• The Korean Journal of Physiology and Pharmacology
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• 제8권5호
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• pp.237-243
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• 2004

Glial cells are activated in neuropathy and play a key role in hyperalgesia and allodynia. This study was performed to determine whether minocycline could attenuate heat hyperalgesia and mechanical allodynia, and how glial cell activation and nuclear factor kappa B (NF-kappaB) were regulated by minocycline in a model of chronic constriction of sciatic nerve (CCl). When minocycline (50 mg/kg, oral) was daily administered from 1 day before to 9 days after ligation, heat hyperalgesia and mechanical allodynia were attenuated. Furthermore, when minocycline treatment was initiated 1 or 3 days after ligation, attenuation of the hypersensitive behavior was still robust. However, the effect of attenuation was less when minocycline was started from day 5. In order to elucidate the mechanism of pain attenuation by minocycline, we examined the changes of glia and NF-kappaB, and found that attenuated hyperalgesia and allodynia by minocycline was accompanied by reduced microglial activation. Furthermore, the number of NF-kappaB immunoreactive cells increased after CCI treatment and this increase was attenuated by minocycline. We also observed translocation of NF-kappaB into the nuclei of activated glial cells. These results suggest that minocycline inhibits activation of glial cells and NF-kappaB, thereby attenuating the development of behavioral hypersensitivity to stimuli.

• Journal of Korean Neurosurgical Society
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• 제42권6호
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• pp.462-469
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• 2007

Objective : This study characterized the neurons in the lumbosacral cord that express phospho ERK (pERK) after distension or irritation of the bladder, and their relation to the vanilloid receptor 1 (VR1) positive primary afferents. Methods : Mechanical distension and chemical irritation of the bladder were induced by intravesical injection of the saline and mustard oil, respectively. Spinal neurons expressing pERK and the primary afferent fibers were characterized using multiple immunofluorescence for neurokinin 1 (NK1), neuronal nitric oxide synthetase (nNOS) and VR1. Results : Neurons in lamina I, medial dorsal horn (MDH), dorsal gray commissure (DGC) and sacral parasympathetic nucleus (SPN) were immunoreactive for pERK after either mechanical or chemical stimulation. The majority of pERK positive cells were positive for NK1 in lamina I and SPN, but not in the DGC. Most of pERK positive cells are not stained for nNOS except in a small population of the cells in the SPN and DGC. Contacts between perikarya and dendrites of pERK-positive cells and terminals of primary afferents expressing VR1 were identified in lamina I. lateral collateral path (LCP) and SPN. Conclusion : In this study, the lumbosacral neurons activated by mechanical and chemical stimulation of the urinary bladder were identified with expression of the pERK, and also provided the evidence that VR1-positive primary afferents may mediate the activation of these neurons.

• 대한수의학회지
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• 제41권2호
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• pp.123-132
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• 2001

ICR 마우스 위장관 8개 부위(위저부, 유문부, 십이지장, 공장, 회장, 맹장, 결장 및 직장)에서 위장관내분비 세포의 부위별 분포 및 상대적 빈도를 somatostain, serotonin, glucagon, chloecystokinin(CCK)-8, secretin, pancreatic polypeptide(PP) 및 gastrin 등 총 7종류의 항혈청을 이용한 면역조직화학적 방법으로 관찰하였던 결과 somatostain, serotonin, glucagon, CCK-8, secretin, 및 gastrin 면역반응세포의 7종류의 내분비세포가 관찰되었다. 본 실험의 결과 장관부위에서는 주로 타원형 또는 방추형의 개방형 세포(open-typed cell)들이 관찰된 반면 위저부와 유문부에서는 주로 원형의 폐쇄형 세포(close-typed cell)을이 관찰되었다. 이들 면역반응세포들의 부위별 분포는 위장관 각 부위에 따라 매우 다양하게 관찰되었다. Somatostain 면역반응세포들은 대장을 제외한 위장관에서 전 부위에서 관찰되었고, serotonin 면역반응세포들은 전 위장관에 걸쳐 관찰되었으며, ICR 마우스에서 가장 높은 빈도를 나타내었다. Glucagon 면역반응세포들은 위저부와 직장에 국한되어 관찰되었으며, 각각 중등도 및 소수의 빈도를 나타내었다. CCK-8 면역반응세포들은 유문부, 십이지장 및 회장에서 각각 다수, 중등도 및 극소수의 빈도로 관찰되었다. 한편 secretin 면역반응세포들은 각각 소수 및 극소수의 빈도로 십이지장과 회장에 국한되어 출현하였고, gastrin 면역반응세포들은 유문부에 국한되어 다수 관찰되었다. 그러나 PP 면역반응세포들은 전 위장관에 걸쳐 관찰되지 않았다. 결론적으로 ICR 마우스의 위장관내분비세포의 부위별 분포 및 상대적 빈도는 다른 포유동물과 유사하게 관찰되었으나, 일부 특이한 양상을 나타내기도 하였다.

