Glucagon 검사시 Aprotinin 첨가와 Plastic tube 사용이 미치는 영향

조윤교,최삼규,서소연,신용환,Cho, Youn-Kyo,Choi, Sam-Kyu,Seo, So-Yeon,Shin, Yong-Hwan 대한핵의학기술학회 2011 핵의학 기술 Vol.15 No.1

Glucagon은 췌장 Langerhans섬 ${\alpha}$-세포에서 합성 분비되며, 기능으로는 간 당원분해, 지방분해, 인슐린분비 촉진 작용 등이 있다. Glucagon의 측정은 Glucagon 과잉증에 의한 당뇨병, 특발성 Glucagon 결손증, 불안정 당뇨병에서 저혈당의 진단을 하기위해 실시된다. Glucagon 측정시 세 가지 주의를 요하는데, 첫째, Aprotinin이 첨가된 EDTA tube에 채혈해야 하고 둘째, Plasma를 분리하여 Glass tube에 냉동 보관하여야 하고, 마지막으로 검사시에도 Glass tube에서 측정을 해야 한다. 이에 우리는 EDTA tube에 Aprotinin의 유무에 따른 결과 비교, 검체 보관 용기의 차이에 따른 비교(Plastic tube/ Glass tube), 측정용기에 따른 비교(Plastic tube/Glass tube)를 하여, 세 가지가 결과에 미치는 영향을 알아보고자 한다. 성별 구분 없이 건강검진에서 정상 소견을 보인 성인 40명을 대상으로 실험하여 대조군과 세가지 다른 실험군의 Glucagon 결과를 비교하였다. Aprotinin첨가한 EDTA 용기에 채혈 후, 검체를 Glass tube에 3일 냉동 보관하여 Glass tube에서 실험(대조군)한 결과와 Aprotinin첨가하지 않은 EDTA 용기에 채혈한 결과(실험군1), Plastic tube에 3일 냉동 보관한 후의 결과(실험군2), Plastic tube에서 실험(실험군3)한 결과를 비교하였다. 통계적인 분석은 SPSS를 이용한 paired t-test와 단순선형회귀분석을 실시하였고 시약은 상품화된 Siemens사의 Glucagon RIA kit를 사용하였다. Aprotinin이 첨가된 EDTA 용기에 채혈하여 실험한 결과와 Aprotinin이 첨가되지 않은 EDTA용기에 채혈하여 실험한 상관계수는 r=0.783(p=0.064)를 보였다. 검체 분리 후 Plastic tube에서 3일 냉동 보관하여 실험한 결과는 Glass tube의 결과에 비해서 유의하게 낮게 나타났다(r=0.979, p=0.005). 또 측정 시 Plastic tube를 사용한 결과도 Glass tube에서 실험한 결과보다 유의하게 낮게 나타났다(r=0.754, p<0.001). Glucagon 검사 시에는 단백 분해효소 억제제인 Aprotinin이 첨가된 EDTA tube 사용이 권장되며, Plastic tube 사용 시 환자의 결과가 유의하게 낮게 나타나므로 검체는 분리 후 Glass tube에 냉동 보관하고 Glass tube에서 실험하여야 한다. Purpose: There are 3 warnings for Glucagon tests. First, EDTA tubes that already contain Aprotinin must be used for plasma collection. Second, for freezer storage of centrifuged plasma, glass tubes must be used. Last, glass tubes must be used for testing procedure. So we compared the glucagon results of next 3 situation to those of control group. First, We compared to results by tubes without Aprotinin and with aprotinin. Second, we compared to results by tubes(plastic vs glass) for plasma storage. Third, we compared to results by tubes(plastic vs glass) for testing. We tried to evaluate the results of the 3 different condition. Materials and Methods: 40 healthy adults were studied with normal results on the general medical check up and laboratory tests. We compared the results of 3 different condition belows: Blood were collected in EDTA tube containing aprotinin and plasma was stored in the glass tube for 3 days in a freezer and results were obtained by tests in the glass tubes. Results from EDTA plasma without aprotinin, results from platic tubes for freezer stroage, results from plastic tube when testing. Simple linear regression analysis and paired t-test using SPSS were done for statistical analysis. Commercial glucagon kit(RIA-method)which made by Siemens company were used. Results: Correlation coefficient between results of EDTA tubes with Aprotinin vs without Aprotinin was r=0.783 (p=0.064). Result of specimen in plastic tubes stored 3 days in a freezer showed lower value compared to those in glass tube(r=0.979, p=0.005). Also, results of testing in plastic tubes showed lower values than those testing in glass tubes. (r=0.754, p<0.001). Conclusion: It is recommended for glucagon determination to use EDTA tube with Aprotinin which is a inhibitor of protein breakdown enzyme. Results of plastic tube when storage and testing showed lower value than those of glass tubes, so it is recommended to store and test in glass tubes.

