• Title/Summary/Keyword: $4{\beta}$

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감마선 조사에 의한 뇌조직의 Jun 및 p53유전자 발현 (Expression of Jun and p53 Genes from the Brain of Rats Irradiated with $^{60}Co{\gamma}$-ray)

  • 김용석;우종규;이용성;고재경;전하정;이명자
    • Radiation Oncology Journal
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    • 제14권4호
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    • pp.265-279
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    • 1996
  • 방사선은 DNA손상을 초래하고 세포 성장에 관련된 유전자의 발현 조절 및 apoptosis등을 유발한다고 알려져 있으며 본 연구는 신경계에 있어 방사선 조사 후 종양 발생율과 시간 경과의 관계 및 조사 양과 암 발생의 관계를 구명하기 위해 코발트 60의 전신조사에 따른 흰쥐 뇌 조직의 생체 반응을 연구하고자 하였다. 이를 위하여 상기조직의 jun 및 p53 유전자의 발현도를 1 Gy로 부터 100 Gy 범위의 감마선 용량별 및 1시간에서 6시간까지의 조사 훈 경과 시간 별로 Northern 분석하였다. Jun유전자 발현도는 ley이하에서 1시간 이내에 한계수준에 도달하였으며 30 Gy의 조사 1시간 째에 최대였다. 또한 조사 1시간 이후 1 Gy로부터 10 Gy 범위에서는 조사 5시간 및 6시간까지 점진적으로 증가되었으나 20 Gy로부터 100 Gy 범위에서는 조사 2시간까지 증가 후 감소되는 양상을 나타냈다. p53유전자의 발현도는 1 Gy이하에서 1시간 이내에 한계 수준에 근접했고 1 Gy의 조사 후 6시간 째에 최대였다. 1 Gy로부터 40 Gy까지의 범위에서는 조사 5시간 및 6시간까지 점진적으로 증가되는 반면 50 Gy에서 100 Gy범위에서는 조사 2시간 째까지 증가 후 감소되는 양상을 보였다. 따라서 감마선 조사양이 높을수록 jun 및 p53유전자는 신속하게 최대로 발현되었고 감마선 조사양이 낮을수록 서서히 증가되었다 그러나 jun유전자와 p53유전자의 감마선 조사에 따른 발현 양상에는 상호간의 연관성을 찾을 수 없었다.

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노루궁뎅이균사발효 율무 열수추출물의 유용성분 및 생리활성 (Biological activity in hot water extract from fermented Coix lacryma-jobi L. var. mayuen Stapf. by Hericium erinaceum (Bull. : Fr.))

  • 윤경원;김경제;진성우;고영우;임승빈;하늘이;정희경;정상욱;서경순
    • 한국버섯학회지
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    • 제18권1호
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    • pp.20-28
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    • 2020
  • 노루궁뎅이균사로 발효한 율무 열수추출물의 항산화, NO 생성저해, tyrosinase 합성 억제, melanin 생성 저해 및 유용성분 분석을 수행하였다. 노루궁뎅이균사발효율무열수추출물의 ergosterol 함량은 740.2 mg%으로 나타났으며, 원료로 사용한 율무 열수추출물에서는 검출되지 않았다. 노루궁뎅이균사발효율무 열수추출물의 β-glucan 함량은 245.3 ± 5.1 mg%으로 나타났다. 노루궁뎅이균사발효율무 열수추출물은 100%, 104.1%, 107.2%, 105.3%, 102.1%, 101.3%, 100.4%의 세포생존율이 측정되어, 율무열수추출물보다 노루궁뎅이균사발효율무 열수추출물의 세포생존율이 더 높게 나타났다. B16F10 cell 500 ug/mL 농도로 처리했을 때, 노루궁뎅이균사발효율무 열수추출물은 8.9 μM의 NO 생성율을 보여, 율무 추출물(10.6 uM)의 NO생성율 보다 낮은 NO 생성율을 나타내었다. 10, 30, 50, 100, 200, 300 및 500 mg/mL 모든 농도에서 율무 열수추출물보다 노루궁뎅이균사발효율무 열수추출물이 유의적으로 tyrosinase 및 melanin 생성을 억제함을 확인 할 수 있었다. 노루궁뎅이균사발효에 따른 DPPH radical 및 ABTS radical 소거활성 측정결과 노루궁뎅이 균사발효 율무열수추출물이 율무 열수추출물보다 항산화 효과가 높음을 확인하였다. 따라서 추가적인 연구를 통해 노루궁뎅이균사발효 율무열수추출물은 기능성식품 원료와 식품 첨가물, 화장품 산업에 이용될 수 있을 것으로 생각된다.

