Activation of transient receptor potential vanilloid 3 by the methanolic extract of Schisandra chinensis fruit and its chemical constituent γ-schisandrin.

Transient receptor potential vanilloid 3 (TRPV3) is a non-selective cation channel with modest permeability to calcium ions. It is involved in intracellular calcium signaling and is therefore important in processes such as thermal sensation, skin barrier formation, and wound healing. TRPV3 was initially proposed as a warm temperature sensor. It is activated by synthetic small-molecule chemicals and plant-derived natural compounds such as camphor and eugenol. Schisandra chinensis (Turcz.) Baill (SC) has diverse pharmacological properties including antiallergic, anti-inflammatory, and wound healing activities. It is extensively used as an oriental herbal medicine for the treatment of various diseases. In this study, we investigated whether SC fruit extracts and seed oil, as well as four compounds isolated from the fruit can activate the TRPV3 channel. By performing whole-cell patch clamp recording in HEK293T cells overexpressing TRPV3, we found that the methanolic extract of SC fruit has an agonistic effect on the TRPV3 channel. Furthermore, electrophysiological analysis revealed that γ-schisandrin, one of the isolated compounds, activated TRPV3 at a concentration of 30 µM. In addition, γ-schisandrin (~100 µM) increased cytoplasmic Ca2+ concentrations by approximately 20% in response to TRPV3 activation. This is the first report to indicate that SC extract and γ-schisandrin can modulate the TRPV3 channel. This report also suggests a mechanism by which γ-schisandrin acts as a therapeutic agent against TRPV3-related diseases.

Can We Determine Sasang Constitutional Body Type Merely by Facial Inspection?

OBJECTIVES: This study aimed to assess the inter-observer concordance rate of anthroscopic examination on facial features among experts in Sasang constitutional medicine (SCM) in order to evaluate the presence of statistical differences in facial structural characteristics among different body types of Sasang constitution (SC), and to develop an objective method for facial analysis for diagnosing SC types to prevent SCM experts from misdiagnosis by their perceptional errors about faces. METHODS: This was a double-blinded cross-sectional study conducted on 174 people's faces. Ten SCM experts participated in this study. Frontal and lateral photographs of subjects were standardized and displayed to 10 SCM experts for diagnosing the SC type by anthroscopic examination alone (experiment 1). The subjects' faces were analyzed by photogrammetric method to investigate the presence of any typical structural characteristics of the faces to differentiate SC type (experiment 2). Comparing subjects' SC type with anthroscopic diagnosis by 10 SCM experts, the inter-observer concordance rates were measured (experiment 1). Using photogrammetric facial analysis, a multinomial logistic model was made for analyzing the correlation of SC type and subjects' facial structural configuration (experiment 2). RESULTS: The inter-observer concordance rate of anthroscopic examination was 2.9% in experiment 1. Using a multinomial logistic fitting model, the predicted probability for determining SC type was 52.8-57.6% in experiment 2 (p < 0.05). Prototype composite faces were also created from photographs of subjects who received the same SC type from the SCM experts. CONCLUSIONS: As SC type cannot be precisely diagnosed using anthroscopic examination alone, SCM needs a definitive objective and scientific diagnosing method to be a scientifically verified alternative medicine and be globalized in future.

Spirodela polyrhiza extract modulates the activation of atopic dermatitis-related ion channels, Orai1 and TRPV3, and inhibits mast cell degranulation.

CONTEXT: Spirodela polyrhiza (L.) Schleid. (Lemnaceae), Spirodelae Herba (SH), has been known to relieve inflammation, urticaria and skin symptoms including pruritus, eczema and rash. OBJECTIVE: The effects of SH extract on two calcium ion channels, Orai1 and TRPV3, and their potential as novel therapeutics for atopic dermatitis (AD) were investigated. The regulatory role of Orai1 on mast cell degranulation was evaluated. MATERIALS AND METHODS: The dried leaves of SH were extracted by 70% methanol. Effects of SH extract (100 µg/mL) in an HEK293T cell line overexpressing human Orai1 or TRPV3 were assessed. Ion channel modulation in transfected HEK293T cells was measured using a conventional whole-cell patch-clamp technique. IgE-antigen complex-stimulated mast cell degranulation was measured by ß-hexosaminidase assay with morphological observation after treatment with 20, 50 and 100 µg/mL SH extract. RESULTS: SH extract (100 µg/mL) significantly inhibited Orai1 activity (63.8 ± 0.97%) in Orai1-STIM1 co-overexpressed HEK293T cells. SH extract significantly increased TRPV3 activity (81.29 ± 0.05% at -100 mV) compared with the positive control 2-APB (100 µM), which induced full activation. SH extract inhibited degranulation in IgE-antigen complex-stimulated RBL-2H3 mast cells by decreasing ß-hexosaminidase activity (3.14 ± 0.03, 2.56 ± 0.12 and 2.29 ± 0.08 mU/mg, respectively). CONCLUSION: Our results suggested that SH extract could treat abnormal skin barrier pathologies in AD through modulation of the activities of the calcium ion channels Orai1 and TRPV3 and inhibition of mast cell degranulation. This is the first report of an herbal effect on the modulation of ion channels associated with skin barrier disruption in AD pathogenesis.

