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Stimulatory Effects of Ginsan on the Proliferation and Viability of Mouse Spleen Cells.

The Korean Journal of Physiology and Pharmacology.2010;14(3):133-137. doi:10.4196/kjpp.2010.14.3.133

Ginsan is an acidic polysaccharide purified from Panax ginseng, a famous oriental herb. Although a variety of biological activities of ginsan have been studied, the effects of ginsan on spleen cells are not fully elucidated. We investigated the effect of ginsan on the viability and proliferation of spleen cells. Using Cell Counting Kit-8(R) solution and trypan blue solution, we found that ginsan significantly enhanced viability and proliferation. Multiple clusters, indicating proliferation, were observed in ginsan-treated spleen cells and, carboxyfluorescein succinimidyl ester and surface marker staining assay revealed that ginsan promoted proliferation from CD19+ B cells rather than CD4+ or CD8+ T cells. In addition, ginsan decreased the percentage of late apoptotic cells. Ginsan increased the surface expression of CD25 and CD69 as well as production of interleukin-2 from spleen cells, suggesting increased activation. Taken together, these results demonstrate that ginsan increases the viability and proliferation of spleen cells via multiple mechanisms, valuable information for broadening the use of ginsan in clinical and research settings.
Animals ; B-Lymphocytes ; Cell Count ; Diminazene ; Fluoresceins ; Interleukin-2 ; Mice ; Panax ; Polysaccharides ; Spleen ; T-Lymphocytes ; Trypan Blue

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Modulation of Presynaptic GABA Release by Oxidative Stress in Mechanically-isolated Rat Cerebral Cortical Neurons.

Eu Teum HAHM ; Jung Woo SEO ; Jinyoung HUR ; Young Wuk CHO

The Korean Journal of Physiology and Pharmacology.2010;14(3):127-132. doi:10.4196/kjpp.2010.14.3.127

Reactive oxygen species (ROS), which include hydrogen peroxide (H2O2), the superoxide anion (O2-.), and the hydroxyl radical (OH.), are generated as by-products of oxidative metabolism in cells. The cerebral cortex has been found to be particularly vulnerable to production of ROS associated with conditions such as ischemia-reperfusion, Parkinson's disease, and aging. To investigate the effect of ROS on inhibitory GABAergic synaptic transmission, we examined the electrophysiological mechanisms of the modulatory effect of H2O2 on GABAergic miniature inhibitory postsynaptic current (mIPSCs) in mechanically isolated rat cerebral cortical neurons retaining intact synaptic boutons. The membrane potential was voltage-clamped at -60 mV and mIPSCs were recorded and analyzed. Superfusion of 1-mM H2O2 gradually potentiated mIPSCs. This potentiating effect of H2O2 was blocked by the pretreatment with either 10,000-unit/mL catalase or 300-micrometer N-acetyl-cysteine. The potentiating effect of H2O2 was occluded by an adenylate cyclase activator, forskolin, and was blocked by a protein kinase A inhibitor, N-(2-[p-bromocinnamylamino] ethyl)-5-isoquinolinesulfonamide hydrochloride. This study indicates that oxidative stress may potentiate presynaptic GABA release through the mechanism of cAMP-dependent protein kinase A (PKA)-dependent pathways, which may result in the inhibition of the cerebral cortex neuronal activity.
Adenylyl Cyclases ; Aging ; Animals ; Catalase ; Cerebral Cortex ; Cyclic AMP-Dependent Protein Kinases ; Forskolin ; gamma-Aminobutyric Acid ; Hydrogen Peroxide ; Hydroxyl Radical ; Inhibitory Postsynaptic Potentials ; Membrane Potentials ; Neurons ; Oxidative Stress ; Parkinson Disease ; Presynaptic Terminals ; Rats ; Reactive Oxygen Species ; Superoxides ; Synaptic Transmission

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A Carbohydrate Fraction, AIP1, from Artemisia Iwayomogi Reduces the Action Potential Duration by Activation of Rapidly Activating Delayed Rectifier K+ Channels in Rabbit Ventricular Myocytes.

