigated the genotoxicity of an aqueous extract of So-Cheong-Ryong-Tang (SCRT) in two in vitro and one in vivo assays: a bacterial reverse mutation assay (Ames test), a chromosomal aberration assay, and a micronucleus assay, respectively. RESULTS: In the bacterial reverse mutation assay, SCRT did not increase revertant colony numbers in Salmonella. typhimurium strains (TA98, TA100, TA1535, and TA1537) or an Escherichia coli strain (WP2 uvrA) regardless of metabolic activation or the duration of treatment. However, statistically significant differences in the incidence of chromosomal aberrations following SCRT>4000 µg/mL were observed in Chinese hamster lung cells exposed with or without an S9 enzyme and cofactor mixture. CONCLUSIONS: These results suggest further genotoxic testing of SCRT, such as a comet assay, to ascertain its generally recognized safety.

Soshiho-tang water extract inhibits ovalbumin-induced airway inflammation via the regulation of heme oxygenase-1.

BACKGROUND: Soshiho-tang, known as Xio-hai-Hu-Tang in Chinese and Sho-Saiko-to in Japanese, has been widely used as a therapeutic agent. Its pharmacological effects include anti-inflammatory, antioxidant, antihepatic fibrosis, antitumor and immunomodulating activities. However, little is known regarding its effects on allergic asthma. Therefore, the aim of the present study was to investigate whether the Soshiho-tang water extract (SSTW) has antiasthmatic effects on airway inflammation in an ovalbumin (OVA)-induced mouse model. METHODS: BALB/c mice were used as a model of asthma after induction by sensitization and challenge with OVA. We measured change in eosinophils, other inflammatory cells, and T helper 2 (Th2)-type cytokines, such as interleukin (IL)-4, IL-5, IL-13, IL-17, IL-33, and chemokine (eotaxin) in bronchoalveolar lavage fluid (BALF), presence of total and OVA-specific immunoglobulin (Ig)E in plasma, and expression of mucus production and heme oxygenase (HO)-1 protein in lung tissue. RESULTS: Our results show that SSTW had a suppressive effect on eosinophil influx into BALF and decreased the levels of Th2-type cytokines. Moreover, SSTW exhibited a marked decrease in mucus hypersecretion, total and OVA-specific IgE levels, and significantly induced HO-1 protein expression. CONCLUSIONS: These results suggest that SSTW may be used as a valuable therapeutic agent for treating various inflammatory diseases including allergic asthma.

Luffa cylindrica suppresses development of Dermatophagoides farinae-induced atopic dermatitis-like skin lesions in Nc/Nga mice.

CONTEXT: The fruit pulp of Luffa cylindrica Roemer (Cucurbitaceae) (LC) has been used to induce hemostasis, resolve phlegm and clear fever in traditional Korean medicine. However, the efficacy of LC has not been examined in atopic dermatitis (AD). OBJECTIVE: A 70% ethanol extract of LC was evaluated to determine anti-inflammation and anti-AD effects in vitro and in vivo. MATERIALS AND METHODS: The inhibitory effects of LC on the production of PGE2 and histamine were respectively measured in lipopolysaccharide-treated (1 µg/mL) RAW264.7 macrophages and phorbol-12 myristate 13-acetate (50 nM) and A23187 (1 µM)-stimulated HMC-1 mast cells. The production of AD-related chemokines (RANTES, TARC, and MDC) were evaluated in IFN-γ and TNF-α-stimulated (10 ng/mL, each) HaCaT keratinocytes. LC (10 mg/mouse/d) was topically applied to the dorsal skin and ears of Dermatophagoides farina (Pyroglyphidae)-sensitized Nc/Nga mice for 4 weeks. RESULTS: The IC50 values of LC on PGE2 and histamine production were 16.89 and 139.9 µg/mL, individually. The production of TARC and RANTES were inhibited 20% and 12% by LC (50 µg/mL) in HaCaT cells, respectively (p < 0.05). In sensitized-NC/Nga mice, the plasma levels of IgE and histamine were suppressed 36% and 41% by LC, respectively (p < 0.05). LC also reduced hemorrhage, hypertrophy, and hyperkeratosis of the epidermis and infiltration of mast cells in the dorsal skin and ear. DISCUSSION AND CONCLUSION: LC can inhibit AD-like skin lesions and reduce the generation of IgE via inhibition of the inflammatory responses. LC has potential as a therapeutic agent to treat allergic diseases, including AD.

Effects of maternal exposure to di(2-ethylhexyl)phthalate (DEHP) during pregnancy on susceptibility to neonatal asthma.

Di(2-ethylhexyl) phthalate (DEHP) is used as a plasticizer and is widely dispersed in the environment. In this study, we investigated the effects of maternal exposure to DEHP during pregnancy on neonatal asthma susceptibility using a murine model of asthma induced by ovalbumin (OVA). Pregnant BALB/c mice received DEHP from gestation day 13 to lactation day 21. Their offspring were sensitized on postnatal days (PNDs) 9 and 15 by intraperitoneal injection of 0.5µg OVA with 200µg aluminum hydroxide. On PNDs 22, 23 and 24, live pups received an airway challenge of OVA for 30min. Offspring from pregnant mice that received DEHP showed reductions in inflammatory cell count, interleukin (IL)-4, IL-13, and eotaxin in their bronchoalveolar lavage fluid and in total immunoglobulin E and OVA-specific IgE in their plasma compared with offspring from pregnant mice that did not receive DEHP treatment. These results were consistent with histological analysis and immunoblotting. Maternal exposure to DEHP reduces airway inflammation and mucus production in offspring, with a decrease in inducible nitric oxide synthase (iNOS) in the lung tissue. This study suggests that maternal exposure to DEHP during pregnancy reduces asthmatic responses induced by OVA challenge in offspring. These effects were considered to be closely related to the suppression of Th2 immune responses and iNOS expression.