미세조작조건이 소 핵이식배의 발달에 미치는 영향 (Effects of Manipulation Conditions on Development of Nuclear Transplant Bovine Embryos Derived from In Vitro Matured Oocytes)

  • 최상용;노규진;공일근;송상현;조성근;박준규;이효종;박충생
    • 한국가축번식학회지
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    • 제21권3호
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    • pp.293-302
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    • 1997
  • Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

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아말감의 구강내 부식 및 인공 부식에 관한 연구 (A STUDY ON IN VIVO AND IN VITRO AMALGAM CORROSION)

  • 임병목;권혁춘;엄정문
    • Restorative Dentistry and Endodontics
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    • 제22권1호
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    • pp.1-33
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    • 1997
  • The objective of this study was to analyze the in vitro and in vivo corrosion products of low and high copper amalgams. The four different types of amalgam alloy used in this study were Fine cut, Caulk spherical, Dispersalloy, and Tytin. After each amalgam alloy and Hg were triturated according to the directions of the manufacturer by means of the mechanical amalgamator(Amalgam mixer. Shinhung Co. Korea), the triturated mass was inserted into a cylindrical metal mold which was 12mm in diameter and 10mm in height. The mass was condensed by 150Kg/cm compressive force. The specimen was removed from the mold and aged at room temperature for about seven days. The standard surface preparation was routinely carried out by emery paper polishing under running water. In vitro amalgam specimens were potentiostatically polarized ten times in a normal saline solution at $37^{\circ}C$(potentiostat : HA-301. Hukuto Denko Corp. Japan). Each specimen was subjected to anodic polarization scan within the potential range -1700mV to+400mV(SCE). After corrosion tests, anodic polarization curves and corrosion potentials were obtained. The amount of component elements dissolved from amalgams into solution was measured three times by ICP AES(Inductive Coupled Plasma Atomic Emission Spectrometry: Plasma 40. Perkim Elmer Co. U.S.A.). The four different types of amalgam were filled in occlusal and buccal class I cavities of four human 3rd molars. After about five years the restorations were carefully removed after tooth extraction to preserve the structural details including the deteriorated margins. The occlusal surface, amalgam-tooth interface and the fractured surface of in vivo amalgam corrosion products were analyzed. In vivo and in vitro amalgam specimens were examined and analyzed metallographically by SEM(Scanning Electron Microscope: JSM 840. Jeol Co. Japan) and EDAX(Energy Dispersive Micro X-ray Analyser: JSM 840. Jeol Co. Japan). 1. The following results are obtained from in vitro corrosion tests. 1) Corrosion potentials of all amalgams became more noble after ten times passing through the in vitro corrosion test compared to first time. 2) After times through the test, released Cu concentration in saline solution was almost equal but highest in Fine cut. Ag and Hg ion concentration was highest in Caulk spherical and Sn was highest in Dispersalloy. 3) Analyses of surface corrosion products in vitro reveal the following results. a)The corroded surface of Caulk spherical has Na-Sn-Cl containing clusters of $5{\mu}m$ needle-like crystals and oval shapes of Sn-Cl phase, polyhedral Sn oxide phase. b)In Fine cut, there appeared to be a large Sn containing phase, surrounded by many Cu-Sn phases of $1{\mu}m$ granular shapes. c)Dispersalloy was covered by a thick reticular layer which contained Zn-Cl phase. d)In Tytin, a very thin, corroded layer had formed with irregularly growing Sn-Cl phases that looked like a stack of plates. 2. The following results are obtained by an analysis of in vivo amalgam corrosion products. 1) Occlusal surfaces of all amalgams were covered by thick amorphous layers containing Ca-P elements which were abraded by occlusal force. 2) In tooth-amalgam interface, Ca-P containing products were examined in all amalgams but were most clearly seen in low copper amalgams. 3) Sn oxide appeared as a polyhedral shape in internal space in Caulk spherical and Fine cut. 4) Apical pyramidal shaped Sn oxide and curved plate-like Sn-Cl phases resulted in Dispersalloy. 5) In Tytin, Sn oxide and Sn hydroxide were not seen but polyhedral Ag-Hg phase crystal appeared in internal space which assumed a ${\beta}_l$ phase.