Won Sun PARK ; Youn Kyoung SON ; Eun A KO ; Seong Woo CHOI ; Nari KIM ; Tae Hoon CHOI ; Hyun Joo YOUN ; Su Hyun JO ; Da Hye HONG ; Jin HAN

The Korean Journal of Physiology and Pharmacology.2010;14(3):119-125. doi:10.4196/kjpp.2010.14.3.119

We investigated the effects of a hot-water extract of Artemisia iwayomogi, a plant belonging to family Compositae, on cardiac ventricular delayed rectifier K+ current (I(K)) using the patch clamp technique. The carbohydrate fraction AIP1 dose-dependently increased the heart rate with an apparent EC(50) value of 56.1+/-5.5 microgram/ml. Application of AIP1 reduced the action potential duration (APD) in concentration-dependent fashion by activating I(K) without significantly altering the resting membrane potential (IC(50) value of APD(50): 54.80+/-2.24, IC(50) value of APD(90): 57.45+/-3.47 microgram/ml). Based on the results, all experiments were performed with 50 microgram/ml of AIP1. Pre-treatment with the rapidly activating delayed rectifier K+ current (I(Kr)) inhibitor, E-4031 prolonged APD. However, additional application of AIP1 did not reduce APD. The inhibition of slowly activating delayed rectifier K+ current (I(Ks)) by chromanol 293B did not change the effect of AIP1. AIP1 did not significantly affect coronary arterial tone or ion channels, even at the highest concentration of AIP1. In summary, AIP1 reduces APD by activating I(Kr) but not I(Ks). These results suggest that the natural product AIP1 may provide an adjunctive therapy of long QT syndrome.
Action Potentials ; Artemisia ; Asteraceae ; Chromans ; Diphosphonates ; Heart Rate ; Humans ; Ion Channels ; Long QT Syndrome ; Membrane Potentials ; Muscle Cells ; Piperidines ; Plants ; Pyridines ; Sulfonamides

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Effects of Saccharin Intake on Hippocampal and Cortical Plasticity in Juvenile and Adolescent Rats.

Jong Sil PARK ; Sang Bae YOO ; Jin Young KIM ; Sung Joong LEE ; Seog Bae OH ; Joong Soo KIM ; Jong Ho LEE ; Kyungpyo PARK ; Jeong Won JAHNG ; Se Young CHOI

The Korean Journal of Physiology and Pharmacology.2010;14(2):113-118. doi:10.4196/kjpp.2010.14.2.113

The sensory system is developed and optimized by experiences given in the early phase of life in association with other regions of the nervous system. To date, many studies have revealed that deprivation of specific sensory experiences can modify the structure and function of the central nervous system; however, the effects of sensory overload remains unclear. Here we studied the effect of overloading the taste sense in the early period of life on the synaptic plasticity of rat hippocampus and somatosensory cortex. We prepared male and female Sprague Dawley rats with ad libitum access to a 0.1% saccharin solution for 2 hrs per day for three weeks after weaning on postnatal day 22. Saccharin consumption was slightly increased in males compared with females; however, saccharin intake did not affect chow intake or weight gain either in male or in female rats. We examined the effect of saccharin-intake on long term potentiation (LTP) formation in hippocampal Schaffer collateral pathway and somatosensory cortex layer IV - II/III pathways in the 6-week old saccharin-fed rats. There was no significant difference in LTP formation in the hippocampus between the control group and saccharin-treated group in both male and female rats. Also in the somatosensory cortex, we did not see a significant difference in LTP among the groups. Therefore, we conclude that saccharin-intake during 3~6 weeks may not affect the development of physiological function of the cortical and hippocampal synapses in rats.
Adolescent ; Animals ; Female ; Hippocampus ; Humans ; Long-Term Potentiation ; Male ; Nervous System ; Plastics ; Rats ; Rats, Sprague-Dawley ; Saccharin ; Somatosensory Cortex ; Synapses ; Weaning ; Weight Gain

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Caffeine and 2-Aminoethoxydiphenyl Borate (2-APB) Have Different Ability to Inhibit Intracellular Calcium Mobilization in Pancreatic Acinar Cell.

