• Title/Summary/Keyword: immunohistochemical methods

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Experimental Studies on the Hair Growth Activity of Drynariae Rhizoma Extracts in C57BL/6N Mice (C57BL/6N 생쥐에서 골쇄보(骨碎補) 추출물을 이용한 발모(發毛)효과에 대한 실험적 연구)

  • Jung, Suk-Young;Jung, Il-Kook;Kim, Dae-Keun;Jo, Han-Young;Jung, Han-Sol;Lee, Chang-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.2
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    • pp.453-461
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    • 2007
  • Drynariae Rhizoma has been used for promotes mending of the sinews and bone, tonifies the kidney for such symptoms as weak low back and knees, and stimulates the growth of hair as a tinctute for alopecia in oriental medicine. This experiment examined the effect of an acetone extracts of Drynariae Rhizomas(GSB-1), its EtoAc fraction(GB-2) and n-buOH fraction(GSB-3), on hair growth activity of the C57BL/6L mice after topical application to skin. First, We examined on hair growth activity of extracts of Drynariae Rhizomas compare to control and 1 % minoxidil groups. Second, We investigated on the number of hair follicle and mast cells after topical application of extracts of the Drynariae Rhizomas to skin for 16 day. Third, We investigated immunoreactive density of vascular endothelial growth factor(VEGF), protein kinase C-${\alpha}$(PKC-${\alpha}$) and stem(mast) cell factor(SCF) in skin of C57BL/6N mice by immunohistochemical methods. The results were as follows : Hair growth effect of acetone extracts of Drynariae Rhizomas, its EtoAc fraction and n-BuOH fraction was observed in 98 %, 96 % and 60 % in hair removed skin area in 16 day respectively, Immunoreactive density of VEGF in skin of GSB-1 group was weakly stained compare to control group in 10 day, But GSB-2 and GSB-3 groups were mildy stained in bulge and root sheath of skin. Immunolocalization of SCF antigens was observed weakly stained density in epidermis, bulge, stem cells and dermal papilla of control gruop. but in experimental group, immunoreactivity of SCF antigens was observed mildly stained density in bulge, epidermis and root sheath of GSB-1 gruop, heavily stained density in epidermis, bulge and root sheath of GSB-2 and GSB-3 groups to the hair removal skin of C57BL/6N mice on day 10. These experiment suggest that acetone extracts of Drynariae Rhizomas and its EtoAc fraction may be used for topical treatment of alopecia areata.

Anti-inflammatory Effects of Hataedock Extracted from Coptidis Rhizoma and Glycyrrhiza Uralensis on Atopic Dermatitis-like Skin Lesions of NC/Nga Mouse (황련-감초 추출물을 이용한 하태독법이 NC/Nga 생쥐에서 유발된 아토피 유사 피부염에 미치는 항염증 효과)

  • Cha, Ho-yeol;Ahn, Sang-hyun;Jeong, A-ram;Cheon, Jin-hong;Park, Sun-young;Choi, Jun-yong;Kim, Ki-bong
    • The Journal of Internal Korean Medicine
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    • v.36 no.4
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    • pp.486-497
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    • 2015
  • Objectives Hataedock is an orally administered herbal extract treatment for newborn babies that dispels toxic heat and meconium gathered by the fetus. The purpose of this study was to evaluate whether Hataedock alleviates inflammatory skin damage in AD (Atopic Dermatitis)-induced NC/Nga mice through regulating and maintaining the skin barrier and anti-inflammation effects.Methods We established an AD model in three-week-old NC/Nga mice through the repeated application of DNFB (dinitrochlorobenzene) on days 28, 35, and 42 after Hataedock treatment was orally administered. We identified changes in the skin barrier and anti-inflammation effects through the histological and immunohistochemical changes of TNF- α, NF-κB p65, iNOS, COX-2, and apoptotic bodies.Results Skin damage and angiogenesis were mitigated in the HT (Hataedock) group. Damage to the intercellular space of the stratum corneum as well as hyperplasia, edema, the infiltration of lymphocytes, and the increase of capillaries decreased in the HT group. Our results suggest that Hataedock treatment significantly down-regulated levels of TNF- α by 38% (p<0.001) and of NF-κB p65 by 70% (p<0.001). But Hataedock up-regulated apoptosis by 183% in dermatitis-induced skin.Conclusions These results suggest that Hataedock alleviates AD through diminishing the various inflammatory cytokines in skin lesions that are involved in the initial steps of AD development. It might have potential applications for the prevention and treatment of atopic dermatitis.