Glucagon과 insulin이 glutathione 항상성에 미치는 영향: 세포신호전달체계 및 glutathione transport system의 역할

김봉희,오정민,윤강욱,김충현,김상겸,Kim, Bong-Hee,Oh, Jung-Min,Yun, Kang-Uk,Kim, Chung-Hyeon,Kim, Sang-Kyum 환경독성보건학회 2007 환경독성보건학회지 Vol.22 No.3

It has been reported that hepatic glutathione (GSH) levels are decreased in diabetic patients, and glucagon increases hepatic efflux of GSH into blood. The signaling pathways responsible for mediating the glucagon effects on GSH efflux, however, are unknown. The signaling pathways involved in the regulation of GSH efflux in response to glucagon and insulin were examined in primary cultured rat hepatocytes. The GSH concentrations in the culture medium were markedly increased by the addition of glucagon, although cellular GSH levels are significantly decreased by glucagon. Insulin was also increased the GSH concentrations in the culture medium, but which is reflected in elevations of both cellular GSH and protein. Treatment of cells with 8-bromo-cAMP or dibutyryl-cAMP also resulted in elevation of the GSH concentrations in the culture medium. Pretreatment with H89, a selective inhibitor of protein kinase A, before glucagon addition markedly attenuated the glucagon effect. These results suggest that glucagon changes GSH homeostasis via elevation of GSH efflux, which may be responsible for decrease in hepatic GSH levels observed in diabetic condition. Furthermore, the present study implicates cAMP and protein kinase A in mediating the effect of glucagon on GSH efflux in primary cultured rat hepatocytes.

Design, synthesis, and effects of novel phenylpyrimidines as glucagon receptor antagonists

Choi, Hojung,Lee, Chang-Yong,Park, Eun-Young,Lee, Kyoung Mee,Shin, Dongyun,Jun, Hee-Sook Elsevier 2018 Bioorganic & medicinal chemistry Vol.26 No.21

<P><B>Abstract</B></P> <P>The hormone glucagon increases blood glucose levels through increasing hepatic glucose output. In diabetic patients, dysregulation of glucagon secretion contributes to hyperglycemia. Thus, the inhibition of glucagon receptor is one target for the treatment of hyperglycemia in type 2 diabetes. Here we designed and synthesized a series of small molecules based on phenylpyrimidine. Of these, the compound <B> <I>(R)</I>-7a</B> most significantly decreased the glucagon-induced cAMP production and glucagon-induced glucose production during <I>in vitro</I> and <I>in vivo</I> assays. In addition, <B> <I>(R)</I>-7a</B> showed good efficacy in glucagon challenge tests and lowered blood glucose levels in diabetic <I>db/db</I> mice. Our results suggest that the compound <B> <I>(R)</I>-7a</B> could be a potential glucose-lowering agent for treating type 2 diabetes.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Design and synthesis of novel GCGR antagonists as pyrimidine derivatives. </LI> <LI> The compound of <B> <I>(R)</I>-7a</B> decreased the glucagon-induced cAMP production. </LI> <LI> The compound of <B> <I>(R)</I>-7a</B> decreased the glucagon-induced glucose production. </LI> <LI> The compound of <B> <I>(R)</I>-7a</B> decreased the glucagon-induced glucose excursion <I>in vivo</I>. </LI> <LI> The compound of <B> <I>(R)</I>-7a</B> lowering fasting blood glucose levels in <I>db</I>/<I>db</I> mice. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