• 대한의생명과학회지
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• 제16권3호
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• pp.201-206
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• 2010

In order to develop procedures for the rapid isolation of recombinant sugar transporter in functional form from away from the endogenous insect cell transporter, gene fusion techniques were exploited. Briefly, BamH1-digested human HepG2 type glucose transport protein cDNA was first cloned into a transfer vector pBlueBacHis, containing a tract of six histidine residues. Recombinant baculoviruses including the human cDNA were then generated by allelic exchange following transfection of insect cells with wild-type BaculoGold virus DNA and the recombinant transfer vector. Plaque assay was then performed to obtain and purify recombinant viruses expressing the human transport protein. All the cell samples that had been infected with viruses from the several blue plaques exhibited a positive reaction in the immnuassay, demonstrating expression of the glucose transport protein. In contrast, no color development in the immunoassay was observed for cells infected with the wild-type virus or no virus. Immunoblot analysis showed that a major immunoreactive band of apparent Mr 43,000~44,000 was evident in the lysate from cells infected with the recombinant baculovirus. Following expression of the recombinant fusion protein with the metal-binding domain and enterokinase cleavage site, the fusion protein was recovered by competition with imidizole using immobilized metal charged resin. The leader peptide was then removed from the fusion protein by cleavage with porcine enterokinase. Final separation of the recombinant protein of the interest was achieved by passage over $Ni^{2+}$-charged resin under binding conditions. The expressed transport protein bound cytochalasin B and demonstrated a functional similarity to its human counterpart.

• The Korean Journal of Physiology and Pharmacology
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• 제16권1호
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• pp.17-24
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• 2012

The hypothalamus-pituitary-adrenocortex (HPA) axis is the central mediator of the stress response. The supramammillary (SuM) region is relatively unique among the hypothalamic structures in that it sends a large, direct projection to the hippocampal formation. It has been shown that mild stress could activate the SuM cells that project to the hippocampus. However, the role of these cell populations in modulating the stress response is not known. The present study examined the effect of stress on different populations of SuM cells that project to the hippocampus by injecting the fluorescent retrograde tracer, fluorogold (FG), into the hippocampus and utilizing the immunohistochemistry of choline acetyltransferase (ChAT), corticotrophin releasing factor (CRF), serotonin (5-HT), glutamate decarboxylase (GAD), tyrosine hydroxylase (TH) and NADPH-d reactivity. Immobilization (IMO) stress (2 hr) produced an increase in the expression of ChAT- immunoreactivity, and tended to increase in CRF, 5-HT, GAD, TH-immunoreactivity and nitric oxide (NO)-reactivity in the SuM cells. Fifty-three percent of 5-HT, 31% of ChAT and 56% of CRF cells were double stained with retrograde cells from the hippocampus. By contrast, a few retrogradely labeled cells projecting to the hippocampus were immunoreactive for dopamine, ${\gamma}$-aminobutyric acid (GABA) and NO. These results suggest that the SuM region contains distinct cell populations that differentially respond to stress. In addition, the findings suggest that serotonergic, cholinergic and corticotropin releasing cells projecting to the hippocampus within the SuM nucleus may play an important role in modulating stress-related behaviors.