Effects of Moxi-tar Herbal Acupuncture of LI11 on inflammatory bowel disease induced by TNBS in mice (TNBS로 유도된 대장염에서 곡지의 구진약침 효과)

  • Song, Moon-Young;Park, Sang-Yeon;Kim, Jae-Hyo;Ahn, Sung-Hun;Kim, Kyoung-Sik;Sohn, In-Chul
    • Korean Journal of Acupuncture
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    • v.25 no.3
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    • pp.147-166
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    • 2008
  • Objectives : Crohn's disease (CD) is characterized by a chronic relapsing inflammation of the bowel in which proinflammatory cytokines play an important perpetuating role. Methods : Mice (preventive animal model of gliotoxin) were treated with 5 % 2,4,6-trinitrobenzenesulfonic acid (TNBS) at day 1 and day 7. To investigate preventive effects of acupuncture with Gujin at $LI_{11}$, acupuncture was carried out at day -1, day 1, day 3. And, to investigate therapeutic effects, acupuncture with Gujin was carried out at day 3, day 5, day 7. For the data analysis, we checked weight and width of colon, diarrhea, edema, survival rate, changes of body weight, and myeloperoxygenase (MPO) activity. For analysing protein expression, we carried out immunohistochemical staining and Western blot and we analyzed mRNA expression by RT-PCR. Results : Colon of TNBS treated mice was erosive and shortening compared with the colon of control mice and induced damages of colon epithelial cell layer and induced infiltration of immune cells in all layer of colon. Acupuncture of gujin at $LI_{11}$ in preventive mode suppressed macorscopic damages such as erosive and shortening of colon by TNBS and damages of intestinal epithelial cells and infiltration of immune cells in the colon. The average weight of 5 cm distal colon was increased in TNBS treated mice (758${\mu}g$) compared with in control mice (112${\mu}g$) and width of distal colon was also increased in TNBS treated mice (4.9mm) compared with in control mice (1.3mm). Acupuncture with Gujin at $LI_{11}$ in preventive and therapeutic mode suppressed increase of colon weight and width by TNBS. TNBS induced edema of colon and diarrhea and Acupunctured with Gujin at $LI_{11}$ in preventive and therapeutic mode ameliorated these symptom by TNBS. In preventive and therapeutic mode, the effects of acupuncture with Gujin at $LI_{11}$ were increasing the motility, suppressing body weight decreasing, suppressing MPO activity, reducing expressing of TNF-${\alpha}$, IL-1b, and ICAM-1 in colon compared with that by TNBS Conclusions : This study demonstrates that acupuncture with Gujin at $LI_{11}$ represents a potential therapeutic method of Crohn's disease.

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Morphological Studies on the Localization of Neurons Projecting to the Meridian Points Related to the Facial Nerve Paralysis in the Rat Using the Neural Tracers (신경추적자(神經追跡子)를 이용한 얼굴신경마비(神經痲痺)와 관련(關聯)된 혈(穴)들을 지배(支配)하는 신경세포체(神經細胞體)의 표식부위(標識部位)에 대(對)한 형태학적(形態學的) 연구(硏究))

