모델동물의 병태.생리 유발 유전자 기능 연구 : 당뇨모델동물의 췌장 β-세포를 이용한 비만발생에 관한 연구 Effect of development of diabetes by fatty acid in animal model

황대연,김용규,강태석,황진희,임채형,강현구,이주은,허영범,장인석,조정식,채갑용 식품의약품안전청 2001 식품의약품안전청 연보 Vol.5 No.-

체장 β세포에 지방이 축적되는 lopotoricity 현상은 인슬린 비의존성 당뇨 (Non InsuBin Dependent Diabetes Mellitus :NIDDM)의 발생과 상당히 밀접한 관계가 있으며, 췌장 β세포의 lipoapoptosis는 세포막형성에 관여하는 sphingomyelin의 전군체안 ceramide의 과생산에 의해 유도된다는 보고가 있다. 그러나, 지방산 종류에 따른 췌장 β세포에서의 세포사멸 (apoptosis)유발 기작에 대해서는 야직까지 보고된 바가 없파. 또한, 지방세포에서 생성되며, 지질대사, 음식섭취 및 체중조절에 관여하는 렙틴 (leptin)과 렙틴 수용체와의 관계 역시 체장 세포에서 명착하게 밝혀져 있지 않다. 따라서 본 연구에서는 6주령 53와 Zucker rat의 췌장으로부터 췌도를 분리하는 기술을 확립하고, 배양된 췌장 췌도 세포와 MS-1세포주에 불포차즌방산 (n-3 linolenic acid; n-6 :linoleic acid), 포화지방산 (stearic acid), conjugated linoleic acid (CLA)를 포함하는 4 가지의 지방산을 처리한 후 이들이 췌장 』세포의 사멸에 미치는 영향을 다양한 apoptosis 관련 유전자의 발현을 분석함으로써 확인하고자 하였다. 그 결과 SD rat으로부터 마리 당 평균 약 500개의 췌장 췌도를 분리하는데 성공하였으며, 4 종류의 지방산 중 포화지방산인 stearic acid를 처리하여 ONA fragmentation 증가를 관찰하였다. 또한 RT-PCR 방법을 이용하여 분리된 췌도 세포에사의 Bcl-2, Bax, iNOS가 지방산의 종류에 따라 서료 다른 발현양상을 나타낸다는 」라실을 관귿하였다. 이러한 떤구 결과는 지방산의 종류에 따라 췌장 췌도 세포에서의 세포사멸 유발 효과를 겋증하였으며, 이러한 연구결과는 복잡한 당뇨발병 원인 규명에 중요한 정보를 제공할 것으로 사료된다. Fat contents within the pancreatic islets are closely related to lipotoxicity which induces apoptosis, resulting in non insulin dependent mellituse(NIDDM). High level of fat may contribute to both insulin resistance and obesity in rodent and humans but the evidence is limited. To determine whether dietary fatty acids affect pancreatic cell function, the INS-1 cells and the pancreatic islets isolated from SD and Zucker obese(fa/fa) rats(6 wk, prediabetic stage) were cultured with stearic acid and CLA. As a result, DNA fragmentation laddering was substantially decreased in the INS-1 cells and the isolated pancreatic islets cultured with 2mM CLA compared with those with stearic aced. To investigate the mechanism by which CLA alleviates cell apoptosis as indicated by DNA fragmentation assay, we examined mRNA expressions of apoptosis-related proteins including Bax and Bcl-2 associated with cell death agonist and antagonist, respectively in the both INS-1 cells and islets cultured with 2 mM fatty acids. Bax mRNA expression was not markedly altered by either stearic acid or CLA, whereas Bcl-2 mRNA expression was noticeably enhanced by CLA compared with stearic acid. However, there were no changes in cell apoptosis and cell apoptotic-regulating gene products in the both INS-1 cells and isolated islets treated without or with 2 mM CLA. It is concluded that CLA may have a beneficial effect for maintenance of cell viability via increased Bcl-2 protein expression compared with steraic aced, which lead to delay or prevent, at least somewhat, the onset of NIDDM in the physiological status.