• The Journal of Korean Physical Therapy
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• 제27권3호
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• pp.140-146
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• 2015

Purpose: To determine the effects of 630 nm light emitting diode (LED) on full-thickness wound healing. Methods: Twelve male Sprague-Dawley rats were randomly divided into LED (n=6) and control group (n=6). Two $19.63mm^2$ wounds were created on the mid dorsum. LED group received a 630 nm LED irradiation with $3.67mW/cm^2$ for 30 minutes ($6.60J/cm^2$) for 7 days, while control group received sham LED irradiation. Epithelial gap, collagen density, ${\alpha}$-SMA fibroblast and PCNA keratinocyte were measured on histochemical and immunohistochemical staining using image analysis system. An independent t-test was conducted to compare the difference between groups. Results: The wound closure rate, collagen density, ${\alpha}$-SMA fibroblast number, epithelial gap and PCNA keratinocyte number have shown no significant difference between LED and control group at day 3 after the treatment. At day 7 after the treatment, the wound closure rate in LED group was increased when compared with control group (p<0.05). The collagen density (p<0.05) and ${\alpha}$-SMA immunoreactive fibroblast number (p<0.001) were increased when compared with control group at day 7. The epithelial gap in LED group was significantly shorten than control group at day 7 (p<0.01). The PCNA positive cell number in LED group was higher than control group at day 7 (p<0.01). Conclusion: 630 nm LED with $3.67mW/cm^2$, $6.60J/cm^2$ accelerate collagen deposition by stimulating fibroblasts, and enhance wound contraction by differentiating myofibroblasts in the dermis, and accelerate keratinocyte proliferation by facilitating DNA synthesis in the epidermis. It may promote the healing process in proliferation stage of wound healing.

• 동의생리병리학회지
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• 제18권1호
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• pp.157-166
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• 2004

In order to understand the anti-carcinogenic effects of Boo-jung-bae-bon-bang(扶正培本方)-B1), Hwal-hyul-hwa-eo-bang(活血化瘀方-B2), Cheong-youl-hae-dok-bang(淸熱解毒方-B3), prescriptions of individual B1, B2, and B3 treatments or combined treatments (B4; B1+B2, B5; B1+B3, B6; B1+B2+83, B7; B2+83) were applied to cultured cancer cell lines. The major findings on their anti-cardnogenic effects in terms of mechanism and synergistic interactions are summarized below. Results of cytokine gene expression analyses are summarized as follows; IL-2 gene expression was increased by B1 and B6 treatments, IFN-v gene expression increased by B3, B1, B6, and 85, and CD28 gene expression increased by B1 and B5. IL-4 expression was not affected by any treatments. In the FACS analysis, increases in numbers of immunoreactive CD3/sup +//CD25/sup +/ T cells by B1 and B5 treatment, CD3/sup +//CD69/sup +/ T cells by B1, B3, B5, and B6 treatments, CD19/sup +//CD44/sup +/ B cells by B1 and B6 treatments were observed when compared to the corresponding non-treated control groups.

• Molecules and Cells
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• 제19권3호
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• pp.408-417
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• 2005

Nitric oxide (NO) occurs in various types of cells in the central nervous system. We studied the distribution and morphology of neuronal nitric oxide synthase (NOS)-containing neurons in the visual cortex of mouse and rabbit with antibody immunocytochemistry. We also compared this labeling to that of calbindin D28K, calretinin, and parvalbumin. Staining for NOS was seen both in the specific layers and in selective cell types. The densest concentration of intense anti-NOS immunoreactive (IR) neurons was found in layer VI, while the weak anti-NOS-IR neurons were found in layer II/III in both animals. The NOS-IR neurons varied in morphology. The large majority of NOS-IR neurons were round or oval cells with many dendrites coursing in all directions. Two-color immunofluorescence revealed that only 16.7% of the NOS-IR cells were double-labeled with calbindin D28K in the mouse visual cortex, while more than half (51.7%) of the NOS-IR cells were double-labeled with calretinin and 25.0% of the NOS-IR cells were double-labeled with parvalbumin in mouse. By contrast, 92.4% of the NOS-IR neurons expressed calbindin D28K while only 2.5% of the NOS-IR neurons expressed calretinin in the rabbit visual cortex. In contrast with the mouse, none of the NOS-IR cells in the rabbit visual cortex were double-labeled with parvalbumin. The results indicate that neurons in the visual cortex of both animals express NOS in specific layers and cell types, which do not correlate with the expression of calbindin D28K, calretinin or parvalbumin between the two animals.