Effects of the Fruit Extract of Tribulus terrestris on Skin Inflammation in Mice with Oxazolone-Induced Atopic Dermatitis through Regulation of Calcium Channels, Orai-1 and TRPV3, and Mast Cell Activation.

ETHNOPHARMACOLOGICAL RELEVANCE: In this study, we investigated the effects of Tribulus terrestris fruit (Leguminosae, Tribuli Fructus, TF) extract on oxazolone-induced atopic dermatitis in mice. MATERIALS AND METHODS: TF extract was prepared with 30% ethanol as solvent. The 1% TF extract with or without 0.1% HC was applied to the back skin daily for 24 days. RESULTS: 1% TF extract with 0.1% HC improved AD symptoms and reduced TEWL and symptom scores in AD mice. 1% TF extract with 0.1% HC inhibited skin inflammation through decrease in inflammatory cells infiltration as well as inhibition of Orai-1 expression in skin tissues. TF extract inhibited Orai-1 activity in Orai-1-STIM1 cooverexpressing HEK293T cells but increased TRPV3 activity in TRPV3-overexpressing HEK293T cells. TF extract decreased ß-hexosaminidase release in RBL-2H3 cells. CONCLUSIONS: The present study demonstrates that the topical application of TF extract improves skin inflammation in AD mice, and the mechanism for this effect appears to be related to the modulation of calcium channels and mast cell activation. This outcome suggests that the combination of TF and steroids could be a more effective and safe approach for AD treatment.

The extract of Cinnamomum cassia twigs inhibits adipocyte differentiation via activation of the insulin signaling pathway in 3T3-L1 preadipocytes.

CONTEXT: Obesity is associated with a number of diseases with metabolic abnormalities such as type 2 diabetes (T2D). Medicinal plants have been widely used for the treatment of obesity and related complications. OBJECTIVE: In this study, we investigated the antidiabetic properties of the extract of twigs of Cinnamomum cassia Blume (Lauraceae) (Cinnamomi Ramulus; CR) in 3T3-L1 murine preadipocytes. MATERIALS AND METHODS: 3T3-L1 cells were differentiated into adipocytes for 3 d in insulin-conditioned medium and then treated with CR extract at concentrations of 100 and 500 µg/mL for 6 d. Adipocyte differentiation was measured by Oil Red O staining, and the expression of master transcription factors, peroxisome proliferator-activated receptor-gamma (PPARγ), CCAAT/enhancer binding protein-alpha (C/EBPα), and sterol regulatory element binding protein-1c (SREBP-1c), and lipid metabolism factors were investigated by reverse transcription-polymerase chain reaction (RT-PCR). The activation of the AMP-activated protein kinase (AMPK)/insulin signaling pathway was assessed by western blot analysis. RESULTS: CR extract significantly reduced lipid accumulation and down-regulated the expression of PPARγ, C/EBPα, and SREBP-1c in 3T3-L1 adipocytes. CR extract also suppressed the expression of fatty acid synthase (FAS), acyl-CoA synthase, and perilipin. Moreover, CR extract markedly up-regulated the phosphorylation of AMPK and acetyl-CoA carboxylase (ACC). In addition, CR extract effectively increased the expression levels of glucose transporter-4 (GLUT-4), phosphatidylinositol 3-kinase (PI3K), and insulin receptor substrate-1 (IRS-1) in 3T3-L1 adipocytes. DISCUSSION AND CONCLUSION: These results suggest that CR extract may have therapeutic potential as a natural agent for the improvement of T2D via regulation of the insulin-dependent signaling pathway.

Comparison of Gut Microbiota between Sasang Constitutions.

The Sasang constitutional medicine has long been applied to diagnose and treat patients with various diseases. Studies have been conducted for establishment of scientific evidence supporting Sasang Constitutional (SC) diagnosis. Recent human microbiome studies have demonstrated individual variations of gut microbiota which can be dependent on lifestyle and health conditions. We hypothesized that gut microbial similarities and discrepancies may exist across SC types. We compared the difference of gut microbiota among three constitutions (So-Yang, So-Eum, and Tae-Eum), along with the investigation of anthropometric and biochemical parameters. Firmicutes and Bacteroidetes were predominant phyla in all SC types. The median plot analysis suggested that Firmicutes and Bacteroidetes appeared more abundant in SE and TE, respectively, in the male subjects of 20-29 years old. At the genus level, Bifidobacterium and Bacteroides manifested the difference between SE and TE types. For anthropometry, body weight, body mass index, and waist circumference of the TE type were significantly higher than those of the other types. Overall, findings indicated a possible link between SC types and gut microbiota within a narrow age range. Further investigations are deemed necessary to elucidate the influences of age, gender, and other factors in the context of SC types and gut microbiota.