Kyung Jin CHOI ; Kab Sung KIM ; Se Hoon KIM ; Dong Kwan KIM ; Hyung Seo PARK

The Korean Journal of Physiology and Pharmacology.2010;14(2):105-111. doi:10.4196/kjpp.2010.14.2.105

Inositol 1,4,5-trisphosphate receptors (InsP3Rs) modulate Ca2+ release from intracellular Ca2+ store and are extensively expressed in the membrane of endoplasmic/sarcoplasmic reticulum and Golgi. Although caffeine and 2-aminoethoxydiphenyl borate (2-APB) have been widely used to block InsP3Rs, the use of these is limited due to their multiple actions. In the present study, we examined and compared the ability of caffeine and 2-APB as a blocker of Ca2+ release from intracellular Ca2+ stores and Ca2+ entry through store-operated Ca2+ (SOC) channel in the mouse pancreatic acinar cell. Caffeine did not block the Ca2+ entry, but significantly inhibited carbamylcholine (CCh)-induced Ca2+ release. In contrast, 2-APB did not block CCh-induced Ca2+ release, but remarkably blocked SOC-mediated Ca2+ entry at lower concentrations. In permeabilized acinar cell, caffeine had an inhibitory effect on InsP3-induced Ca2+ release, but 2-APB at lower concentration, which effectively blocked Ca2+ entry, had no inhibitory action. At higher concentrations, 2-APB has multiple paradoxical effects including inhibition of InsP3-induced Ca2+ release and direct stimulation of Ca2+ release. Based on the results, we concluded that caffeine is useful as an inhibitor of InsP3R, and 2-APB at lower concentration is considered a blocker of Ca2+ entry through SOC channels in the pancreatic acinar cell.
Acinar Cells ; Animals ; Boron Compounds ; Caffeine ; Calcium ; Carbachol ; Inositol 1,4,5-Trisphosphate Receptors ; Membranes ; Mice ; Reticulum

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Differential Expression of Metabolism-related Genes in Liver of Diabetic Obese Rats.

Eunhui SEO ; Eun Jin PARK ; Mi Kyoung PARK ; Duk Kyu KIM ; Hye Jeong LEE ; Sook Hee HONG

The Korean Journal of Physiology and Pharmacology.2010;14(2):99-103. doi:10.4196/kjpp.2010.14.2.99

The Otsuka Long-Evans Tokushima Fatty (OLETF) rat, a model of spontaneous type 2 diabetes (T2D), develops hyperglycemic obesity with hyperinsulinemia and insulin resistance after the age of 25 weeks, similar to patients with noninsulin-dependent diabetes mellitus (DM). In the present study, we determined whether there are differences in the pattern of gene expression related to glucose and lipid metabolism between OLETF rats and their control counterparts, Long-Evans Tokushima (LETO) rats. The experiment was done using 35-week-old OLETF and LETO rats. At week 35 male OLETF rats showed overt T2D and increases in blood glucose, plasma insulin, plasma triglycerides (TG) and plasma total cholesterol (TC). Livers of diabetic OLETF and LETO rats also showed differences in expression of mRNA for glucose and lipid metabolism related genes. Among glucose metabolism related genes, GAPDH mRNA was significantly higher and FBPase and G6Pase mRNA were significantly lower in OLETF rats. For lipid metabolism related genes, HMGCR, SCD1 and HL mRNA were substantially higher in OLETF rats. These results indicate that gluconeogenesis in OLETF rats is lower and glycolysis is higher, which means that glucose metabolism might be compensated for by a lowering of the blood glucose level. However, lipid synthesis is increased in OLETF rats so diabetes may be aggravated. These differences between OLETF and LETO rats suggest mechanisms that could be targeted during the development of therapeutic agents for diabetes.
Animals ; Blood Glucose ; Cholesterol ; Diabetes Mellitus, Type 2 ; Gene Expression ; Gluconeogenesis ; Glucose ; Glycolysis ; Humans ; Hyperinsulinism ; Insulin ; Insulin Resistance ; Lipid Metabolism ; Liver ; Male ; Obesity ; Plasma ; Rats ; Rats, Inbred OLETF ; RNA, Messenger ; Triglycerides

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Pyrithione-zinc Prevents UVB-induced Epidermal Hyperplasia by Inducing HIF-1alpha.

