Hyoung Jin Park

Afferent innervation of the rat pancreas : Retrograde tracing and immunohistochemistry in the dorsal root ganglia

This study was undertaken to determine the segmental organization of the dorsal root ganglion (DRG) cells that give rise to pancreatic afferents containing a certain neuropeptide in the rat. These cells were examined using retrograde tracing combined with immunohistochemistry. Injection of horseradish peroxidase (HRP) into the pancreas resulted in the labeling of cells in bilateral T5-L2 DRGs, with most labeled cells lying at T10-T11. Injection into the duodenal (right), splenic (left), and entire lobes consistently produced more labeled cells significantly in the right, left, and right DRGs, respectively. Calcitonin gene-related peptide (CGRP)-, substance P (SP)-, somatostatin (SOM)-, and galanin (GAL)-immunoreactive (IR) cells in the DRGs (T9-T12) were found in -52, 17, 8, and 6%, respectively, but neuropeptide Y- and vasoactive intestinal polypeptide-IR cells were not found. About 88% of HRP-labeled cells in DRGs (T9-T12) contained CGRP, and approximately 16% of them contained SP. Although SOM- and GAL-IR cells were localized in the DRGs, these cells innervating the pancreas could not be found. In brief, these results show that bilateral (not similar in cell number on each side) DRG cells innervate the duodenal or splenic pancreas, and the majority of these cells that project to the pancreas contain CGRP and SP.

Significant cholinergic role in secretin-stimulated exocrine secretion in isolated rat pancreas

Effects of intrapancreatic cholinergic activation by electrical field stimulation (EFS) on secretin-stimulated pancreatic exocrine secretion were investigated in the totally isolated perfused rat pancreas. EFS at 15 V, 2 ms, and 8 Hz for 45 min markedly increased spontaneous pancreatic secretion. This increase was completely inhibited by tetrodotoxin (1 μM) but not by hexamethonium (100 μM). Atropine (2 μM) significantly reduced the EFS-evoked volume flow and amylase output by 52% and 80%, respectively. EFS further increased the secretin (12 pM)-stimulated pancreatic secretion of fluid and amylase. The increases of the two parameters were significantly suppressed by atropine by 28% and 72%, respectively. Interestingly, EFS significantly increased concentrations of somatostatin-like immunoreactivity in portal venous effluents. When pertussis toxin (200 ng/ml) or rabbit antisomatostatin serum (0.1 ml/10 ml; titer of 1:50,000) was intra-arterially administered, EFS further increased the secretin-stimulated pancreatic secretion. In conclusion, the activation of intrapancreatic cholinergic neurons potentiated the secretin action on pancreatic exocrine secretion in the rat. This potentiating effect was significantly reduced by local somatostatin released during EFS that activated intrapancreatic cholinergic tone.

Inhibition of 2,4-dinitrofluorobenzene-induced atopic dermatitis by topical application of the butanol extract of Cordyceps bassiana in NC/Nga mice

ETHNOPHARMACOLOGICAL SIGNIFICANCE The Cordyceps species are insect-borne mushrooms that have been ethnopharmacologically used for skin diseases such as eczema and dermatitis. AIM OF THE STUDY In this study, we investigated the curative effects of the butanol fraction (CBBF) of Cordyceps bassiana on atopic dermatitis. MATERIALS AND METHODS Dermatitis was induced by repeated application of 2,4-dinitrofluorobenzene (DNFB) in NC/Nga mice. After a topical application of CBBF on the skin lesions, the dermatitis score, epidermal thickness, mast cell number, and interleukin (IL)-4 and interferon (IFN)-γ, as well as the levels of histamine and immunoglobulin E (IgE) in the serum, were measured. Moreover, effect of CBBF on histamine release was examined using RBL-2H3 under stimulation with 2,4-dinitrophenylated bovine serum albumin (DNP-BSA). RESULTS CBBF inhibited atopic dermatitis symptoms and signs in the DNFB-treated NC/Nga mice. The suppressive activity of topically applied CBBF may be due to the dose-dependent blockade of a series of immunopathological events, including the release of histamine, the production of IgE, and the secretion of IL-4 and IFN-γ. However, this extract did not directly suppress the degranulation process, assessed by measuring β-hexosaminidase release. CONCLUSIONS Our results suggest that CBBF can be applied as an effective herbal remedy to treat atopic dermatitis.