방사선이 마우스 위장관에 미치는 glucagon의 면역조직화학적 영향

김부순,조광호,박상옥 한국위생과학회 2002 한국위생과학회지 Vol.8 No.1

방사선 조사가 위장관 각 부위(위저부, 유문뷰, 십이지장, 공장, 회장, 맹장, 결장 및 직장)에 미치는 영향을 관찰하기 위하여 비교적 방사선에 민감한 strain으로 알려진 C57BL/6 마우스의 위장관 부위에 국소적으로 2 또는 4 Gy의 gamma 선을 조사하여, 8시간 후 위장관의 변화를 관찰하였다. 위장관에 존재하는 내분비세초의 혈 중 glucagon의 수치와 호르몬을 생산하는 내분비세포의 수적 및 분포의 변화를 각각의 항혈청을 이용한 면역조직화적 방법으로 위장관 각 부위에서 관찰하였던 바 다음과 같은 결과를 얻었다. Glucagon 면역반응세포는 정상군을 포함한 모든 실험군에서 위저부에 국한되어 관찰되었으며, 수적 변동은 관찰되지 않았으나, 방사선 조사선량에 비례하여 탈과립을 보이는 세포들의 수적 증가가 인정되었다. 한편 혈중 glucagon함량은 상층 조사군에서만 증가되었으며, 중간층 및 하층 조사군에서는 정상군과 유사한 수치를 나타내었다. 이상에서 방사선 조사 후 내분비세포의 감소는 면역조직화학적으로 염색되는 내분비세포의 과립이 혈 중에 유리된 결과로 생각되며, 다른 인자에 의한 탈과립의 자극보다는 방사선이 직접 면역반응세포에 작용할 것으로 생각되었다. To demonstrate the effect of local gamma irradiation on the gastrointestinal tract of the radiation sensitive C57BL/6 mice strain, the histological profiles of the 8 portions of gastrointestinal tract(fundus, pylorus, duodenum, jejunum, ileum, cecum, colon and rectum) were observed 8 hours after irradiation(subdivided into 3 parts, upper, middle and lower irradiated group). Gastrointestinal endocrine cells were determined by immunohistochemical methods using specific antisera against glucagon with their serum levels by radioimmynoassay(RIA). Glucagon-immunoreactive cells were restricted in the fundus of all tested groups including non-irradiated normal group, similar number of these immunoreactive cells were demonstrated in all tested groups. While increasing of cell numbers showing degranulation were found found dependent with amount of irradiated gamma ray. However, serum glucagon levels increased with amount of irradiated gamma ray in the case of upper irradiated group. In lower and middle irradiated group, no changes of serum glucagon levels were demonstrated in this study. In conclusion, Hormone in blood increased, well corresponding to those of decrease of immunoreactive hormone- producing cells in gastrointestinal tract.

Higher glucagon-to-insulin ratio is associated with elevated glycated hemoglobin levels in type 2 diabetes patients

이민영,김민경,박종숙,이상배,유지홍,안철우,김경래,강신애 대한내과학회 2019 The Korean Journal of Internal Medicine Vol.34 No.5

Background/Aims: The importance of α-cell dysfunction in the pathogenesis of type 2 diabetes has re-emerged recently. However, data on whether relative glucagon excess is present in clinical settings are scarce. We aimed to investigate associations between glucagon-to-insulin ratio and various metabolic parameters. Methods: A total of 451 patients with type 2 diabetes naïve to insulin treatment were recruited. Using glucagon-to-insulin ratio, we divided subjects into quartiles according to both fasting and postprandial glucagon-to-insulin ratios. Results: The mean age of the subjects was 58 years, with a mean body mass index of 25 kg/m2. The patients in the highest quartile of glucagon-to-insulin ratio had higher glycated hemoglobin (HbA1c) levels. HbA1c levels were positively correlated with both fasting and postprandial glucagon-to-insulin ratios. Subjects in the highest quartile of postprandial glucagon-to-insulin ratio were more likely to exhibit uncontrolled hyperglycemia, even after adjusting for confounding factors (odds ratio, 2.730; 95% confidence interval, 1.236 to 6.028; p for trend < 0.01). Conclusions: Hyperglucagonemia relative to insulin could contribute to uncontrolled hyperglycemia in type 2 diabetes patients.