Effects of Vitamin C or E on the Pro-inflammatory Cytokines, Heat Shock Protein 70 and Antioxidant Status in Broiler Chicks under Summer Conditions

Jang, In-Surk,Ko, Young-Hyun,Moon, Yang-Soo,Sohn, Sea-Hwan Asian Australasian Association of Animal Productio 2014 Animal Bioscience Vol.27 No.5

The present study was carried out to investigate the effects of dietary antioxidants on pro-inflammatory cytokines, heat shock protein (HSP) and antioxidant status in broiler chicks under summer conditions. A total of 162, 3-d-old broiler chicks were randomly assigned to a basal diet (CON) and the basal diet supplemented with vitamin C (200 mg/kg diet, VCD) or vitamin E (100 mg/kg, VED) until 35 day of age. All birds were exposed to summer diurnal heat stress at average daily fluctuations of temperature between $32^{\circ}C$ to $34^{\circ}C$ at day to $27^{\circ}C$ to $29^{\circ}C$ at night for the entire feeding periods. There was no significant difference in body weight, feed to gain ratio and the relative organ weight except the thymus in response to dietary vitamin C or E supplementation. However, the mRNA expression of interleukin (IL)-$1{\beta}$, IL-6, interferon (IFN)-${\gamma}$, Toll like receptor (TLR)-4 and HSP70 in the liver of birds fed diet containing vitamin C significantly (p<0.05) decreased compared with those in birds fed basal diet. Dietary vitamin E also showed a significant (p<0.05) decrease in the mRNA expression of IL-6 and HSP70 compared with a basal diet. Total antioxidant status (TAS) in serum of birds fed vitamin C supplemented diet was significantly (p<0.05) higher with than that in birds a basal diet. Lipid peroxidation in serum and liver resulted in a significant (p<0.05) decrease in response to dietary vitamin C or E supplementation. In conclusion, dietary supplementation with antioxidant vitamins, especially vitamin C resulted in a significant decrease in the mRNA expression of pro-inflammatory cytokines and HSP70, and higher antioxidant parameters than that of birds on the basal diet under summer conditions.

RESEARCH : Effects of a lipid-encapsulated zinc oxide supplement on growth performance and intestinal morphology and digestive enzyme activities in weanling pigs

( In Surk Jang ),( Chang Hoon Kwon ),( Duck Min Ha ),( Dae Yun Jung ),( Sun Young Kang ),( Man Jong Park ),( Jeong Hee Han ),( Byung-chul Park ),( Chul Young Lee ) 한국동물자원과학회(구 한국축산학회) 2014 한국축산학회지 Vol.56 No.29

This study compared the effects of varying lipid content and dietary concentration of a lipid-encapsulated (LE) ZnO product to those of native ZnO and thereby to find insights into optimal lipid coating and dosage of the Zn supplement. A total of 192 21-d-old weanling pigs were allotted to 48 pens, after which each six pens received a ZnO-free basal diet supplemented with 125 ppm ZnO (100 ppm Zn; BASAL), 2,500 ppm Zn as native ZnO (HIGH), or 100 or 200 ppm Zn as LE ZnO (LE-100 or LE-250) containing 8%, 10%, or 12% lipid [LE-8%, LE-10%, or LE-12%, respectively; 2 × 3 factorial arrangement within the LE-ZnO diets (LE-ALL)] for 14 d. Forty pigs were killed at the end for histological and biochemical examinations. None of ADG, ADFI, gain:feed, and fecal consistency score differed between the LE-ALL and either of the BASAL and HIGH groups. Hepatic and serum Zn concentrations were greater (p <0.05) in the HIGH vs. LE-ALL group, but did not differ between LE-ALL and BASAL, between LE-100 and -250, or among LE-8%, -10%, and -12% groups. Villus height (VH), crypt depth (CD), and the VH:CD ratio in the duodenum, jejunum, and ileum did not differ between the LE-ALL and either of the BASAL and HIGH groups, except for a greater CD in the duodenum in the LE-ALL vs. HIGH group. Additionally, VH and CD in the duodenum and VH:CD in the jejunum were greater in the LE-250 vs. LE-100 group. Specific activities of sucrase, maltase, and leucine aminopeptidase in these intestinal regions and those of amylase and trypsin in the pancreas were not influenced by the lipid content or dietary concentration of LE ZnO and also did not differ between the LE-ALL and either of the BASAL and HIGH groups, except for a greater pancreatic amylase activity in the former vs. HIGH group. In conclusion, the present results indicate that the LE ZnO, regardless of its lipid percentage or supplementation level examined in this study, has no significant effect on growth performance, fecal consistency, or digestive enzyme activities of weanling pigs under the experimental conditions.