Protective effect of Phellinus linteus polysaccharide extracts against thioacetamide-induced liver fibrosis in rats: a proteomics analysis

BackgroundThe hepatoprotective potential of Phellinus linteus polysaccharide (PLP) extracts has been described. However, the molecular mechanism of PLP for the inhibition of liver fibrosis is unclear. This study aims to investigate the molecular protein signatures involved in the hepatoprotective mechanisms of PLP via a proteomics approach using a thioacetamide (TAA)-induced liver fibrosis rat model.MethodsMale Sprague–Dawley rats were divided into three groups of six as follows: Normal group; TAA group, in which rats received TAA only; and PLP group, in which rats received PLP and TAA. Liver fibrosis was induced in the rats by repeated intraperitoneal injections of TAA at a dose of 200 mg/kg body weight twice a week for 4 weeks. PLP was given orally at a dose of 50 mg/kg body weight twice a day from the beginning of the TAA treatment until the end of the experiment. The development of liver cirrhosis was verified by histological examination. Liver proteomes were established by two-dimensional gel electrophoresis. Proteins with significantly altered expression levels were identified by matrix-assisted laser desorption/ionization-time of flight/time of flight mass spectrometry and the differentially expressed proteins were validated by immunohistochemical staining and reverse transcription polymerase chain reaction.ResultsHistological staining showed a remarkable reduction in liver fibrosis in the rats with PLP treatment. A total of 13 differentially expressed proteins including actin, tubulin alpha-1C chain, preprohaptoglobin, hemopexin, galectin-5, glutathione S-transferase alpha-4 (GSTA4), branched chain keto acid dehydrogenase hterotetrameric E1 subunit alpha (BCKDHA), glutathione S-transferase mu (GSTmu); glyceraldehyde-3-phosphate dehydrogenase (GAPDH); thiosulfate sulfurtransferase (TFT); betaine-homocysteine S-methyltransferase 1 (BHMT1); quinoid dihydropteridine reductase (QDPR); ribonuclease UK114 were observed between the TAA and PLP groups. These proteins are involved in oxidative stress, heme and iron metabolism, cysteine metabolism, and branched-chain amino acid catabolism.ConclusionThe proteomics data indicate that P. linteus may be protective against TAA-induced liver fibrosis via regulation of oxidative stress pathways, heat shock pathways, and metabolic pathways for amino acids and nucleic acids.

Effects of pancreatic polypeptide on insulin action in exocrine secretion of isolated rat pancreas.

1. Effects of pancreatic polypeptide (PP) on insulin action in pancreatic exocrine secretion was investigated by using an isolated rat pancreas that was perfused with Krebs‐Henseleit solution containing 2.5 mM glucose, 0.1% bovine serum albumin and 3% Dextran T‐70 at a vascular flow rate of 1.2 ml min‐1. 2. Cholecystokinin‐8 (CCK‐8) at a concentration of 14 pM stimulated basal flow rate and amylase output of the isolated pancreas. Twenty‐five millimolar glucose not only increased the basal flow rate and amylase output but also potentiated the CCK‐stimulated flow rate and amylase output. 3. Porcine insulin, administered intra‐arterially at a concentration of 100 nM, also increased the basal flow rate and amylase output, and also potentiated the CCK‐stimulated flow rate and amylase output. 4. Rat PP, given intra‐arterially at a concentration of 10 pM, completely abolished the potentiation effects of both the 25 mM glucose and the exogenous insulin on the CCK‐stimulated flow rate and amylase output. Rat PP also inhibited the flow rate and amylase output increased by either 25 mM glucose alone or exogenous insulin alone. However, rat PP did not change the flow rate and amylase output stimulated by CCK‐8 alone. 5. These results indicate that insulin is an important stimulatory hormone of pancreatic exocrine secretion, and that PP exerts the inhibitory role in pancreatic exocrine secretion by modulating the insulin action.

p38-Targeted inhibition of interleukin-12 expression by ethanol extract from Cordyceps bassiana in lipopolysaccharide-activated macrophages

Cordyceps species have been known as ethnopharmacologically valuable mushroom in Korea, China, and Japan. This plant has been reported to exhibit a variety of pharmacological activities such as antioxidative, anticancer, anti-inflammatory, antidiabetic, and antiobesity effects. Although numerous pharmacological potentials of Cordyceps spp. have been demonstrated, immunomodulatory effect of Cordyceps bassiana has not been published yet. To evaluate its immunomodulatory activity, macrophages activated by lipopolysaccharide (LPS) were employed and the production of interleukin-12 (IL-12) was explored in terms of understanding its molecular inhibitory mechanism. Seventy percent of ethanol extract from Cordyceps bassiana (Cb-EE) was able to suppress the expression of IL-12, a cytokine regulating interferon-γ (IFN-γ)-producing T helper type 1 (Th1) polarization response, at the transcriptional levels. The inhibitory effect of Cb-EE seemed to be due to activator protein-1 (AP-1) translocation inhibition, according to immunoblotting analysis with nuclear fraction and luciferase assay. In agreement with this, Cb-EE strongly suppressed the phosphorylation of p38, a prime signal to stimulate AP-1 translocation and IL-12 production, strongly suppressed by SB203580, a p38 inhibitor. Furthermore, this extract also suppressed IFN-γ production in both phytohemaglutinin A and LPS-activated splenocytes. Our results suggest that Cb-EE can be applied as a Th1 response regulatory herbal medicine.