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간기능강화제의 수침이 간손상 유발견의 회복에 미치는 영향 (The Effect of Aquapuncture with Hepatonics on the Recovery in Artificially Induced Hepatic Damaged Dogs)

  • 유명조;조성환;윤원기;김덕환;유기덕
    • 한국임상수의학회지
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    • 제14권2호
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    • pp.308-318
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    • 1997
  • The present experiment was performed in order to know the treatment effect of aquapuncture with hepatonics on recovery in artificially induced hepatic damaged dogs by carbon tetrachloride. The animals were divided into a control and two experimental groups (aquapuncture with taurine into Gan-su acupoint: Aa-I group and aquapuncture with taurine into blank acupoint in the thigh: Aa-II group). The changes of serum enzyme activities (ALT, AST and ALP), serum total protein contents, protein fractions and pathohistological findings of the liver were examined after application of aquapunctiuf treauent The results obtained through this experiment were summarized as follows : The serum ALT activities tendeded to decrease in experimental group compared with those of control group. Significances were detected at 5th (p<0.05) and 7th (p<0.05) day in Aa-I group and 7th day (p<0.05) in Aa-II group, respectively. Low value was fecund in Aa-I group compared with Aa-II groups but significance was not observed between two experimental groups. The serum AST activities in experimental group showed decreasing tendency compared with those of control group. Significances were observed at 2nd (p<0.05) and 5th (p<0.05) day in Aa-I group and 2nd (p<0.05) day in Aa-II groups respectively. Aa-I group showed lower values than those of Aa-II groups however, no significance was detected between experimental groups. The serum ALP activities of experimental group showed a slight decrease compared with those of control group, however, significance was not detected among all groups. The serum total protein content in experimental group showed tendency of increase compared with control group. Significance was found at 2nd day (p<0.05) in Aa-I group, but there was no significance in Aa-II group. Further significant increase of total protein content was seen at 1st day (p<0.05) in Aa-I group compared with Aa-II group. The change of serum albumin content in experimental group showed tendency of increase compared with control. Significant increases were detected at 1st (p<0.01) and 2nd (p<0.01) day in Aa-I group, respectively. Aa-II group showed increase compared with control groups but significance was not observed. Further significant increase was at 1st day (p <0.05) in Aa-I group compared with Aa-II group. The change of ${\beta} $-globulin in Aa-I group was slightly decreased compared with control group. Aa-II group was similar to controls but significance was not observed among all groups. The change of P-globulin content in Aa-I group showed tendency of increase compared with control and Aa-II group showed the tendency of decrease compared with control. The change of ${\gamma}$-globulin content experimental group showed tendency of increase compared with control, however, significance was not detected among groups. The change of A/G ratio in Aa-I group showed tendency of increase compared with control group and Aa-II group was similar to controls but no significance was found among groups. As for pathohistological observations the grade of hepatocellular vacuolized degeneration and necrosis in Aa-I group was milder than those of control and Aa-II groups and the change of Aa-II group was similar to that of control. Considering above finding collectively, it was thought that aquapuncture of Gan-su acupoint with hepatonics was more effective than aquapuncture of blank acupoint for the recovery of hepatic damage.

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랫트에서 Chitosan-Trimer가 복강유착에 미치는 영향 (Effect of Chitosan-Trimer on the Prevention of Postoperative Intraperitoneal Adhesion Formation in Rats)