Young Suk CHO ; Kyung Hoon LEE ; Jong Wan PARK

The Korean Journal of Physiology and Pharmacology.2010;14(2):91-97. doi:10.4196/kjpp.2010.14.2.91

Epidermal keratinocytes overgrow in response to ultraviolet-B (UVB), which may be associated with skin photoaging and cancer development. Recently, we found that HIF-1alpha controls the keratinocyte cell cycle and thereby contributes to epidermal homeostasis. A further study demonstrated that HIF-1alpha is down-regulated by UVB and that this process is involved in UVB-induced skin hyperplasia. Therefore, we hypothesized that the forced expression of HIF-1alpha in keratinocytes would prevent UVB-induced keratinocyte overgrowth. Among several agents known to induce HIF-1alpha, pyrithione-zinc (Py-Zn) overcame the UVB suppression of HIF-1alpha in cultured keratinocytes. Mechanistically, Py-Zn blocked the degradation of HIF-1alpha protein in keratinocytes, while it did not affect the synthesis of HIF-1alpha. Moreover, the p21 cell cycle inhibitor was down-regulated after UVB exposure, but was robustly induced by Py-Zn. In mice repeatedly irradiated with UVB, the epidermis became hyperplastic and HIF-1alpha disappeared from nuclei of epidermal keratinocytes. However, a cream containing Py-Zn effectively prevented the skin thickening and up-regulated HIF-1alpha to the normal level. These results suggest that Py-Zn is a potential agent to prevent UVB-induced photoaging and skin cancer development. This work also provides insight into a molecular target for treatment of UVB-induced skin diseases.
Animals ; Cell Cycle ; Epidermis ; Homeostasis ; Hyperplasia ; Keratinocytes ; Mice ; Skin ; Skin Diseases ; Skin Neoplasms

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The Inhibitory Effects of Hydrogen Sulfide on Pacemaker Activity of Interstitial Cells of Cajal from Mouse Small Intestine.

Shankar Prasad PARAJULI ; Seok CHOI ; Jun LEE ; Young Dae KIM ; Chan Guk PARK ; Man Yoo KIM ; Hyun Il KIM ; Cheol Ho YEUM ; Jae Yeoul JUN

The Korean Journal of Physiology and Pharmacology.2010;14(2):83-89. doi:10.4196/kjpp.2010.14.2.83

In this study, we studied whether hydrogen sulfide (H2S) has an effect on the pacemaker activity of interstitial cells of Cajal (ICC), in the small intestine of mice. The actions of H2S on pacemaker activity were investigated using whole-cell patch-clamp technique, intracellular Ca2+ analysis at 30degrees C and RT-PCR in cultured mouse intestinal ICC. Exogenously applied sodium hydrogen sulfide (NaHS), a donor of hydrogen sulfide, caused a slight tonic inward current on pacemaker activity in ICC at low concentrations (50 and 100 micrometer), but at high concentration (500 micrometer and 1 mM) it seemed to cause light tonic inward currents and then inhibited pacemaker amplitude and pacemaker frequency, and also an increase in the resting currents in the outward direction. Glibenclamide or other potassium channel blockers (TEA, BaCl2, apamin or 4-aminopydirine) did not have an effect on NaHS-induced action in ICC. The exogenous application of carbonilcyanide p-triflouromethoxyphenylhydrazone (FCCP) and thapsigargin also inhibited the pacemaker activity of ICC as NaHS. Also, we found NaHS inhibited the spontaneous intracellular Ca2+ ([Ca2+]i) oscillations in cultured ICC. In doing an RT-PCR experiment, we found that ICC enriched population lacked mRNA for both CSE and CBS, but was prominently detected in unsorted muscle. In conclusion, H2S inhibited the pacemaker activity of ICC by modulating intracellular Ca2+. These results can serve as evidence of the physiological action of H2S as acting on the ICC in gastrointestinal (GI) motility.
Animals ; Apamin ; Barium Compounds ; Chlorides ; Gastrointestinal Motility ; Glyburide ; Humans ; Hydrogen ; Hydrogen Sulfide ; Interstitial Cells of Cajal ; Intestine, Small ; Light ; Mice ; Muscles ; Patch-Clamp Techniques ; Potassium Channel Blockers ; RNA, Messenger ; Sodium ; Sulfides ; Thapsigargin ; Tissue Donors

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Fenofibrate Reduces Age-related Hypercholesterolemia in Normal Rats on a Standard Diet.