The role of insulin in the interaction of secretin and cholecystokinin in exocrine secretion of the isolated perfused rat pancreas

To investigate the role of insulin in the potentiation effect of secretin and cholecystokinin (CCK) on pancreatic exocrine secretion, the pancreas was isolated from rats and perfused with modified Krebs-Henseleit solution containing glucose at three concentrations. Intraarterial glucose at concentrations of 2.5, 10, and 25 mM produced modest but significant increases in both the pancreatic flow rate and the amylase output in a concentration-dependent manner. The mixture of secretin and CCK at concentrations of 18.5 and 14 pM, respectively, added to the glucose solutions augmented the pancreatic flow rate and amylase output in relation to the glucose concentration. In the streptozotocin-treated pancreas, the mixture of secretin and CCK failed to augment the pancreatic exocrine secretion unless exogenous insulin was added to the perfusate. Secretin markedly potentiated the CCK-induced amylase output when insulin was present in the circulation. However, CCK did not potentiate the secretin-induced flow rate even if insulin was present in the circulation. Insulin did not affect the actions of secretin alone but it potentiated the actions of CCK alone in both the pancreatic flow rate and the amylase output. It is concluded from the above results that insulin intensifies the combined actions of secretin and CCK in pancreatic exocrine secretion by potentiating the CCK action. Furthermore, in the presence of insulin, secretin is able to potentiate the pancreatic enzyme secretion stimulated by CCK.

The Anti-influenza Virus Effect of Phellinus igniarius Extract

Herbal medicine has been used in the orient for thousands of years to treat large and small ailments, including microbial infections. Although there are treatments for influenza virus infection, there is no treatment for drug-resistant viruses. It is time that we explored and exploited the multi-component nature of herbal extracts as multi-drug combination therapies. Here, we present data on the anti-influenza virus effect of a medicinal mushroom, Phellinus igniarius. The P. igniarius water extract was effective against influenza A and B viruses, including 2009 pandemic H1N1, human H3N2, avian H9N2, and oseltamivir-resistant H1N1 viruses. Virological assays revealed that the extract may interfere with one or more early events in the influenza virus replication cycle, including viral attachment to the target cell. Therefore, our results provide new insights into the use of P. igniarius as an anti-influenza medicine.

Effects of intrapancreatic neuronal activation on cholecystokinin-induced exocrine secretion of isolated perfused rat pancreas

Abstract The role of intrapancreatic neurons in the action of cholecystokinin (CCK) on pancreatic exocrine secretion of the totally isolated, perfused rat pancreas was investigated. Intrapancreatic neurons were activated by applying electrical field stimulation (EFS) to the isolated pancreas for 45 min. When applying EFS, spontaneous pancreatic secretions of fluid and amylase increased until the second 15-min period of EFS and then decreased during the third 15-min period. Atropine (2 µM) notably reduced the EFS-evoked pancreatic secretions of fluid and amylase. The CCK-induced (10 pM) pancreatic secretions of fluid and amylase elevated further in the first 15-min period of EFS and then gradually resumed to the levels observed during application of CCK alone in the third 15-min period of EFS. However, the CCK-induced pancreatic secretions remained elevated even in the third 15-min period of EFS when an action of endogenous somatostatin was inhibited by cyclo-(7-aminoheptanonyl-Phe-d-Trp-Lys-Thr[BZL]) (10 nM) or pertussis toxin (200 ng/ml). EFS further elevated spontaneous exocrine secretion by the cysteamine-treated (300 mg/kg) pancreas, but this was markedly reduced, to normal levels, by infusing somatostatin (100 pM). EFS increased the numbers of immunoreactive somatostatin cells in the Langerhans’ islets. The results indicate that intrapancreatic neuronal activation influences CCK-induced pancreatic secretions in a dual-phase pattern in the rat: an increase during the early phase and a decrease during the late phase. Endogenous somatostatin released from the islets appears to inhibit the enhancing effect of neuronal activation on CCK-induced pancreatic secretion. Of the intrapancreatic neurons, the cholinergic ones appear to predominate in EFS’s effects on CCK-induced pancreatic secretion.

Potentiation by capsaicin of lidocaine's tonic impulse block in isolated rat sciatic nerve

Compound action potentials (CAPs) of A- and C-fibers were recorded from isolated sciatic nerves of the rat to determine whether tonic block of impulse conduction induced by lidocaine was affected by low doses of capsaicin. Capsaicin alone (50 microM) did not change the CAPs of either A- or C-fibers. Although the lower concentrations of capsaicin (5-30 microM) caused no change of the tonic blocking action of lidocaine, 30 min of 50 microM capsaicin administration did induce a significant potentiation of tonic block. Capsaicins potentiating effects were partially reversed after 30 min of wash. These results suggest that capsaicin may be a useful agent for the potentiation of impulse blockade by lidocaine.