  • Kim, Jum-Young;Lee, Sang-Ryoung;Lee, Chang-Hyun
    • The Journal of Korean Medicine
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    • v.18 no.1
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    • pp.58-71
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    • 1997
  • In order to the location and local arrangement of nerve cell bodies and nerve fibers projecting to the meridian points related to facial nerve paralysis in the rat using the neural tracers, CTB and WGA-HRP, labeled neurons the were investigated by immunohistochemical and HRP histochemical methods following injection of 2.5% WGA-HRP and 1% CTB into Hyopko$(S_6)$. Chichang$(S_4)$, Sugu$(GV_{26})$, Sajukkong$(TE_{23})$ and Yangbaek$(G_{14})$. Following injection of Hyopko$(S_6)$, Chichang$(S_4)$, labeled motor neurons were founded in facial nucleus, trigeminal motor nucleus, reticular nucleus and hypoglossal nucleus. labeled sensory neurons were founded in trigeminal ganglia and $C_{1-2}$ spinal ganglia. sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in mesencephalic trigeminal tract, sensory root of trigeminal nerve, oral, interpolar and caudal part of trigeminal nucleus, area postrema, nucleus tractus solitarius, lateral reticular nucleus and $C_{1-2}$ spinal ganglia. Following injection of Sugu$(GV_{26})$, labeled motor neurons were founded in facial nucleus. Labeled sensory neurons were founded in trigeminal ganglia and $C_{1-2}$ spinal ganglia. Sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in spinal trigeminal tract, trigeminal motor nucleus, mesencephalic trigeminal tract, oral. interpolar and caudal parts of trigeminal nucleus, area postrema, nucleus tractus solitarius, lateral reticular nucleus, dorsal part of reticular part and $C_{1-2}$ spinal ganglia. Following injection of Sajukkong$(TE_{23})$ and Yangbaek$(G_{14})$, labeled motor neurons were founded in facial nucleus, trigeminal motor nucleus. Labeled sensory neurons were founded in trigeminal ganglia and $C_{1-2}$ spinal ganglia. sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in oral, interpolar and caudal parts of trigeminal nucleus, area postrema, nucleus tractus solitarius, inferior olovary nucleus, medullary reticular field and lamina I-IV of $C_{1-2}$ spinal cord. Location of nerve cell body and nerve fibers projecting to the meridian points related to the facial nerve paralysis in the rats were found in facial nucleus and trigeminal motor nucleus. Sensory neurone were found in trigeminal ganglia and $C_{1-2}$ spinal ganglia. Sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in mesencephalic trigeminal tract, oral, interpolar and caudal parts of trigeminal nucleus, area postrema, nucleus tractus solitarius. lateral reticular nucleus, medullary reticular field.

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THE EXPERIMENTAL STUDY OF THE BONE REGENERATION ON RABBIT MAXILLARY SINUS GRAFTING WITH ${\beta}$-TCP (가토 상악동에 이식된 ${\beta}$-TCP의 골치유에 관한 실험적 연구)

  • Park, Jung-Ha;Hwang, Kyung-Gyun;Park, Chang-Joo;Choi, Yong-Soo;Ma, Pyung-Soo;Paik, Seung-Sam;Shim, Kwang-Sup
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.2
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    • pp.107-116
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    • 2006
  • Purpose:Maxillay sinus grafting is an effective treatment procedure to improve bone height in the posterior maxillar area for implant installation. Beta-tricalciumphosphate(${\beta}$-TCP) was introduced to be grafting substitute material, providing a reasonable bio-degradation time, no need for harvesting procedure. The purpose of this study is to evaluate bone healing & regeneration phase using histomorphometric and immunohistochemical analysis. Material & Methods:Sixteen rabbits were divided into 4 groups. Bi-lateral maxillary sinus membranes were elevated at each rabbits, ${\beta}$-TCP was augmented in left sinus, autogenous bone was augmented in right sinus. The rabbits were sacrificed at 2, 4, 8 and 12 weeks. We investigated the bone regeneration & growth factor expression. Results: 1. The mean new bone volume formation was 28.99${\pm}$6.55%, 49.54${\pm}$5.47%, 69.09${\pm}$8.90% in autogenous grafted area, and 22.86${\pm}$5.56%, 24.00${\pm}$4.09%, 34.11${\pm}$3.37% in ${\beta}$-TCP area at 4, 8, 12 weeks. Therefore, new bone formation in autogenous bone was significantly higher than ${\beta}$-TCP (p<0.05). 2. The BMP 2/4 expression in autogenous bone grafted area was higher at 4, 8 weeks. 3. There was no difference in expression pattern of BMP-7/PDGF/VEGF during grafted bone regeneration. Conclusion:The authors we conclude that the autogenous bone graft was faster than ${\beta}$-TCP in bone regeneration, and the BMP 2/4 were more important in graft bone regeneration.

