Antiproteinuric Effects of Green Tea Extract on Tacrolimus-Induced Nephrotoxicity in Mice.

INTRODUCTION: It has been reported that proteinuria is an early predictive marker in detection of tacrolimus (TAC) nephrotoxicity. The aim of this study was to investigate the antiproteinuric effects of green tea extract (GTE) on TAC-induced acute nephrotoxicity in mice. METHODS: The mice (n = 20) were divided into 4 groups (n = 5 per group); control group mice were intraperitoneally (IP) injected with 0.9% saline, TAC group mice were IP injected with TAC 1 mg/kg, and inducible nitric oxide synthase (iNOS) inhibitor group mice were given in addition NG-nitro-L-arginine-methyl ester 12 mmol/L by subcutaneous injection. TAC-GTE group mice were given TAC by IP injection and GTE 100 mg/kg by subcutaneous injection. RESULTS: The 24-hour urine protein amounts were significantly increased in TAC group mice (36.1 ± 9.9 mg/d) compared with control group mice (13.3 ± 5.4 mg/d) and significantly decreased in TAC-GTE group mice (19.1 ± 6.9 mg/d, P < .01) compared with TAC group mice. The nitric oxide (NO) production by TAC was significantly suppressed by GTE and iNOS inhibitor injection. Renal tissue malondialdehyde (MDA) level was significantly increased in the TAC group compared with the control group and was significantly decreased in the TAC-GTE group compared with that of the TAC group. The antioxidant enzyme activities of superoxide dismutase and catalase were significantly suppressed in the TAC group compared with the control group and were restored in the GTE injection group. CONCLUSIONS: GTE treatment has beneficial antiproteinuric effects on TAC-induced acute renal injury in mice.

The antiproteinuric effects of green tea extract on acute cyclosporine-induced nephrotoxicity in rats.

BACKGROUND: It has been reported that the proteinuria is an early useful marker to detect cyclosporine (CsA) nephrotoxicity. The aim of this study was to investigate the antiproteinuric effects of green tea extract (GTE) on CsA-induced acute renal injury in rats. METHODS: The rats (n = 28) were divided into four groups (n = 7/group); controls intraperitoneally (IP) injected with 0.9% saline; CsA group IP injected CsA (50 mg/kg); inducible nitric oxide synthase (iNOS) inhibitor group administered in addition NG-nitro-L-arginine-methyl ester (12 mmol/L) subcutaneously and CsA-GTE group of CsA IP plus GTE (100 mg/kg) subcutaneously. RESULTS: The 24-hour urine proteins were significantly increased among the CsA (22.6 ± 3.1 mg/d) compared with the control (7.1 ± 1.5 mg/d) and significantly decreased in the CsA-GTE group (8.2 ± 1.8 mg/d, P < .01). Nitric oxide production induces by CsA treatment was significantly suppressed by GTE and iNOS inhibitor. Renal tissue malondialdehyde level was significantly increased in the CsA compared with controls and significantly decreased in the CsA-GTE group. The antioxidant enzyme activities of superoxide dysmutase and catalase, which were significantly suppressed in the CsA compared with the control group, were restored in the CsA-GTE cohort. CONCLUSION: GTE treatment of rats showed meaningful antiproteinuric effects through antioxidative activity in kidneys from CsA-induced acute renal injury.

Procoagulant and prothrombotic effects of the herbal medicine, Dipsacus asper and its active ingredient, dipsacus saponin C, on human platelets.