Ying HAN ; Mi Hyang DO ; Mi Sun KIM ; Eunhui SEO ; Mi Kyoung PARK ; Duk Kyu KIM ; Hye Jeong LEE ; Su Yeong SEO

The Korean Journal of Physiology and Pharmacology.2010;14(2):77-81. doi:10.4196/kjpp.2010.14.2.77

Plasma cholesterol is increased in normal aging in both rodents and humans. This is associated with reduced elimination of cholesterol and decreased receptor-mediated clearance of plasma low-density lipoprotein (LDL) cholesterol. The aims of this study were: (1) to determine age-related changes in plasma lipid profiles, and (2) to determine the effect of fenofibrate, an activator of peroxisome proliferator activated receptor alpha (PPAR alpha), on plasma lipid profiles in normal rats on a standard diet. Male Sprague-Dawley (SD) rats (n=15) were fed standard chow and water from 10 to 25 weeks of age. During that period, we measured daily food intake, body weight, fasting and random blood glucose levels, plasma total cholesterol (TC), triglycerides (TG), and free fatty acid (FFA) levels. At 20 weeks of age, all rats were randomly divided into two groups: a fenofibrate group (in which rats were gavaged with 300 mg/kg/day of fenofibrate) and a control group (gavaged with water). Fenofibrate treatment lasted 5 weeks. There were no significant changes in daily food intake, blood glucose, and plasma TG level with age. Body weight, plasma TC, and FFA levels were significantly increased with age. Fenofibrate significantly decreased plasma concentrations of TC and FFA, which had been increased with age. However, fenofibrate did not influence the plasma concentration of TG, which had not increased with age. These results suggest that fenofibrate might have a novel role in preventing age-related hypercholesterolemia in SD rats on a normal diet.
Aging ; Animals ; Blood Glucose ; Body Weight ; Cholesterol ; Diet ; Eating ; Fasting ; Fenofibrate ; Humans ; Hypercholesterolemia ; Lipoproteins ; Male ; Plasma ; PPAR alpha ; Rats ; Rats, Sprague-Dawley ; Rodentia ; Triglycerides ; Water

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Intestinal Absorption of Fibrinolytic and Proteolytic Lumbrokinase Extracted from Earthworm, Eisenia andrei.

Xiang Mei YAN ; Chung Hyo KIM ; Chul Kyu LEE ; Jang Sik SHIN ; Il Hwan CHO ; Uy Dong SOHN

The Korean Journal of Physiology and Pharmacology.2010;14(2):71-75. doi:10.4196/kjpp.2010.14.2.71

To investigate the intestinal absorption of a fibrinolytic and proteolytic lumbrokinase extracted from Eisenia andrei, we used rat everted gut sacs and an in situ closed-loop recirculation method. We extracted lumbrokinase from Eisenia andrei, and then raised polyclonal antibody against lumbrokinase. Fibrinolytic activity and proteolytic activity in the serosal side of rat everted gut sacs incubated with lumbrokinase showed dose- and time-dependent patterns. Immunological results obtained by western blotting serosal side solution using rat everted gut sacs method showed that lumbrokinase proteins between 33.6 and 54.7 kDa are absorbed mostly by the intestinal epithelium. Furthermore, MALDI-TOF mass spectrometric analysis of plasma fractions obtained by in situ recirculation method confirmed that lumbrokinase F1 is absorbed into blood. These results support the notion that lumbrokinase can be absorbed from mucosal lumen into blood by oral administration.
Administration, Oral ; Animals ; Blotting, Western ; Endopeptidases ; Intestinal Absorption ; Intestinal Mucosa ; Oligochaeta ; Plasma ; Proteins ; Rats

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