Effects of 6-Month Sitagliptin Treatment on Insulin and Glucagon Responses in Korean Patients with Type 2 Diabetes Mellitus

양혜경,강보라미,이승환,김헌성,윤건호,차봉연,조재형 대한당뇨병학회 2015 Diabetes and Metabolism Journal Vol.39 No.4

Background: This study aimed to evaluate the effect of sitagliptin, an oral dipeptidyl peptidase-4 inhibitor, on insulin secretion and glucagon suppression in Korean subjects with type 2 diabetes mellitus. Methods: Twenty-four subjects underwent a 75-g oral glucose tolerance test (OGTT) before and after 6 months of sitagliptin treatment. Sitagliptin, insulin, and sulfonylurea were withdrawn for 3 days before OGTT to eliminate any acute effects on β-cell insulin or α-cell glucagon secretion. Venous samples were drawn five times during each OGTT to measure plasma glucose, insulin, and glucagon. Indices on insulin secretion and resistance were calculated. Results: Early phase insulin secretion, measured by the insulinogenic index significantly increased after 6 months of sitagliptin treatment, especially in the higher baseline body mass index group and higher baseline glycosylated hemoglobin (HbA1c) group. There were no significant differences in the insulin resistance indices before and after sitagliptin treatment. Although no significant differences were observed in the absolute levels of glucagon and the glucagon-to-insulin ratio, there was a significant reduction in the percentile change of glucagon-to-insulin ratio at 30- and 120-minute during the OGTT. Conclusion: Although the HbA1c level did not decrease significantly after 6 months of sitagliptin treatment, an increase in insulin secretion and reduction in early phase postprandial plasma glucagon-to-insulin ratio excursion was confirmed in Korean subjects with type 2 diabetes.

Glucagon-Like Peptide-1 Based Therapies: A New Horizon in Obesity Management

손장원,임수 대한내분비학회 2024 Endocrinology and metabolism Vol.39 No.2

Obesity is a significant risk factor for health issues like type 2 diabetes and cardiovascular disease. It often proves resistant to traditional lifestyle interventions, prompting a need for more precise therapeutic strategies. This has led to a focus on signaling pathways and neuroendocrine mechanisms to develop targeted obesity treatments. Recent developments in obesity management have been revolutionized by introducing novel glucagon-like peptide-1 (GLP-1) based drugs, such as semaglutide and tirzepatide. These drugs are part of an emerging class of nutrient-stimulated hormone-based therapeutics, acting as incretin mimetics to target G-protein–coupled receptors like GLP-1, glucose-dependent insulinotropic polypeptide (GIP), and glucagon. These receptors are vital in regulating body fat and energy balance. The development of multiagonists, including GLP-1–glucagon and GIP–GLP-1–glucagon receptor agonists, especially with the potential for glucagon receptor activation, marks a significant advancement in the field. This review covers the development and clinical efficacy of various GLP-1-based therapeutics, exploring the challenges and future directions in obesity management.

다발성 글루카곤 생성 내분비세포종을 동반한 췌장 알파세포의 Nesidioblastosis 및 과다증식 1예

강화평 ( Hua Pyong Kang ),김세화 ( Se Wha Kim ),임태섭 ( Tae Seop Lim ),이혜원 ( Hye Won Lee ),최흔 ( Heun Choi ),강창무 ( Chang Moo Kang ),김호근 ( Ho Guen Kim ),방승민 ( Seung Min Bang ) 대한소화기학회 2014 대한소화기학회지 Vol.63 No.4

Nesidioblastosis is a term used to describe pathologic overgrowth of pancreatic islet cells. It also means maldistribution of islet cells within the ductules of exocrine pancreas. Generally, nesidioblastosis occurs in beta-cell and causes neonatal hyperinsulinemic hypoglycemia or adult noninsulinoma pancreatogenous hypoglycemia syndrome. Alpha-cell nesidioblastosis and hyperplasia is an extremely rare disorder. It often accompanies glucagon-producing marco- and mircoadenoma without typical glucagonoma syndrome. A 35-year-old female was referred to our hospital with recurrent acute pancreatitis. On radiologic studies, 1.5 cm sized mass was noted in pancreas tail. Cytological evaluation with EUS-fine-needle aspiration suggested serous cystadenoma. She received distal pancreatectomy. The histologic examination revealed a 1.7 cm sized neuroendocrine tumor positive for immunohistochemical staining with glucagon antibody. Multiple glucagon-producing micro endocrine cell tumors were scattered next to the main tumor. Additionally, diffuse hyperplasia of pancreatic islets and ectopic proliferation of islet cells in centroacinar area, findings compatible to nesidioblastosis, were seen. These hyperplasia and almost all nesidioblastic cells were positive for glucagon immunochemistry. Even though serum glucagon level still remained higher than the reference value, she has been followed-up without any evidence of recurrence or hormone related symptoms. Herein, we report a case of alpha-cell nesidioblastosis and hyperplasia combined with glucagon-producing neuroendocrine tumor with literature review.