BACKGROUND: In spite of the growing popularity of herbal medicines and natural food supplements, their effects on cardiovascular homeostasis remain largely unknown, especially regarding pro-thrombotic risks. OBJECTIVE: In the present study, 21 herbal tea extracts were screened for the procoagulant activities on platelets, an important promoter of thrombosis to examine if herbal medicines or natural products may have prothrombotic risks. We discovered that Dipsacus asper (DA), known to have analgesic and anti-inflammatory effects, potently induced procoagulant activities in platelets. We tried to identify the active ingredient and elucidate the underlying mechanism. RESULTS: Among 10 major ingredients of DA, dipsacus saponin C (DSC) was identified as a key active ingredient in DA-induced procoagulant activities. DSC-induced procoagulant activities were achieved by the exposure of phosphatidylserine (PS) and PS-bearing microparticle generation that were caused by the alteration in the activities of phospholipid translocases: scramblase and flippase. These events were initiated by increased intracellular calcium and ATP depletion. Notably, DSC induced a series of apoptotic events including the disruption of mitochondrial membrane potential, translocation of Bax and Bak, cytochrome c release and caspase-3 activation. The key roles of apoptotic pathway and caspase activation were demonstrated by the reversal of DSC-induced PS exposure and procoagulant activities with the pretreatment of caspase inhibitors. Interestingly, EGTA reversed DSC-induced procoagulant activities and apoptotic events suggesting that an intracellular calcium increase may play a central role. These results were also confirmed in vivo where platelets of the rats exposed to DSC or DA exhibited PS exposure. Most importantly, DSC or DA administration led to increased thrombus formation. CONCLUSION: These results demonstrate that herbal medicines or natural products such as DA or DSC might have prothrombotic risks through procoagulant activation of platelets.

High-dose squalene ingestion increases type I procollagen and decreases ultraviolet-induced DNA damage in human skin in vivo but is associated with transient adverse effects.

BACKGROUND: Evidence for beneficial effects of squalene on ultraviolet (UV)-induced photoageing of the skin is lacking. AIM: To investigate whether squalene supplementation improves signs and molecular markers of photoageing in human skin in vivo. METHODS: In total, 40 female volunteers aged > 50 years received two different doses [13.5 g/day (low-dose group) and 27 g/day (high-dose group)] of squalene for 90 days. At baseline and at the completion of the study, facial wrinkles were measured using skin replicas. Skin samples were taken to compare type I procollagen and matrix metalloproteinase 1 mRNA levels by real-time reverse-transcriptase PCR, and for type I procollagen immunostaining. Skin samples were also taken 24 h after 2 x minimal erythema dose (MED) of UV irradiation before and after squalene intake to assess UV-induced thymine dimer formation and keratinocytic apoptosis. RESULTS: In total, 37 subjects completed the trial. Transient loose stool was experienced by 35% of volunteers in the low-dose group and 55% in the high-dose group. Facial wrinkles decreased significantly (P < 0.05) in the high-dose group, while procollagen type I mRNA levels and MED increased significantly in the low-dose group. Procollagen immunostaining tended to increase in both groups. Facial erythema decreased and pigmentation increased significantly in both groups. UV-induced keratinocytic apoptosis and thymine dimer staining were substantially reduced in both groups. CONCLUSIONS: Daily ingestion of 13.5 or 27 g of squalene per day resulted in antiageing effects in photoaged skin. However, in view of the frequent incidence of loose stool experienced by the subjects, the risk-benefit ratio of high-dose squalene supplementation is too high to recommend it for treating skin ageing.

Apoptotic effect of hesperidin through caspase3 activation in human colon cancer cells, SNU-C4.

Hesperidin, a known flavonoid constituent of citrus, reduces the proliferation of many cancer cells. The apoptotic effects of hesperidin on human colon cancer cells, SNU-C4, were determined at concentrations of 1-100 microM. At 100 microM, hesperidin reduced cell viability to 65.00+/-0.05% of control values in a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell death induced by hesperidin showed apoptotic features in 4,6-diamidino-2-phenylindole (DAPI) staining and in terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assays. Examination of the expression of apoptosis-regulating genes indicated that hesperidin treatment decreased the expression of B-cell CLL/lymphoma 2 (BCL2) mRNA, and increased the expression of BCL2-associated X protein (BAX). The expression and activity of the major apoptotic factor caspase3 (CASP3) was increased significantly with hesperidin treatment. Hesperidin down-regulated the protein expression of pro-CASP3, and up-regulated the level of active CASP3. Thus, these results suggest that hesperidin could induce apoptosis in human colon cancer cells through CASP3 activation.