Immunohistochemical Study on Hair Growth Promoting Effect of Yonnyuniksoogobon-dan (연년익수고본단(延年益壽固本丹)의 육모효능(育毛效能)에 대한 면역조직화학적 연구)

  • Jeong, Chun-Geun;Shim, Eun-Sheb;Lee, Chang-Yeol;Kim, Bum-Hoi;Kim, Seong-Joon;Kang, Hee;Sohn, Nak-Won
    • The Journal of Korean Medicine
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    • v.31 no.2
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    • pp.48-63
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    • 2010
  • Objectives: Yonnyuniksoogobon-dan (Yan Nian Yi Shou Gu Ben Dan 延年益壽固本丹) is composed of 11 herbs (Polygoni Mutiflori Radix, Lycii Radicis Cortex, Polia, Rehmanniae Radix, Rehmanniae Radix Preparat, Asparagi Radix, Liriopis Tuber, Lycii Fructus, Acori Graminei Rhizoma, Angelicae Acutiloba Radix, and Pini Folium) based on Yonryunggobon-dan (Yan Ling Gu Ben Dan 延齡固本丹) and Yonnyuniksoobulrho-dan (Yan Nian Yi Shou Bu Lao Dan 延年益壽不老丹). This study evaluated hair growth promoting effect of Yonnyuniksoogobon-dan on the shaved C57BL/6 mice. Methods: Yonnyuniksoogobon-dan was treated by oral administration (Sample I) and oral administration plus skin application (Sample II) once a day for 12 days. Hair regrowth was photographically and histologically determined during the experimental period. Hair growth cycle related factors (EGF, TGF-${\beta}1$) and vascular factors (VEGF, iNOS) were also determined with immunohistochemistry. Results: 1. On gross observation of hair regrowth, Sample I and Sample II groups demonstrated acceleration of hair regrowth. 2. The hair regrowth index of the Sample I group increased significantly from 7 days (P<0.05) to 12 days (P<0.01) after the shave while that of the Sample II group significantly increased at 12 days (P<0.05). 3. On histological observation, both Sample I and Sample II groups demonstrated histological improvement and increases of number and diameter of the hair follicles. 4. EGF expressions on the root sheath of hair follicles were up-regulated in both Sample I and Sample II groups. 5. TGF-${\beta}1$ expressions on the root sheath of hair follicles were not regulated in Sample I or Sample II groups. 6. VEGF expressions in the surrounding tissues of hair follicles were up-regulated in both Sample I and Sample II groups. 7. iNOS expressions in the surrounding tissues of hair follicles were down-regulated in both Sample I and Sample II groups. Conclusions: These results suggest that Yonnyuniksoogobon-dan has hair growth-promoting activity and these effects relate to up-regulations of EGF and VEFG expressions and down-regulations of TGF-${\beta}1$ and iNOS expressions on hair roots.

Immunohistochemical Study on the Hair Growth Promoting Effect of Yonnyuniksoogobon-dan

