The objective of the present study was to evaluate the potential antidiabetic and antioxidant effect of the ethanol extract from Chrysanthemum cornarium L. var. spatiosum(CSE) against alloxan-induced oxidative stress in pancreatic β-cells, HIT-T15. In this study, the antidiabetic effect of CSE was examined using the 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazoliu bromide(MTT) cell proliferation assay, lactate dehydrogenase(LDH) release assay, NAD+/NADH ratio and insulin secretion. To further investigate whether CSE is involved in the antioxidant activity of alloxan-damaged HIT-T15 cells, its antioxidant effect against alloxan-induced oxidative stress was measured in HIT-T15 cells by determining the levels of antioxidant enzymes including superoxide dismutase(SOD), glutathione S-transferase(GST), glutathione reductase(GR) and glutathione peroxidase(GPx). The results of this analysis showed that alloxan significantly decreased cell viability, increased LDH leakage, and lowered NAD+/NADH ratio and insulin secretion in HIT-T15 cells. However, CSE significantly increased the viability of alloxan-treated cells and lowered LDH leakage. The intracellular NAD+/NADH ratio and insulin secretion were also significantly increased by 1.7-fold and 1.3-fold, respectively, after treatment with 100 ㎍/㎖ CSE. The HIT-T15 cells treated with alloxan showed significant decreases in the activities of antioxidant enzymes, while CSE significantly elevated the levels of antioxidant enzymes. These findings suggest that CSE could have a protective effect against cytotoxicity and dysfunction of pancreatic cells in the presence of alloxan-induced oxidative stress.
The effect of water extract in fruits of Omija (Schizandra chinensis Baillon) on alloxan-induced diabetic rats were determining the contents of metabolites and enzyme activities in the liver and serum of rat. The treatment with water extract in fruits of Omija showed increased in contents of protein, glycogen, and activity of glucose-6-phosphate dehydrogenase and a decreased in pyruvate content in the hepatic tissue from the alloxan treated group. These treatments were decreased contents of glucose, urea nitrogen, free fatty acid and activities of GPT, GOT, LDH.
The possible effects of 10% carbohydrate containting diets on the serum glucose and lipids values were studied with normal and alloxan - diabetic male rats for a period of 28 days. The diets were supplemented with either glucose, cornstarch or sucrose, fructose or the basal diet containing no added carbohy drates. After this period, 3 to 4 rats among the each group were killed and samples of serum, liver, spleen and kidney were collected. The others were rendered diabetic by the intraperitoneal injection of alloxan (190mg/wtkg). Then original feeding scedule was continued for 3 days in all five diabetic groups, before the collection of tissue samples and serum. Feeding the nondiabetic rats with glucose and fructose delayed the bodyweight development relatively compared with nonsugar group. The weight deductions after alloxan injection were similar in all feeding groups. All diabetic animals exhibited increased blood glucose triglceride levels but almost unchanged total cholesterol values. Blood glucose values for nondiabetic rats were normal ranges, and then glucose feeding group was the highest. Total - cholesterol values were the highest in nondiabetic rats fed glucose, fructose and in diabetics fed sucrose, glucose. Triglyceride values were the highest in nondiabetic rats fed sucrose, and no difference in diabetic rats. Some cornstarch group exhibited fatty-livers in diabetic and nondiabetic, and more studies need.
본 연구는 국내에서 상용 중인 과일 및 채소로부터 췌장베타세포 보호활성을 가지는 추출물의 식물 자원을 탐색해보고자 수행되었다. Alloxan에 대한 총 13종의 과일・채소 추출물에 대한 효과를 햄스터췌장베타세포주(HIT-T15)를 배양한 후 세포생존율, LDH 방출량, NAD+/NADH ratio, 인슐린 분비량 및 4종의 항산화효소활성을 측정하였다. 총 13종의 과일・채소 추출물은 alloxan에 의해 감소된 세포생존율을 유의하게 증가시켰다. 마늘종(Allium sativum) 등의 11종의 시료는 alloxan에 의해 증가된 LDH 방출량을 유의적으로 감소시켰으며, 산수유(Cornus officinalis) 등의 9종의 시료는 alloxan에 의해 감소된 NAD+/NADH ratio를 유의적으로 상승시킴으로써 세포생존율을 증가시켰다. 또한 3종의 추출물인 마늘종(Allium sativum), 산수유(Cornus officinalis)와 고들빼기(Yongjia sonchifolia) 추출물은 인슐린 분비능을 보호하였고 깻잎 등 5종의 시료는 SOD, GST, GR 및 GPx와 같은 항산화효소 활성을 유의적으로 증가시켰다. 과일・채소 추출물은 세포괴사 및 DNA fragmentation을 억제하고 세포 내 항산화효소 활성을 증가시킴으로써 alloxan에 의해 유발된 산화스트레스로부터 췌장베타세포를 보호하는 것으로 생각된다. 이상의 연구결과로부터 마늘종(Allium sativum), 산수유(Cornus officinalis), 자두(Prunus salicina), 오미자(Schisandra chinensis) 및 고들빼기(Yongjia sonchifolia)와 같은 5종의 추출물이 alloxan에 의한 산화스트레스로부터 췌장베타세포를 보호하는 효과가 우수한 것으로 판단되며 향후 췌장베타세포의 손상을 억제하는 기능성 식품 등의 개발에 기초 연구자료로 활용될 것으로 생각된다.
본 연구는 췌장베타세포인 HIT-T15 세포를 이용하여 매실주정추출물(PME)의 alloxan에 의한 산화스트레스로부터의 세포보호, 인슐린 분비능 및 항산화 효소 활성을 평가하였다. PME는 alloxan에 의해 유발된 산화스트레스로부터 세포를 보호하여 세포생존율을 증가시켰다. PME는 세포막 손상지표인 LDH 방출을 억제하였고 NAD+/NADH ratio를 유의적으로 증가시켜 세포사멸이 억제되어짐을 확인하였다. 또한 alloxan 단독처리군에 비해 250 μg/ml PME 처리군에서 인슐린 분비량이 유의성 있게 증가하였다. Alloxan과 PME를 동시에 처리하여 HIT-T15세포의 항산화효소 활성을 측정했을 때 산화스트레스에 의해 감소되었던 항산화효소 활성이 PME에 의해 보호되는 효과를 확인하였다. 이상의 연구결과로부터 PME는 세포괴사 및 DNA fragmentation을 억제하고 세포내 항산화효소 활성을 증가시켜 alloxan에 의해 유발된 산화스트레스로부터 췌장베타세포를 보호하고 이에 따른 인슐린 분비능 조절 효과가 있는 것으로 생각된다.