Human hair growth enhancement in vitro by green tea epigallocatechin-3-gallate (EGCG).

Green tea is a popular worldwide beverage, and its potential beneficial effects such as anti-cancer and anti-oxidant properties are believed to be mediated by epigallocatechin-3-gallate (EGCG), a major constituent of polyphenols. Recently, it was reported that EGCG might be useful in the prevention or treatment of androgenetic alopecia by selectively inhibiting 5alpha-reductase activity. However, no report has been issued to date on the effect of EGCG on human hair growth. This study was undertaken to measure the effect of EGCG on hair growth in vitro and to investigate its effect on human dermal papilla cells (DPCs) in vivo and in vitro. EGCG promoted hair growth in hair follicles ex vivo culture and the proliferation of cultured DPCs. The growth stimulation of DPCs by EGCG in vitro may be mediated through the upregulations of phosphorylated Erk and Akt and by an increase in the ratio of Bcl-2/Bax ratio. Similar results were also obtained in in vivo dermal papillae of human scalps. Thus, we suggest that EGCG stimulates human hair growth through these dual proliferative and anti-apoptotic effects on DPCs.

The effect of erythrodiol-3-acetate on the expressions of matrix metalloproteinase-1 and type-1 procollagen caused by ultraviolet irradiated cultured primary old aged human skin fibroblasts.

Methanol and aqueous extracts of Styrax japonica used traditionally for the treatments of skin elastic materials were screened in vitro for the matrix metalloproteinase (MMP)-1 inhibitor actions. The methylene chloride soluble fraction of methanol extract from the stems of S. japonica showed significant MMP-1 inhibition in primary old aged human skin fibroblasts caused by ultraviolet (UV) irradiation. Main triterpenoids were isolated by repeated column chromatography. Among them, the triterpenoid erythrodiol-3-acetate reduced the expression of MMP-1 and induced the expression of type-1 procollagen at the protein levels in a dose-dependent manner caused by UV irradiated cultured old aged human skin fibroblasts. Taken together, our results suggest that erythrodiol-3-acetate plays an important role in the skin aging process caused by UV irradiation.

Coptis japonica root extract induces apoptosis through caspase3 activation in SNU-668 human gastric cancer cells.

Apoptosis-modulating approaches offer an attractive opportunity for therapeutic use for many tumors. We investigated the effects of the roots of Coptis japonica var. dissecta (Ranunculaceae) on human gastric cancer cells, SNU-668. The cytotoxicity of Coptis japonica at 100 microg/ml (methanol extract) by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was 13.89 +/- 1.91% of control value. Considering the features by 4,6-diamidino-2-phenylindole (DAPI) staining and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, it was confirmed that the death of SNU-668 cells was due to apoptosis. In the apoptosis-regulating genes, BCL2 expression was diminished out, whereas BAX and CASP3 expressions were increased, compared with control. Furthermore, the activity of caspase3 was significantly increased by Coptis japonica treatment. These results suggest that Coptis japonica could induce apoptotic anticancer effect through caspase3 activation on SNU-668 human gastric cancer cells.

Effects of Paeonia lactiflora root extracts on the secretions of monocyte chemotactic protein-1 and -3 in human nasal fibroblasts.