  • Jeong, Chun-Geun;Park, Min-Hee;Seong, Ju-Won;Lee, Hyun-Sam;Park, Seong-Kyu;Kim, Sun-Yeou;Kim, Yoon-Bum;Jung, Hyuk-Sang;Sohn, Nak-Won;Sohn, Young-Joo
    • The Journal of Korean Medicine
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    • v.29 no.5
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    • pp.77-89
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    • 2008
  • Objectives : This study evaluates the hair growth promoting effect of Yonnyuniksoogobon-dan on shaved C57BL6 mice. Methods : Yonnyuniksoogobon-dan was administered orally (Group I) and both orally and by skin application (Group II) once a day for 12 days. The experimental groups were compared to Control, which was orally administered physiological saline solution. Hair regrowth was photographically and histologically determined during the experimental period. The levels of hair growth cycle related factors (EGF, TGF-${\beta}$1) and vascular factors (VEGF, iNOS) were also determined by immunohistochemistry. On gross observation of hair growth, both Group I and Group II shaved C57BL6 mice showed accelerated hair regrowth. Results : The hair regrowth index of Group I increased significantly from 7 days (P<0.05) to 12 days (P<0.01) after shaving and that of Group II was significantly higher at 12 days (P<0.05). On histological observation, both Group I and Group II demonstrated histological improvement and increases in the number and diameter of the hair follicles. EGF expression on the root sheath of hair follicles was up-regulated in both Group I and II. TGF-${\beta}$1 expression on the root sheath of hair follicles was unchanged in both Group I and II. VEGF expression in the tissues surrounding hair follicles was up-regulated in both groups. iNOS expression in the tissues surrounding hair follicles was down-regulated in both groups. Conclusions : These results suggest that Yonnyuniksoogobon-dan promotes hair growth and this effect is related to up-regulation of EGF and VEFG expression and down-regulation of TGF-${\beta}$1 and iNOS expression on hair roots.

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Effect of Kami-chungsimyeunjatang on atopic dermatitis-like skin lesions induced in NC/Nga mice by mite antigen stimulation (가미청심연자탕(加味淸心蓮子場)이 NC/Nga mice의 아토피양(樣) 피부염에 미치는 영향)

  • Han, Jae-Kyung;Kim, Yun-Hee;Yoon, Ji-Yeon
    • The Journal of Pediatrics of Korean Medicine
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    • v.21 no.1
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    • pp.87-116
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    • 2007
  • Objectives : The purpose of this study is to examine of the effect of Kami-chungsimyeunjatang(KCSYJT) medicine on the atopy eruption control. Methods : The expression of IgE, IL-4, IL-6, $TNF-{\alpha}$, IgG2b, IgM, IgG2a and IgG1 level in serum, and $IFN-{\gamma}$ production by KCSYJT were analyzed. CD3e+/CD69+, CD4+/CD25+, B220+/IgE+ and B220+/CD23+ positive cells by flow cytometry in splenocytes were assayed and the revelation of CD3e+/CD69+, CD4+/CD8+ and CD4+/CD25+ marker in PBMC, spleen and DLN were observed. The outturn of IL-4, eotaxin 2, CCR3, TARC mRNA in splenocytes werw observed. We also analyzed NC/Nga mice's ear, DLN and neck-back skin after biopy and dye by H&E, and toluidine staining (mast cells marker) method, measured about epidermis and dermis part in comparison with control group. Results : NC/Nga mice suffered from dermatitis very similar to human AD with IgE hyperproduction. Specially, result that measure IgE content in serum on 8 weeks, 12 weeks, 16 weeks decreased remarkably than control group. After experiment end, result that observe revelation CD3e+/CD69+, CD4+/CD8+ and CD4+/CD25+ marker in PBMC, spleen and DLN establishment observed recover as normal with political background. And decreased than result control group which measure IL-4, IL-6, $TNF-{\alpha}$, IgG2b, IgM, IgG2a, IgG1 level in serum, and $IFN-{\gamma}$ production secreted in Th1 cell displayed increase by KCSYJT medicines. Ear thickness decrease than control group in result that observe effect that get in ear of a NC/Nga mouse. Course inflammation immunocyte etc.. permeated of result that effect that KCSYJT medicines get to NC/Nga mouse's skin establishment analyzes ear, DLN and neck-back skin after biopy, and dye by H&E, and toluidine staining (mast cells marker) method decreased about epidermis. and inflammation of dermis part remarkably than control group. Immunohistochemical examination of the skin lesion showed decrease by KCSYJT medicines on numbers of mast cells (CCR3) and CD4+ T cells containing IL-4 necessary for IgE. Conclusions : Th1 cell and Th2 cell was observed to be shift by secretion amount of IL-4 and $IFN-{\gamma}$ by KCSYJT medicines. Therefore, the KCSYJT medicine turned out to be useful in allergy autoimmune disease.