• The Korean Journal of Physiology and Pharmacology
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• 제25권4호
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• pp.365-374
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• 2021

The mammalian target of rapamycin (mTOR) plays a role in various cellular phenomena, including autophagy, cell proliferation, and differentiation. Although recent studies have reported its involvement in nociceptive responses in several pain models, whether mTOR is involved in orofacial pain processing is currently unexplored. This study determined whether rapamycin, an mTOR inhibitor, reduces nociceptive responses and the number of Fos-immunoreactive (Fos-ir) cells in the trigeminal nucleus caudalis (TNC) in a mouse orofacial formalin model. We also examined whether the glial cell expression and phosphorylated p38 (p-p38) mitogen-activated protein kinases (MAPKs) in the TNC are affected by rapamycin. Mice were intraperitoneally given rapamycin (0.1, 0.3, or 1.0 mg/kg); then, 30 min after, 5% formalin (10 μl) was subcutaneously injected into the right upper lip. The rubbing responses with the ipsilateral forepaw or hindpaw were counted for 45 min. High-dose rapamycin (1.0 mg/kg) produced significant antinociceptive effects in both the first and second phases of formalin test. The number of Fos-ir cells in the ipsilateral TNC was also reduced by high-dose rapamycin compared with vehicle-treated animals. Furthermore, the number of p-p38-ir cells the in ipsilateral TNC was significantly decreased in animals treated with high-dose rapamycin; p-p38 expression was co-localized in microglia, but not neurons and astrocytes. Therefore, the mTOR inhibitor, rapamycin, reduces orofacial nociception and Fos expression in the TNC, and its antinociceptive action on orofacial pain may be associated with the inhibition of p-p38 MAPK in the microglia.

• Asian-Australasian Journal of Animal Sciences
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• 제31권5호
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• pp.650-657
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• 2018

Objective: The study investigated the biological functions and mechanisms for controlling cashmere growth of Liaoning cashmere goat by ovarian carcinoma immunoreactive antigen-like protein 2 (OCIAD2) and decorin (DCN) genes. Methods: cDNA library of Liaoning cashmere goat was constructed in early stages. OCIAD2 and DCN genes related to cashmere growth were identified by homology analysis comparison. The expression location of OCIAD2 and DCN genes in primary and secondary hair follicles (SF) was performed using in situ hybridization. The expression of OCIAD2 and DCN genes in primary and SF was performed using real-time polymerase chain reaction (PCR). Results: In situ hybridization revealed that OCIAD2 and DCN were expressed in the inner root sheath of Liaoning cashmere goat hair follicles. Real-time quantitative PCR showed that these genes were highly expressed in SF during anagen, while these genes were highly expressed in primary hair follicle in catagen phase. Melatonin (MT) inhibited the expression of OCIAD2 and promoted the expression of DCN. Insulin-like growth factors-1 (IGF-1) inhibited the expression of OCIAD2 and DCN, while fibroblast growth factors 5 (FGF5) promoted the expression of these genes. MT and IGF-1 promoted OCIAD2 synergistically, while MT and FGF5 inhibited the genes simultaneously. MT+IGF-1/MT+FGF5 inhibited DCN gene. RNAi technology showed that OCIAD2 expression was promoted, while that of DCN was inhibited. Conclusion: Activation of bone morphogenetic protein (BMP) signaling pathway up-regulated OCIAD2 expression and stimulated SF to control cell proliferation. DCN gene affected hair follicle morphogenesis and periodic changes by promoting transforming growth $factor-{\beta}$ ($TGF-{\beta}$) and BMP signaling pathways. OCIAD2 and DCN genes have opposite effects on $TGF-{\beta}$ signaling pathway and inhibit each other to affect the hair growth.