In the field of Oriental medicine, the root of Paeonia lactiflora (Paeoniae Radix) has been prescribed usually to treat the common cold, and it was thought to alleviate upper respiratory infection or nasal inflammation. Monocyte chemotactic proteins (MCP)-1 and MCP-3 are known as the most potent chemokines to mediate allergic inflammation. The object of the study was to investigate the effect of Paeoniae Radix on the release of the chemokines such as MCP-1 and MCP-3. To detect the secretion of MCPs, an enzyme-linked immunosorbent assay was performed for human nasal mucosal fibroblasts after stimulation with several cytokines such as tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) and interleukin-1beta (IL-1beta). The secretion of MCP-1 was increased after stimulation with TNF-alpha or IL-1beta treatment. Co-treatment of TNF-alpha and IFN-gamma, or IL-1beta and IFN-gamma increased the secretion of MCP-1 and MCP-3. Paeoniae Radix was tested with inflammatory cytokines, and the results showed that the Paeoniae Radix significantly decreased the secretion of MCP-1 and MCP-3. In our study, Paeoniae Radix may play an important role in nasal inflammation with a modulation of MCP-1 and MCP-3.

Effects of Jaoga-Yukmiwon(R), a Korean herbal medicine, on chondrocyte proliferation and longitudinal bone growth in adolescent male rats.

Externally visible growth of the body is the result of proliferation of chondrocytes and longitudinal bone growth. The effects of the Korean herbal medicine, Jaoga-Yukmiwon((R)), on the growth of adolescent rats were investigated in the present study. The proliferation ratio of chondrocytes was calculated from 5-bromo-2'-deoxyuridine incorporation during DNA synthesis. The 5-bromo-2'-deoxyuridine incorporation ratio of the control group was 21.1 +/- 3.5%, and Jaoga-Yukmiwon((R)) administration markedly increased the ratio to 34.5 +/- 4.6% (p < 0.05). The bone formation rate of longitudinal bone was estimated by labelling with tetracycline, which binds to newly formed bone. The bone formation rate in the tibia of Jaoga-Yukmiwon((R))-administered rats was increased to 405.1 +/- 4.3 micro m (p < 0.05) from 292.2 +/- 11.8 micrometer (control value). The height of the growth plate in the administrated rats was increased to 614.6 +/- 10.0 micro m (p < 0.05) from 552.3 +/- 17.1 micro m. The bone morphogenetic protein-2 immunostaining in the growth plate was also increased. These results suggest that Jaoga-Yukmiwon((R)) may promote longitudinal bone growth during the developmental period.

Auricular acupuncture decreases neuropeptide Y expression in the hypothalamus of food-deprived Sprague-Dawley rats.

The aim of the present study was to investigate whether stimulation of auricular acupuncture point has any effects on the expression of neuropeptide Y (NPY), appetite-inducing factor particularly abundant in the mammalian hypothalamus. In food-deprived condition, enhanced NPY expression was detected in both the arcuate nucleus (ARN) and the paraventricular nucleus (PVN) of the hypothalamus via immunohistochemistry in Sprague-Dawley rats. Needling the unfed rats on the auricular point resulted in decreased NPY levels in both the ARN and PVN, while it increased NPY levels in the ARN and PVN of fed rats. The present findings indicate that auricular acupuncture may affect NPY expression in the ARN and PVN of the hypothalamus.

The use of Q-switched Nd:YAG laser in the treatment of superficial pigmented lesions in Koreans.

AIM: There are still insufficient clinical reports concerning quality-switched Nd:YAG laser (QSNYL) in the treatment of superficial pigmented lesions in Asians. The purpose of this study was to analyse the efficacy and side effect profiles of QSNYL-assisted pigment removal in brown skin. METHODS: A total of 71 patients, presenting a wide gamut of superficial epidermal lesions, were treated with QSNYL. Clinical responses were assessed by comparing photographs that were taken serially in every treatment. RESULTS: Treatment using the QSNYL is reported individually for the various superficial pigmented lesions in the skin of Korean patients. CONCLUSION: QSNYL may be a beneficial alternative tool for the treatment of a number of benign pigmented lesions including freckles, lentigines and unilateral lentiginosis.

Clinical use of the Q-switched Nd:YAG laser for the treatment of acquired bilateral nevus of Ota-like macules (ABNOMs) in Koreans.