닭에서 사육밀도 및 Lipopolysaccharide 투여가 면역장기 무게, 혈액 생화학적 성상 및 친염증 사이토카인 mRNA 발현에 미치는 영향

장인석(In-Surk Jang),송민혜(Min-Hye Song),김하나(Ha-Na Kim),문양수(Yang Soo Moon),손시환(Sea Hwan Sohn) 한국가금학회 2016 韓國家禽學會誌 Vol.43 No.3

본 연구는 닭에서 사육밀도[(대조군(SSD) 495 ㎠/수 vs. 고밀도사육군(HSD) 245 ㎠/수] 및 LPS 투여(5 mg/kg BW)가 다른 각종 생리적 스트레스 지표에 미치는 영향을 조사하기 위해 실시되었다. 대조군, HSD군 및 LPS군(5 mg/kg BW)에서 체중 및 장기 무게를 조사한 결과, SSD군과 HSD군간 비교 시 체중은 고밀도사육 시 현저히(p<0.05) 감소하였으나, 간, 비장 및 흉선 무게는 차이가 없었다. LPS 투여군에서는 대조군에 비해 흉선과 F-낭 무게가 유의적(p<0.05)으로 증가하였다. 혈액 생화학적 성분에서 AST, ALT 및 BUN 수준은 SSD군과 HSD군 간에는 차이가 없었지만, LPS군에서 증가되었다(p<0.05). 혈중 Ca, P, CK, uric acid 수준 모두 LPS군에서유의하게(p<0.05) 높았다. 혈중 corticosterone 수준은 LPS군에서 SSD군과 HSD군에 비해 유의적(p<0.05)으로 증가되었으나, SSD와 HSD군 간에는 차이가 없었다. 친염증 사이토카인 mRNA 발현을 조사한 결과, 간 조직의 IL-1β, IL-6 및 iNOS mRNA 발현은 LPS군에서 SSD군에 비해 증가되었으나(p<0.05), SSD군과 HSD군 간에는 차이가 없었다. 흉선 IL-1β, IL-6, IL-18 mRNA 유전자 역시 LPS군에서 다른 군들에 비해 유의하게(p<0.05) 증가되었다. HSD군에서 흉선의 IL-1β는 SSD군에 비해 현저히(p<0.05) 증가되었다. F-낭의 IL-1β 발현은 LPS군에서 SSD군에 비해 증가되었으나(p<0.05), 다른 사이토카인은 차이가 없었다. 이상의 결과, 고밀도사육에 따른 만성스트레스 요인은 닭의 환경적응성에 따라 생산성을 제외한 다른 지표들의 변화는 적었으나, LPS 투여는 혈액 생화학성분 및 친염증 사이토카인 mRNA 발현을 현저하게 변화시켰다. This study was performed to investigate the effects of the stocking density (standard stocking density (SSD, 495 ㎠/bird)) vs. high stocking density (HSD,245㎠/bird) and challenge with lipopolysaccharide (LPS, 5mg/kg BW) on the stress-related physiological indicators in chicks. There was a significant (p<0.05) decrease in body weight, but not in the weight of immune organs, between the SSD and HSD groups. The LPS group resulted in a significant (p<0.05) increase in the weights of the thymus and bursa of fabricius compared with the SSD group. Plasma biochemical components, including aspartate transaminase (AST), alanine transaminase (ALT), blood urea nitrogen, Ca, P, creatine kinase and uric acid, markedly (p<0.05) increased in the LPS birds, although no difference in these parameters was observed between the SSD and HSD birds. Furthermore, the birds challenged with LPS showed a significant (p<0.05) increase in the plasma corticosterone level, although this hormone did not differ between the SSD and HSD groups. In the mRNA expression of pro-inflammatory cytokines, hepatic IL-1β, IL-6 and iNOS in the LPS group significantly (p<0.05) increased compared with those in the SSD group. Thymic mRNA expression of IL-1β, IL-6 and IL-18 in the LPS group also significantly (p<0.05) increased compared with those in the other groups. In addition, mRNA expression of IL-1β in the bursa of fabricius of the LPS group increased (p<0.05) without affecting the other cytokines. Under high stocking density, thymic IL-1β was the only cytokine that was up-regulated compared with the SSD group. In conclusion, an acute stress induced by LPS challenge profoundly affected immune organ weight, blood biochemical profiles and pro-inflammatory cytokine expression, while chronic stress did not markedly affect biochemical and immunological parameters, suggesting that chicks under high stocking density could be adapted to prolonged stressors.