Pancreatic α-Cell Dysfunction in Type 2 Diabetes: Old Kids on the Block

문준성,원규장 대한당뇨병학회 2015 Diabetes and Metabolism Journal Vol.39 No.1

Type 2 diabetes (T2D) has been known as ‘bi-hormonal disorder’ since decades ago, the role of glucagon from α-cell has languished whereas β-cell taking center stage. Recently, numerous findings indicate that the defects of glucagon secretion get involve with development and exacerbation of hyperglycemia in T2D. Aberrant α-cell responses exhibit both fasting and postprandial states: hyperglucagonemia contributes to fasting hyperglycemia caused by inappropriate hepatic glucose production, and to postprandial hyperglycemia owing to blunted α-cell suppression. During hypoglycemia, insufficient counter-regulation response is also observed in advanced T2D. Though many debates still remained for exact mechanisms behind the dysregulation of α-cell in T2D, it is clear that the blockade of glucagon receptor or suppression of glucagon secretion from α-cell would be novel therapeutic targets for control of hyperglycemia. Whereas there have not been remarkable advances in developing new class of drugs, currently available glucagon-like peptide-1 and dipeptidyl peptidase-IV inhibitors could be options for treatment of hyperglucagonemia. In this review, we focus on α-cell dysfunction and therapeutic potentials of targeting α-cell in T2D.

Immunohistochemical studies of glucagon and somatostatin in the pancreas of the Korean tree squirrel. Sciurus vlugar is corea

이형식,이재현,Lee, Hyeung-sik,Lee, Jae-hyun The Korean Society of Veterinary Science 1993 大韓獸醫學會誌 Vol.33 No.4

청설모(Sciurus vulgaris corea)의 췌장에 분포하는 glucagon 및 somatostatin 세포의 분포와 그들 분비과립의 특징을 밝히기 위해 면역조직화학적 및 면역전자현미경적 방법으로 관찰하였다. Glucagon 면역반응세포는 주로 췌도의 주변부에 분포하였고, 때로는 외분비부에서 관찰되었다. 이 세포의 분비과립은 직경이 240~320nm였으며, 전자밀도가 높은 core를 전자밀도가 낮은 halo가 둘러싸고 특히 gold particle들은 전자밀도가 높은 core에 강한 반응을 보였다. Somatostatin면역반응세포는 췌도의 주변부를 따를 면역반응이 아주 약한 세포들로 산재하였으며 또한 외분비에서 단독 혹은 소집단으로 관찰되었다. 이 세포의 분비과립은 전자밀도가 낮고 직경이 250~275nm였으며, gold particle는 분비과립에 중등도로 고른 반응을 보였다. The pancreatic endocrine cells, glucagon and somatostation, of the Korean tree squirrel. Sciurus vulgais corea, were investigation by means of light and electron microscopic immunohistochemistry using the PAP and protein A-gold techniques. Glucagon-immunoreactive cells were distributed the periphery and occasinonaly central region of the pancreatic islets. Also, isolated cell was found between the pancreatic ancinar cells. The glucagon cells contraine granules with a diameter of 240~320nm and the electron dense core usually surrounded by a halo of less dense granular material. The core of granule was labelled strongly with gold particles. Somatostatin-immunoreactive cells were weakly stained in scattered along the peripheral pancreatic islets and were distributed as singly or small groups with in the pancreatic acinar cells. The somatostatin cells were spherical with a diameter of 250~275nm, moderately electron opaque (Gold particles were mostly demonstrated on the entire granule.