BACKGROUND: There are still insufficient clinical reports about quality-switched Nd:YAG laser (QSNYL) in the treatment of acquired bilateral nevus of Otalike macules (ABNOMs) in Asians. OBJECTIVE: To analyze the efficacy and side-effect profiles of QSNYL treatment of ABNOMs in Korean skin. METHODS: A prospective study was designed to follow 10 Korean patients with ABNOMs through laser treatment until maximal eradication of the lesions had been achieved. RESULTS: Five patients (50%) with ABNOMs were treated with excellent or good results. The more treatments a patient underwent, the greater the possibility of improvement. There were no cases of persistent skin textural change or persistent erythema. CONCLUSION: The clinical data support QSNYL being a beneficial alternative tool for treating ABNOMs in brown skin. This is, to the best of the authors' knowledge, the first report about laser treatment of ABNOMs using QSNNL.

Protective effects of puerariaeflos against ethanol-induced apoptosis on human neuroblastoma cell line SK-N-MC.

Puerariaeflos (PF) is an oriental medical herb for alcohol abuse. To investigate whether PF possesses protective effects against ethanol (EtOH)-induced cytotoxicity in the central nervous system, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometric analysis, DNA fragmentation assay, and reverse transcription-polymerase chain reaction were performed on SK-N-MC human neuroblastoma cells. Cells treated with EtOH exhibited several apoptotic features, while those pre-treated with PF prior to EtOH exposure showed a decreased occurrence of apoptotic features. In addition, PF pre-treatment inhibited the EtOH-induced increase in caspase-3 mRNA expression. These results suggest that PF may exert protective effects against EtOH-induced apoptosis in human neuroblastoma cells.

Auricular acupuncture increases cell proliferation in the dentate gyrus of Sprague-Dawley rats.

In a previous study, we have reported that acupuncture increases cell proliferation in the dentate gyrus after transient global ischemia in gerbils. In the present study, the effect of auricular acupuncture (AA) on dentate cell proliferation was examined in appropriately fed and food-deprived adult Sprague-Dawley (S-D) rats. 5-Bromo-2'-deoxyuridine-5'-monophosphate (BrdU) immunohistochemistry revealed a significant increase in cell birth in the dentate gyrus of both groups after AA treatment. In addition, the effect of AA on cell proliferation in the dentate gyrus was more profound in the food-supplied group. These findings demonstrate that AA enhances cell proliferation in the dentate gyrus of adult rats.

Acupuncture increases cell proliferation in dentate gyrus after transient global ischemia in gerbils.

The effects of acupuncture on cell proliferation in the dentate gyrus of gerbils after transient global ischemia were investigated in this study. Acupuncture was performed on Zusanli (ST36), which is a well known acupoint in animals and humans. In Oriental medicine, Zusanli has been commonly used for the enhancement of functional recovery in stroke patients. Through 5-bromo-2'-deoxyuridine (BrdU) immunohistochemistry, an increase in cell birth in the dentate gyrus of gerbils after ischemic injury was detected. Interestingly, acupunctural treatment in ischemic gerbils resulted in a significant increase in the number of BrdU-positive cells in the dentate gyrus. The present findings indicate that acupuncture may affect cell proliferation in the dentate gyrus of gerbils after ischemic injury.

Differential expression of nicotinamide adenine dinucleotide phosphate-diaphorase in hypothalamic areas of obese Zucker rats.

Several studies have suggested that the activity of nitric oxide synthase (NOS) may be involved in the regulation of food intake in the genetically obese Zucker rats. In the present study, we investigated the expression of NOS in various hypothalamic regions of obese and lean Zucker rats using nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry. Obese Zucker rats showed significantly lower staining intensities of NADPH-diaphorase-positive neurons in the paraventricular nucleus (PVN), lateral hypothalamic area (LHA) and ventromedial hypothalamic nucleus (VMH) than lean Zucker rats did. The differences in staining intensities between obese and lean Zucker rats were large in both the PVN and LHA, but such differences were relatively small in the VMH.