질환모델동물에 있어서 생리적 특성의 발현 연구(Ⅰ) : 비만 모델 동물의 생리적 형질 발현 Physiological pattern of dbesity in potential obese laboratory animals

장인석,채갑용,강태석,김용규,김철규,황진희,황대연,이주은,허영범,정근기,조정식 식품의약품안전청 1998 식품의약품안전청 연보 Vol.2 No.-

수컷 SD 랫드를 사용하여, 핀탄수화물 급여군(HCHO), 탄수화물 및 지방 혼합 급여군(MPAT) 및 고지밟 급여군 (틴FAT), 그리고 절식군(R료I:T) 깊군을 체중웨 따란 완전임의 배치한 후 정제사료를 13주 동안 자옳 급식토록 하였다. 시험결과 체중에서는 절식관인 REST군을 제외하고 운의적 차이가 나타난지 않았으나, 사료 섭취량에서는 고수준의 탄순화물 급여군(HCHO)이 고지방 급여군(딘FAT)에 비해 유의적(P<0.95)으로 증가하였다. 혈중 glucose 함량은 HFAT군에서 다른 군에 비해 유의적으로 증가되었으나,중성지항은 오히려 고탄수왔물 급여군인 HCHO군엑서 증가하는 경향이 뚜렷하였다. 혈장내 insulin 농도는 HFAT군에서 현저한 증가를 보여 insvliu resistauce 현상을 나타내었다. 혈중 leptiBl농고는 MFAT와 HFAT군에서 HCHO와 REST군에 비해 유의적으로 증가하였으며, 특힉 절식군인 REST군에서는 HFAT와 HFAT군들에 비왜 약 3배 이상의 감소치를 보였다. Leptin receptor(extra-cellular와 intracellular donlain)의 mnNA 발현을 조사한 결과 영양소원 및 절식에 따른 영향 즉 혈중 leptin 함량에 따라서 leptin receptor왁 발현은 아문런 차이를 나타내지 않았다. NPY를 시상하부 3곳의 주요 nuclei(arcu-ate, paraventricular, supraopti매에서 면역 염잭하여 관찰한 결과 역시 혈중 leptin 및 insulin 함량파는 상관없이 HC려0, MfAT 및 HFAT군에서 모두 비슷한 양상을 보였다. 그러나 절식군인 REST군에서는 NPY 발현이 감소 하였다. 결론적으로 주에너지원으로 고지방 급여시 비만 및 당뇨가 유발되었으며, 증가된 혈중 leptin 맞 insulin은 41상31부 lertinreceptor 및 NPY조절을 통한 비만 조절 기전에 유의한 영향을 미치지 않았다. Leptin, prodv.cts of the adipose-specific oS gene, acts an important effect in the rrgulation of obesity. Several studies with obesity model animals have suggested that the action of leptin or-erted in the hypothalamus of the brain. However, in contrast to obesity model animals such as oo/oc, dD/08 mice aBd zucker rats,,dietary induced obesity model has not been intensively investigated on leptin'ssignaling pathway. Therefore, six wk old SD rats(n= 90) were allotted to HCHO(high carbohydratediet) group fed the diet couts.ming 70% carbohydrate and 11% fat, Mf'AT (medium fat diet) group frdthe diet containing 48% carbr)hydrate and 32.5% fat, HFAT(high fat diet) group fed the containing 33.3% carbohydrate and 42.8% fat, as basis of energy, and REST(restricted feeding) group that fed 50%restricted amount of ad fiSifHu3 of MFAT diet. All diets except REST consist of the salne energy densityand SB rats tvere continualty fed the purified diets for 13 wks. The body weight was not altered by die-tary treatrnftts eBcept REST group. However, daily feed intake increase4 in HCHO and MFAT grouprcompared with HFAT. Also, feeding HFAT enhanced serum glucose, insulin, and leptin levels when com-pared with IfCHO and REST groups. A second. stage was to determine whether the expressions of leptinreceptor and neuropeptide Y(Iffy ) in the hypothalamos were affected by serum leptin and insulia con-centrations. No difference in leptin receptor mRNA expression by RT-PCR were found amoBg groups. Inaddition, serum leptin and insulin did not alter NPY expression levels in arcuate, paraventricular, andsupraoptic nuclei in the hypothalamus. In summary, the expressions of leptin receptor and NPY arealtered by neither diet, nor leptin and iusulin levels in dietary induced obese SD rats.