  • Kwon, Eun-ju;Jang, Kwang-ho;Jang, In-ho
    • 한국임상수의학회지
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    • 제18권3호
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    • pp.257-264
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    • 2001
  • 복강 내 유착은 단층편평상피로 구성된 복막의 손상으로 장막표면의 염증반응을 일으켜 창상조직의 혈관투과성이 증가하여 많은 장액성 혈액삼출물이 생산되고, 이 혈액삼출물내의 fibrin이 제거되지 않으면 초기섬유소성 유착이 발생한다. 따라서 유착방지는 섬유소성 부착물의 정상적인 용해를 방해하는 인자들과 관계가 있다. Chitosan은 poly-$\beta$(1$\longrightarrow$4)-D-glucosamine으로 chitin을 탈아세틸화시킨 것으로 복강유착방지에 효과가 있다고 알려진 Hyaluronic acid(HA)와 구조상 유사성을 가지고 있다. 본 연구는 쥐에서 회장에 유착을 유도한 후 PBS (control group), 1% Chitosan Trimer (CT, 1% CT group), 3% CT (3% group), chitin (chitin group)을 복강내 주입하여서 10일 뒤에 유착방지효과, 유착발생정도, 조직검사, 혈액상의 변화를 관찰하였다. 총백혈구수, 총적혈구수, PCV, PLT, Total protein은 전군에서 유의적인 변화가 나타나지 않았다. 조직검사상에서 유의적인 차이는 나타나지 않았으나, 3% CT군은 다른 군에 비해 Fibrosis와 염증반응정도에 대한 점수가 낮았다. 혈장섬유소원은 전군에서 수술 후 증가하였으나 3% CT군은 대조군에 비해 증가율이 낮아서 유의적인 차이를 보였다 (p<0.05). 유착장소는 전군에서 장막-장막 (60%), 장막-장간막 (13.3%), 장막-고환쪽 결합조직 (10%), 대망-간 (10%), 장막-대망 (3.3%), 장막-맹장3 (3.3%)순으로 발생하였다. 유착발생빈도는 3% CT군이 62.2% 로 대조군 97.7%, 1% CT군 81.8%, chitin군 93.3%에 비해 유의적으로 낮았다 (p<0.05). 유착 형성은 대조군, 1% CT, 3% CT 및 chitin 투여군에서 각각 2.07$\pm$0.81, 1.03$\pm$0.63, 0.64$\pm$0.53 및 1.67$\pm$0.71로 3% CT군은 대조군에 비해 유의적인 감소를 보였다 (p<0.001).

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강황과 비트를 첨가한 젤리의 품질특성 (Quality Characteristics of Jelly Containing Added Turmeric (Curcuma longa L.) and Beet (Beta vulgaris L.))

  • 조영;최미용
    • 한국식품조리과학회지
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    • 제26권4호
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    • pp.481-489
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    • 2010
  • 본 연구에서는 강황과 비트의 기능성을 살린 젤리를 실용화시키고 효과적인 배합비를 알아보기 위해 강황과 비트 분말의 첨가 비율을 달리해서 젤리를 제조한 후 pH, 산도, 당도, 색도, 투명도, 탁도, 물성, 관능검사를 실시하여 젤리의 품질특성을 알아보았다. 강황 젤리는 강황 분말 첨가량이 증가할수록 pH와 당도가 유의적으로 높아지고(p<0.001), 산도는 낮아졌다(p< 0.001). 비트 젤리는 비트 분말 첨가량이 증가할수록 pH가 유의적으로 낮아졌고(p<0.001), 산도와 당도는 유의적으로 증가하였다(p<0.001). 색도는 강황 젤리의 경우 강황 분말 첨가량이 증가할수록 명도(L)값은 유의적으로 감소하였고, 황색도(b)값과 적색도(a)값은 유의적으로 증가하는 경향을 보였다(p<0.001). 그리고 비트 젤리는 비트 분말의 첨가량이 증가할수록 명도(L)값과 황색도(b)값은 감소했고, 적색도(a)값은 유의적으로 증가하는 경향을 보였다(p<0.001). 투명도는 강황 젤리와 비트 젤리 모두 분말 첨가량이 증가할수록 유의적으로 감소하는 경향을 보였으나, 탁도는 유의적으로 증가하였다(p<0.001). 물성은 강황 젤리의 경우 강황 분말 첨가량이 증가함에 따라 견고성과 탄력성은 낮아졌고, 부착성은 증가하였다. 응집성은 아주 서서히 감소하였으며 유의적 차이는 없었다. 검성도 강황 분말 첨가량이 증가함에 따라 낮아졌으며, 씹힘성은 대조군보다 현저하게 낮아졌다. 또한 비트 젤리는 비트 분말 첨가량이 증가할수록 견고도는 증가하였으나 유의적 차이는 없었다. 부착성은 비트 분말 첨가량이 증가할수록 유의적으로 증가하는 경향을 보였고(p<0.001), 탄력성도 유의적으로 낮아졌으며(p<0.05), 응집성은 낮아졌으나 유의적 차이는 없었다. 검성은 비트 분말이 첨가되면서 낮아졌다. 씹힘성은 비트 분말의 첨가량 증가에 따라 유의적으로 낮아졌다(p<0.001). 관능검사 결과는 강황 분말 첨가 젤리의 경우 색상은 T1(0.5%)가 가장 높은 값을 보였고, 외관과 단맛도 T1(0.5%)가 가장 높았다(p<0.001). 씹힘성, 탄력성, 견고성, 투명도, 전반적인 수용도는 대조군이 모두 높았으나 그 다음으로는 T1(0.5%)가 높았다. 또한 비트 분말 첨가 젤리의 경우도 색상, 외관, 씹힘성, 탄력성은 B2(1%)가 가장 높은 값을 보였고, 단맛은 대조군이 조금 높았으나, 약간의 차이가 있었다. 견고성, 투명도는 대조군이 모두 높았으나 그 다음으로 B1(0.5%)가 높은 값을 나타냈다. 전반적인 수용도는 B2(1%)가 가장 높았다. 따라서 강황과 비트의 기능성을 살린 젤리를 실용화시키는 효과적인 배합비는 강황 분말의 경우 0.5%, 비트 분말은 1% 첨가가 적절한 것으로 나타났다. 그러므로 강황과 비트는 천연 색소와 기능성을 포함한 식품 소재로서 젤리에 강황과 비트 분말을 첨가 하는 것은 강황과 비트를 활용할 가치와 효과가 있다고 사료된다.