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Functional Connections of the Vestibulo-spino-adrenal Axis in the Control of Blood Pressure Via the Vestibulosympathetic Reflex in Conscious Rats

  • Lu, Huan-Jun;Li, Mei-Han;Li, Mei-Zhi;Park, Sang Eon;Kim, Min Sun;Jin, Yuan-Zhe;Park, Byung Rim
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.5
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    • pp.427-434
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    • 2015
  • Significant evidence supports the role of the vestibular system in the regulation of blood pressure during postural movements. In the present study, the role of the vestibulo-spino-adrenal (VSA) axis in the modulation of blood pressure via the vestibulosympathetic reflex was clarified by immunohistochemical and enzyme immunoassay methods in conscious rats with sinoaortic denervation. Expression of c-Fos protein in the intermediolateral cell column of the middle thoracic spinal regions and blood epinephrine levels were investigated, following microinjection of glutamate receptor agonists or antagonists into the medial vestibular nucleus (MVN) and/or sodium nitroprusside (SNP)-induced hypotension. Both microinjection of glutamate receptor agonists (NMDA and AMPA) into the MVN or rostral ventrolateral medullary nucleus (RVLM) and SNP-induced hypotension led to increased number of c-Fos positive neurons in the intermediolateral cell column of the middle thoracic spinal regions and increased blood epinephrine levels. Pretreatment with microinjection of glutamate receptor antagonists (MK-801 and CNQX) into the MVN or RVLM prevented the increased number of c-Fos positive neurons resulting from SNP-induced hypotension, and reversed the increased blood epinephrine levels. These results indicate that the VSA axis may be a key component of the pathway used by the vestibulosympathetic reflex to maintain blood pressure during postural movements.

Beneficial Effect of the Combination of Oral Administration and Herbal -Acupuncture Stimulation with Several Herb-combind Prescription on Streptozotocin-Induced Diabetic Rats (한약복합처방(韓藥複合處方) 약침(藥鍼) 및 경구투여(經口投與)가 Streptozotoin에 의한 흰쥐의 당뇨병(糖尿病)과 항산화능(抗酸化能)에 미치는 영향)

  • Park, Sa-hyun;Cho, Su-in;Chae, Woo-seok;Cho, Myung-rae
    • Journal of Acupuncture Research
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    • v.22 no.1
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    • pp.1-11
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    • 2005
  • Objective : The present study was carried out to investigate the preventive effect of Several Herb-combind Prescription(SHP) on Streptozotocin (STZ) -induced Diabetes mellitus. Methods : SHP was given to rats with the combination of oral administration and herbal-acupuncture stimulation. The experimental animals were divided into 3 groups : normal group of rats, control group of STZ-induced diabetic rats, sample group with SHP treatment. In vitro test of SHP showed ${\alpha}$-glucosidase inhibition, DPPH radical scavenging activity and inhibition of lipid peroxidation. Experimental diabetes was induced by the injection of STZ(60mg/kg) to the rat via the peritoneum. The effect of SHP on STZ-induced diabetes was observed by measuring the seum level of insulin, glucose, triglyceride, total cholesterol and lipid peroxides. Hepatic activities of catalase and reduced glutathione were examined and insulin granule was observed by immunohistochemical examination. Result : STZ caused hyperglycemia and hypoinsulinemia by a selectively destroying pancreatic ${\beta}$-cell. SHP treatment protected them from the hyperglycemia and hypoinsulinemia. STZ induced increase of serum triglyceride lowered by SHP treatment. And by SHP treatment, pancrease showed a big area with positive immuno-reactivity for presence of insulin with many insulin granules distributed in the ${\beta}$-cells in the islets of Langerhans. Contusions : The SHP treatment showed protective effect on diabetic rat model, and action mechanism of the effect was thought to be concerned with anti-oxidative stress.

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