Effects of Steaming Time and Frequency for Manufactured Red Liriope platyphylla on the Insulin Secretion Ability and Insulin Receptor Signaling Pathway

Sun Il Choi,Hye Ryun Lee,Jun Seo Goo,Ji Eun Kim,So Hee Nam,In Sik Hwang,Young Ju Lee,So Hae Prak,Hee Seob Lee,Jong Sup Lee,In Surk Jang,Hong Ju Son,Dae Youn Hwang 한국실험동물학회 2011 Laboratory Animal Research Vol.27 No.2

In oriental medicine, Liriope platyphylla (LP) has long been regarded as a curative herb useful for the treatment of diabetes, asthma, and neurodegenerative disorders. The principal objective of this study was to assess the effects of steaming time and frequency for manufactured Red LP (RLP) on insulin secretion ability and insulin receptor signaling pathway. To achieve our goal, several types of LPs manufactured under different conditions were applied to INS cells and streptozotocin (STZ)-induced diabetic ICR mice, after which alterations in insulin concentrations were detected in the culture supernatants and sera. The optimal concentration for the investigation of insulin secretion ability was found to be 50 ㎍/mL of LP. At this concentration, maximum insulin secretion was observed in the INS cells treated with LP extract steamed for 3 h (3-SLP) with two repeated steps (3 h steaming and 24 h air-dried) carried out 9 times (9-SALP); no significant changes in viability were detected in any of the treated cells. Additionally, the expression and phosphorylation levels of most components in the insulin receptor signaling pathway were increased significantly in the majority of cells treated with steaming-processed LP as compared to the cells treated with LP prepared without steaming. With regard to glucose transporter (GLUT) expression, alterations of steaming time induced similar responses on the expression levels of GLUT-2 and GLUT-3. However, differences in steaming frequency were also shown to induce dose-dependent responses in the expression level of GLUT-2 only; no significant differences in GLUT-3 expression were detected under these conditions. Furthermore, these responses observed in vitro were similarly detected in STZ-induced diabetic mice. 24-SLP and 9-SALP treatment applied for 14 days induced the down-regulation of glucose concentration and upregulation of insulin concentration. Therefore, these results indicated that the steaming processed LP may contribute to the relief of diabetes symptoms and should be regarded as an excellent candidate for a diabetes treatment.

Coenzyme Q10 첨가 급여가 산란계의 지방대사 연관 유전자 발현에 미치는 영향

장인석(In Surk Jang),문양수(Yang Soo Moon) 한국가금학회 2016 韓國家禽學會誌 Vol.43 No.1

Coenzyme Q10(CoQ10)은 자연계에 널리 분포하는 화합물로 세포호흡과 항산화제로서 그 기능이 잘 알려졌지만, 최근 유전자들의 발현 조절자로서의 가능성도 제시되었다. 따라서 본 연구는 산란계에서 CoQ10의 첨가 급이가 콜레스테롤과 지방산 대사관련 유전자들의 발현에 미치는 영향을 관찰하고자 실시하였다. Lohmann Brown(40주령) 36수를 CoQ-10의 첨가원에 따라 대조군(CON, basal diet(BD)), CoQ10 건조분말 급여군(T1, BD+CoQ10 100 ㎎/㎏ 사료) 및 CoQ10 건조분말 유화처리군(T2, BD+micellar of CoQ10 100 ㎎/㎏ 사료) 등 모두 3처리구로 설정하여 5주간 사양시험을 실시하였다. 시험 종료 후 각 개체의 간으로부터 total RNA를 추출하고, real-time PCR을 이용하여 유전자들의 발현을 분석하였다. 콜레스테롤 합성 과정에서 주요 조절 효소인 HMGCoA reductase(HMGCR)의 유전자 발현은 대조구에 비하여 CoQ10 분말첨가인 T1과 유화처리된 T2 처리구에서 모두 약 50%씩 억제되었다(p<0.05). 내생 콜레스테롤의 합성을 촉진시키는 전사인자인 SREBP2 mRNA 발현 또한 대조구와 비교해서 T1과 T2에서 각각 30%와 40% 감소하였다(p<0.05). CoQ10의 첨가 급이는 대조구에 비하여 liver X receptor(LXR) 유전자가 약 30∼35% 그 발현이 억제되었으며, sterol regulatory element-binding proteins(SREBPs)1 또한 T2에서 약 40% 유전자 발현이 감소하였다(P<0.05). 전사인자인 PPARγ와 XBP1은 CoQ10에 의하여 약 15∼40% 수준으로 효과적으로 억제됨을 확인하였다(p<0.05). 세포 내부로의 에너지 공급원인 포도당의 흡수를 담당하는 GLUT2는 약 35∼60% 그리고 GLUT8은 약 25∼30%의 유전자발현 각각 감소함을 보였다(p<0.05). CoQ10의 섭취는 중성지방 합성을 위한 지방합성효소(FASN)의 유전자 발현을 분말처리군에서 약 30%, 유화처리군에서 약 65% 억제됨을 확인하였다(P<0.05). 본 연구결과는 CoQ10 첨가급여가 콜레스테롤 및 지방대사 관련 유전자 발현에 영향을 미치며, 세포내 콜레스테롤과 지방의 생성도 억제할 수 있음을 보여주었다. The aim of this study was to investigate the expression patterns of key genes involved in lipid metabolism in response to dietary Coenzyme Q10 (CoQ10) in hens. A total of 36 forty week-old Lohmann Brown were randomly allocated into 3 groups consisting of 4 replicates of 3 birds. Laying hens were subjected to one of following treatments: Control (BD, basal diet), T1 (BD+ CoQ10 100 ㎎/㎏ diet) and T2 (BD+ micellar of CoQ10 100 ㎎/㎏ diet). Birds were fed ad libitum a basal diet or the basal diet supplemented with CoQ10 for 5 weeks. Total RNA was extracted from the liver for quantitative RT-PCR. The mRNA levels of HMG-CoA reductase(HMGCR) and sterol regulatory element-binding proteins(SREBP)2 were decreased more than 30∼50% in the liver of birds fed a basal diet supplemented with CoQ10 (p<0.05). These findings suggest that dietary CoQ10 can reduce cholesterol levels by the suppression of the hepatic HMGCR and SREBP2 genes. The gene expressions of liver X receptor (LXR) and SREBP1 were down regulated due to the addition of CoQ10 to the feed (p<0.05). The homeostasis of cholesterol can be regulated by LXR and SREBP1 in cholesterol-low-conditions. The supplement of CoQ10 caused a decreased expression of lipid metabolism-related genes including PPARγ, XBP1, FASN, and GLUTs in the liver of birds (p<0.05). These data suggest that CoQ10 might be used as a dietary supplement to reduce cholesterol levels and to regulate lipid homeostasis in laying hens.