인회석 박막 피복 도관과 Brain-derived neurotrophic factor(BDNF) 유전자 이입 슈반세포를 이용한 백서 좌골신경 재생에 관한 연구 (SCIATIC NERVE REGENERATION USING CALCIUM PHOSPHATE COATED CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE-TRANSFECTED SCHWANN CELL IN RAT)

  • 최원재;안강민;황순정;정필훈;김명진;김남열;유상배;장정원;김현만;김중수;김윤희;김성민;이종호
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제31권3호
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    • pp.199-218
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    • 2005
  • Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

Anti-diabetic effect and mechanism of Korean red ginseng extract in C57BL/KsJ db/db mice

  • ;;정성현
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 2007년도 추계 학술대회
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    • pp.57-58
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    • 2007
  • Purpose: Ginseng is a well-known medical plant used in traditional Oriental medicine. Korean red ginseng (KRG) has been known to have potent biological activities such as radical scavenging, vasodilating, anti-tumor and anti-diabetic activities. However, the mechanism of the beneficial effects of KRG on diabetes is yet to be elucidated. The present study was designed to investigate the anti-diabetic effect and mechanism of KRG extract in C57BL/KsJ db/db mice. Methods: The db/db mice were randomly divided into six groups: diabetic control group (DC), red ginseng extract low dose group (RGL, 100 mg/kg), red ginseng extract high dose group (RGH, 200 mg/kg), metformin group (MET, 300 mg/kg), glipizide group (GPZ, 15 mg/kg) and pioglitazone group (PIO, 30 mg/kg), and treated with drugs once per day for 10 weeks. During the experiment, body weight and blood glucose levels were measured once every week. At the end of treatment, we measured Hemoglobin A1c (HbA1c), blood glucose, insulin, triglyceride (TG), adiponectin, leptin, non-esterified fatty acid (NEFA). Morphological analyses of liver, pancreas and white adipose tissue were done by histological observation through hematoxylin-eosin staining. Pancreatic islet insulin and glucagon levels were detected by double-immunofluorescence staining. To elucidate an action of mechanism of KRG, DNA microarray analyses were performed, and western blot and RT-PCR were conducted for validation. Results: Compared to the DC group mice, body weight gain of PIO treated group mice showed 15.2% increase, but the other group mice did not showed significant differences. Compared to the DC group, fasting blood glucose levels were decreased by 19.8% in RGL, 18.3% in RGH, 67.7% in MET, 52.3% in GPZ, 56.9% in PIO-treated group. With decreased plasma glucose levels, the insulin resistance index of the RGL-treated group was reduced by 27.7% compared to the DC group. Insulin resistance values for positive drugs were all markedly decreased by 80.8%, 41.1% and 68.9%, compared to that of DC group. HbA1c levels in RGL, RGH, MET, GPZ and PIO-treated groups were also decreased by 11.0%, 6.4%, 18.9%, 16.1% and 27.9% compared to that of DC group, and these figure revealed a similar trend shown in plasma glucose levels. Plasma TG and NEFA levels were decreased by 18.8% and 16.8%, respectively, and plasma adiponectin and leptin levels were increased by 20.6% and 12.1%, respectively, in the RGL-treated group compared to those in DC group. Histological analysis of the liver of mice treated with KRG revealed a significantly decreased number of lipid droplets compared to the DC group. The control mice exhibited definitive loss and degeneration of islet, whereas mice treated with KRG preserved islet architecture. Compared to the DC group mice, KRG resulted in significant reduction of adipocytes. From the pancreatic islet double-immunofluorescence staining, we observed KRG has increased insulin production, but decreased glucagon production. KRG treatment resulted in stimulation of AMP-activated protein kinase (AMPK) phosphorylation in the db/db mice liver. To elucidate mechanism of action of KRG extract, microarray analysis was conducted in the liver tissue of mice treated with KRG extract, and results suggest that red ginseng affects on hepatic expression of genes responsible for glycolysis, gluconeogenesis and fatty acid oxidation. In summary, multiple administration of KRG showed the hypoglycemic activity and improved glucose tolerance. In addition, KRG increased glucose utilization and improved insulin sensitivity through inhibition of lipogenesis and activation of fatty acid $\beta$-oxidation in the liver tissue. In view of our present data, we may suggest that KRG could provide a solid basis for the development of new anti-diabetic drug.