Wen-pi-tang-Hab-Wu-ling-san, a Polyherbal Medicine, Attenuates ER Stress in 3T3-L1 Preadipocytes by Promoting the Insulin Signaling Pathway.

The endoplasmic reticulum (ER) is an organelle that functions to synthesize, fold, and transport proteins. ER stress is a key link between type 2 diabetes (T2D), obesity, and insulin resistance. In this study, we investigated the effect of WHW on the ER stress response and the insulin signaling pathway in 3T3-L1 preadipocytes. 3T3-L1 preadipocytes were differentiated into adipocytes, and ER stress was then induced by treatment with tunicamycin. ER stress-induced adipocytes were treated with different concentrations of WHW for 24 h. The expression of ER stress-related molecules such as X-box-binding protein-1 (XBP-1), glucose-regulated protein 78 (GRP78), C/EBP-homologous protein 10 (CHOP10), and eukaryotic initiation factor 2 α (eIF2 α ) and signaling molecules such as phosphatidylinositol 3-kinase (PI3K), insulin receptor substrates-1 (IRS-1), and c-Jun N-terminal protein kinase (JNK) were investigated. WHW significantly inhibited the expression of XBP-1, GRP78, CHOP10, and eIF2 α in ER stress-induced 3T3-L1 adipocytes. WHW also increased the PI3K expression and the IRS-1 phosphorylation but decreased the phosphorylation of JNK in ER stress-induced 3T3-L1 adipocytes. Our results indicate that WHW inhibits ER stress in adipocytes by suppressing the expression of ER stress-mediated molecules and the insulin signaling pathway, suggesting that WHW may be an attractive therapeutic agent for managing T2D.

Anti-diabetic effect of Wen-pi-tang-Hab-Wu-ling-san extract in streptozotocin-induced diabetic rats.

OBJECTIVES: Wen-pi-tang-Hab-Wu-ling-san (WHW) is an oriental herbal prescription formulated using 14 herbs and has been used to cure chronic renal failure in Korean oriental medicine. In this study, we investigated the anti-diabetic effect of WHW in the streptozotocin-induced diabetic rats. MATERIALS AND METHODS: Diabetes was induced by streptozotocin (STZ, 60 mg/kg, i.p.) in rats. WHW extract (100 mg/kg) was orally dosed once a day for four weeks. The results were compared with standard antidiabetic drug, glibenclamide (3 mg/kg, p.o). RESULTS: Significant decrease in body weight and insulin levels and increase in blood glucose, triglycerides, urea nitrogen (BUN), and creatinine were detected in STZ-induced diabetic rats with disruption and disappearance of pancreatic and kidney cells and decrease in insulin producing beta cells. However, these diabetic changes were significantly inhibited by treatment with WHW extract. In the oral glucose tolerance test, the extract produced a significant decrease in glycemia 60 minutes after the glucose pulse. CONCLUSIONS: Based on these results, we suggest that WHW extract has favorable effects in protecting the STZ-induced hyperglycemia, renal damage, and beta-cell damage in rats.

The antioxidant effects of Joongpoongtang 05 on brain injury after transient focal cerebral ischemia in rats.

Traditional Korean medicines with antioxidant properties are believed to have potential therapeutic effects against oxidative stress in cerebral ischemia. In this study, the antioxidant effects of Joongpoongtang 05 (JP05) were evaluated in rat middle cerebral artery occlusion (MCAO). Male Sprague-Dawley rats treated with JP05 (100 and 200 mg/kg body weight, p.o.) for 5 days showed a reduction in infarct sizes and a decrease in oxidants after MCAO. Furthermore, a significant inhibition of enzymatic antioxidants in the ischemic cerebral cortex was observed. Treatment with JP05 reversed the inhibition to normal levels. In the glutathione system, the reduced glutathione (GSH) level was decreased while oxidized glutathione (GSSG) was increased in MCAO rats. Treatment with JP05 increased the GSH/GSSG ratio and redox index by enhancing the synthesis of GSH from GSSG. Moreover, in vitro studies showed that JP05 has strong antioxidant activity in the Trolox equivalent antioxidant capacity, oxygen radical absorbance capacity and 1,1-diphenyl-2-picrylhydrazyl radical assays. The present findings suggest that JP05 treatment can contribute to a protective effect on brain ischemia through antioxidant mechanisms.