Lipopolysaccride 감염처리가 닭의 품종간 스트레스연관 유전자 발현에 미치는 영향

장인석(In Surk Jang),손시환(Sea Hwan Sohn),문양수(Yang Soo Moon) 한국가금학회 2017 韓國家禽學會誌 Vol.44 No.1

본 연구는 한국재래계(KNC)와 백색레그혼(WLH)에서 lipopolysaccharide(LPS) 감염 스트레스가 닭의 품종간 스트레스 연관 유전자들의 발현에 미치는 영향을 비교․분석하고자 실시되었다. 공시계를 대상으로 생리식염수(대조구)와 LPS(처리구)를 복강에 투여한 후, 시간(0, 48 hr) 및 처리별 각 개체로부터 간 조직을 취하고, microarray 및 quantitative RT-PCR(qRT-PCR) 분석을 하였다. 처리에 따른 유전자 발현차이를 보면, KNC(대조구)와 KNC에 LPS를 처리한 경우(KNC-LPS)를 비교한 결과, 대조구 대비 2배 이상 유전자의 발현이 증가한 유전자의 수는 1,044개, 발현이 감소한 유전자의 수는 1,000개였다. WLH(대조구)를 WLH-LPS와 비교한 경우, 유전자의 발현이 증가한 유전자의 수는 1,193개, 발현이 감소한 유전자의 수는 1,072개였다. LPS 처리에 따른 스트레스 연관 유전자들의 microarray 발현에서 스트레스연관 유전자들의 발현은 두 품종 모두에서 감소하였으며, 품종 간 차이는 없는 것으로 나타났다. Microarray의 결과를 바탕으로 HSP90, HMGCR, ATF4, SREBP1, XBP1 등의 유전자 발현을 qRT-PCR을 이용하여 검증한 결과, 대조구와 LPS 감염구 간에 유의적 차이를 나타내었다(P<0.05). 세포수준의 스트레스(ER 스트레스)에서 ATF4, XBP1, SREBP1은 화이트레그혼에서 microarray와 qRT-PCR에서와 같이 이들 유전자들의 발현이 억제되는 것을 보여주었다. 그러나 한국재래계에서는 ATF4를 제외한 유전자들은 LPS에 의해 영향을 받지 않거나(XBP1), 오히려 증가(SREBP)하는 양상을 보였다. ER-stress 연관 유전자들의 발현 양상으로 볼 때, KNC이 WLH에 비하여 LPS 감염에 더 민감하게 반응하는 것으로 보인다. HMGCR은 두 품종간에 LPS에 의한 상호작용이 없는 것으로 보아, HMGCR 발현에 의한 감염 차이점을 찾을 수 없었다. 한국재래계에서 HSP70은 LPS 처리 후에 대조구에 비하여 약 2.5배 이상 높은 발현을 보였으나, 백색 레그혼에서는 낮은 발현 양상을 나타내었다. 스트레스지표 유전자들의 종류뿐만 아니라, 스트레스 종류(예: 환경스트레스, 감염스트레스)에 따라 유전자들의 발현 반응에 차이가 있음을 보여주었다. LPS 감염스트레스에 따른 스트레스연관 유전자 발현연구는 닭의 품종별 질병 저항성 및 동물복지 관련 지표의 탐색에 기여할 것으로 사료된다. The objective of the present study was to determine the expression of genes associated with lipopolysaccharide (LPS)-induced stressor in two breeds of chickens: the Korean native chicken (KNC) and the White Leghorn chicken (WLH). Forty chickens per breed, aged 40 weeks, were randomly allotted to the control (CON, administered the saline vehicle) and LPSinjected stress groups. Samples were collected at 0 and 48 h post-LPS injection, and total RNA was extracted from the chicken livers for RNA microarray and quantitative real-time polymerase chain reaction (qRT-PCR) analyses. In response to LPS, 1,044 and 1,193 genes were upregulated, and 1,000 and 1,072 genes were downregulated in the KNC and WLH, respectively, using a≥2-fold cutoff change. A functional network analysis revealed that stress-related genes were downregulated in both KNC and WLH after LPS infection. The results obtained from the qRT-PCR analysis of mRNA expression of heat shock 90 (HSP90), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), activating transcription factor 4 (ATF4), sterol regulatory element-binding protein 1 (SREBP1), and X-box binding protein 1 (XBP1) were confirmed by the results of the microarray analysis. There was asignificant difference in the expression of stress-associated genes between the control and LPS-injected KNC and WLH groups. The qRT-PCR analysis revealed that the stress-related HSP90α and HMGCR genes were downregulated in both LPS-injected KNC and WLH groups. However, the HSP70 and HSP90β genes were upregulated only in the LPS-injected KNC group. The results suggest that the mRNA expression of stress-related genes is differentially affected by LPS stimulation, and some of the responses varied with the chicken breed. A better understanding of the LPS-induced infective stressors in chicken using the qRTPCR and RNA microarray analyses may contribute to improving animal welfare and husbandry practices.