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생약복합물에 의한 지방세포형성 조절자의 유전자 발현 연구 (A Study on the Gene Expression of Adipogenic Regulators by an Herbal Composition)

  • 이해용;강련화;배성민;채수안;이정주;오동진;박석원;조수현;심예지;윤유식
    • 생명과학회지
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    • 제20권5호
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    • pp.729-735
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    • 2010
  • 본 연구의 목적은 생약복합제제인 SH21B의 adipogenesis 억제 효능에 대한 상세한 분자적 메커니즘을 3T3-L1 지방세포를 이용하여 밝히는 데 있다. 실험에 사용된 SH21B는 7가지 생약성 천연물질인, 황금, 행인, 마황, 석창포, 포황, 원지 및 하엽으로 이루어졌다. 최근, 본 연구진에 의해 3T3-L1을 이용한 in vitro 연구와 마우스를 이용한 in vivo 연구에서 SH21B의 adipogenesis 억제효능이 밝혀진 바 있다. 본 연구에서는 3T3-L1 지방세포가 분화될 때 작용하는 다양한 지방세포형성 조절자들의 유전자 발현이 SH21B에 의해 어떻게 변하는지 살펴보고자 하였다. 실시간중합효소반응(real time PCR) 기술을 이용하여 SH21B를 처리한 지방세포와 그렇지 않은 지방세포를 비교한 결과, 최종마커인 ADIPOQ와 SLC2A4의 유전자 발현이 SH21B에 의해 급격하게 감소함을 알 수 있었다. 최종마커의 발현을 유도하는 핵심전사인자인 $PPAR{\gamma}$와 C/$EBP{\alpha}$의 유전자 발현 역시 SH21B의 처리 시 유의하게 억제되었다. 좀 더 상세한 분자적 메커니즘을 규명하기 위해, 핵심전사인자의 상위에 위치한 다양한 조절자들의 유전자 발현을 분석하였다. 그 결과, 여러 지방세포형성 유도조절자 중, Krox20과 KLF15의 유전자 발현이 SH21B 처리에 의해 유의하게 감소된 반면, C/$EBP{\beta}$와 KLF5의 유전자 발현은 SH21B 처리에 영향을 받지 않았다. 그리고 지방세포형성 억제조절자인 KLF2와 CHOP의 유전자 발현은 SH21B 처리에 의해 유의하게 증가되었다. 이러한 결과들은 SH21B의 지방세포형성 억제효능이 지방세포의 분화에 작용하는 다양한 상위조절자 중 지방세포형성 유도조절자인 Krox20과 KLF15 그리고 지방세포형성 억제조절자인 KLF2와 CHOP 등의 유전자 발현이 변화되면서 일어나는 복합적인 반응의 결과임을 제시한다.