한방생약부산물 급여가 육계의 생산성, 장관 미생물 균총, 혈액 생화학적 성상 및 면역학적 지표에 미치는 영향

장인석(In-Surk Jang) 한국가금학회 2015 韓國家禽學會誌 Vol.42 No.4

본 연구는 한방생약부산물을 이용한 육계의 사료첨가제를 개발하기 위해 대조군(CON), HM1(한방생약부산물 0.15%), HM2(한방생약부산물 0.3%) 및 HM3(한방생약부산물 0.5%) 등 4개군에 육계 96수를 완전임의 배치하여 생산성, 장내 미생물 균총, 혈액 생화학적 지표, 면역글로블린 및 백혈구 성상의 변화에 대하여 조사하였다. 한방생약부산물 0.15% 첨가군(HM1)에서 대조군에 비해 육계전기(3∼21일령) 사료요구율이 유의적(p<0.05)으로 개선되었으나, 전 기간에서는 차이가 없었다. 장내 소화물의 E. coli 및 Lactobacili 총 균수에서도 한방생약부산물 급여는 영향을 미치지 않았다. 혈액 생화학적 지표에서는 한방생약부산물 급여 시 AST 및 glucose 수준을 유의하게(p<0.05) 감소시켰다. 면역글로불린(IgG 및 IgA)과 백혈구 감별계수에서는 처리에 따른 차이는 없었다. 결론적으로 육계에서 0.15∼0.3% 수준의 한방생약 부산물 첨가는 육계전기에 사료이용성 및 간 조직의 기능을 개선하는 효과가 있는 것으로 사료된다. The study was carried out to investigate the effects of by-products of herbal medicines on performance, enteric microflora and blood biochemical profiles and immunological parameters in broiler chicks. A total of ninety-six, 3-d-old birds were assigned to a basal diet (CON) or a basal diet supplemented with 0.15 % (HM1), 0.3% (HM2) or 0.5% (HM3) by-products of herbal medicines. There was a significantly (p<0.05) improved feed conversion ratio (FCR) in birds fed diet supplemented with 0.15% by-products of herbal medicines (HM1) compared with the control birds during starter period (3∼21 days). However, no difference in body weight, feed intake, total gain and FCR among treatment groups was observed during the entire feeding period (3∼35 days). The colony forming units (CFU) of E. coli and Lactobacilli in the digesta of ileo-cecum was not also affected by dietary treatment. Serum AST (aspartate aminotransferase) and glucose decreased (p<0.05) in birds fed diets supplemented with herbal medicines compared with those fed the basal diet. In particular, the birds fed diets supplemented with herbal medicines showed a significant (p<0.05) increase in blood mean corpuscular volume (MCV) level compared with the control birds. However, the most of blood biochemical and hematological parameters and immunoglobulins (IgG and IgA) were not affected by the dietary treatment. In conclusion, the low level of dietary herbal medicines showed beneficial effects on FCR during starter period and liver functions, suggesting that by-products of herbal medicines may be applicable as functional feed additives in birds.

육계에서 가시오갈피 급여에 따른 생산성, 혈액 생화학적 성상 및 면역 사이토카인 발현에 미치는 영향

장인석,문양수,손시환,Jang, In-Surk,Moon, Yang Soo,Sohn, Sea Hwan 한국가금학회 2015 韓國家禽學會誌 Vol.42 No.3

본 연구는 천연항산화 생리활성 소재로서 가시오갈피를 육계에게 0(CON), 0.5(AS1) 및 1.0%(AS2) 수준으로 급여하여 생산성, 면역장기 무게, 혈액 생화학적 성상 및 친염증 사이토카인 mRNA 발현에 미치는 영향을 조사하였다. 전 사육기간에서 가시오갈피 급여에 따른 체중, 증체 및 사료섭취량은 유의적 차이를 보이지 않았으나, AS2구에서 사료요구율이 유의적으로(p<0.05) 저하되었다. 면역장기 무게를 관찰한 결과, 간, 비장, F-낭 및 흉선 무게는 가시오갈피 급여에 따른 영향이 없었다. 혈액 생화학적 성분에서 albumin, total protein, cholesterol, AST, ALT 등과 같은 대부분의 성분은 대조구와 처리구간 차이는 없었다. 그러나 triglycerides는 대조구에 비해 AS2구에서 현저히(p<0.05) 높았고, glucose 수준은 대조구에 비해 가시오갈피 급여구에서 현저히(p<0.05) 증가되었다. 친염증 사이토카인 mRNA 발현을 조사한 결과, 가시오갈피 급여에 따라 백혈구의 IFN-${\gamma}$ mRNA 발현이 현저히(p<0.05) 감소되었으며, IL1-${\beta}$, IL-6, TNF-${\alpha}$, iNOS 등은 유의적 차이는 없었지만, 감소되는 경향이 있었다. 비장에서 친염증 사이토카인 발현은 유의적 차이가 없었다. 간 조직에서는 0.5% 가시오갈피 급여에 따라 iNOS mRNA 발현이 유의적으로(p<0.05) 감소하였지만, 다른 친염증 사이토카인 발현은 유의적 차이는 없었다. 따라서 육계 생산성 및 친염증 사이토카인 발현에 미치는 영향 등을 고려할 경우, 0.5% 수준의 가시오갈피 급여가 가장 바람직한 것으로 판단된다. This study was performed to examine the effects of dietary Acanthopanax senticosus (AS) on growth performance, immune organ weights, blood biochemical parameters and the expression of pro-inflammatory cytokines in broiler chicks. A total of 120 4-day-old birds were given a basal diet (CON) or a basal diet supplemented with 0.5% (AS1) or 1.0% (AS2) AS powder until the birds were 35 days of age. There was no difference in body weight, total gain, feed intake or immune organ weights among the treatment groups. However, the feed conversion ratio in the AS2 group was lower (p<0.05) than that in the CON group. Serum biochemical components, including AST (aspartate aminotransferase), ALT (alanine aminotransferase), albumin and total protein, were not affected by the dietary treatments, whereas glucose and triglyceride levels increased (p<0.05) in the AS2 group compared with the CON group. The AS1 group exhibited decreased mRNA expression (p<0.05) of IFN-${\gamma}$ in white blood cells and iNOS in the liver compared with the CON group. The other pro-inflammatory cytokines were unaffected by dietary AS supplementation, although there was a trend towards decreased expression of these genes, including those encoding Il-$1{\beta}$, IL-6 and TNF-${\alpha}$. In conclusion, dietary supplementation with 0.5% AS decreased the expression of several pro-inflammatory cytokines without affecting growth performance, suggesting that this supplement might be applicable as an immunoregulatory feed additive in broiler chicks.