CAJ | 학술논문

目的探讨高压氧对糖尿病大鼠胰岛细胞凋亡和超微结构的影响.方法 8只Wistar大鼠为正常对照组(对照组),空腹血糖16.7 mmol/L以上的24只Goto-Kakizaki (GK)大鼠,根据血糖高低排序后采用数字表法分为模型组、二甲双胍组、高压氧组,每组8只.对照组、模型组、高压氧组每天按5 ml/kg灌服纯净水,二甲双胍组按250 mg/kg灌服二甲双胍混悬液,高压氧组每天1次高压氧治疗.3周后,各组大鼠用异氟醚麻醉后取胰腺,部分组织制成电镜切片观察,部分组织制成石蜡切片后分别进行苏木素-伊红(HE)染色、原位末端标记法(TdT-mediated dUTP nick-end labeling,TUNEL)法检测胰岛细胞凋亡,免疫组织化学二步法对胰岛进行半胱氨酸天冬氨酸蛋白酶-3(caspase-3)标记.结果电镜观.察到模型组胰岛细胞线粒体明显肿胀、脱颗粒、呈空泡状,高压氧组胰岛细胞线粒体病变较模型组病变轻.各组均有胰岛细胞凋亡发生,高压氧组凋亡指数(AI)为(1.57±0.33)％,caspase-3阳性细胞积分光密度值(IOD)为10.22 ±4.61,模型组AI和IOD值分别为(4.88±0.51)％和14.65 ±4.8,2组比较差异均有统计学意义(P＜0.05),与对照组[AI为(1.16±0.31)％,IOD值为5.36 ±2.34]、二甲双胍组[AI为(1.72±0.14)％,IOD值为9.37 ±4.64]比较,差异均无统计学意义(P＞0.05).结论高压氧可保护线粒体膜,减轻高血糖对线粒体的损伤,抑制2型糖尿病大鼠胰岛细胞的凋亡和caspase-3抗原的表达,有利于糖尿病的治疗.
목적 탐토고압양대당뇨병대서이도세포조망화초미결구적영향.방법 8지Wistar대서위정상대조조(대조조),공복혈당16.7 mmol/L이상적24지Goto-Kakizaki (GK)대서,근거혈당고저배서후채용수자표법분위모형조、이갑쌍고조、고압양조,매조8지.대조조、모형조、고압양조매천안5 ml/kg관복순정수,이갑쌍고조안250 mg/kg관복이갑쌍고혼현액,고압양조매천1차고압양치료.3주후,각조대서용이불미마취후취이선,부분조직제성전경절편관찰,부분조직제성석사절편후분별진행소목소-이홍(HE)염색、원위말단표기법(TdT-mediated dUTP nick-end labeling,TUNEL)법검측이도세포조망,면역조직화학이보법대이도진행반광안산천동안산단백매-3(caspase-3)표기.결과 전경관.찰도모형조이도세포선립체명현종창、탈과립、정공포상,고압양조이도세포선립체병변교모형조병변경.각조균유이도세포조망발생,고압양조조망지수(AI)위(1.57±0.33)％,caspase-3양성세포적분광밀도치(IOD)위10.22 ±4.61,모형조AI화IOD치분별위(4.88±0.51)％화14.65 ±4.8,2조비교차이균유통계학의의(P＜0.05),여대조조[AI위(1.16±0.31)％,IOD치위5.36 ±2.34]、이갑쌍고조[AI위(1.72±0.14)％,IOD치위9.37 ±4.64]비교,차이균무통계학의의(P＞0.05).결론 고압양가보호선립체막,감경고혈당대선립체적손상,억제2형당뇨병대서이도세포적조망화caspase-3항원적표체,유리우당뇨병적치료.
Objective To investigate the effects of hyperbaric oxygen (HBO) on the apoptosis and ultrastructure of islet cells in rats with type 2 diabetes.Methods Twenty-four Goto-Kakizaki (GK) rats with fasting blood glucose of over 16.7 mmol/L were randomly assigned to the 3 groups:the model group,the metformin hydrochloride group,and the HBO group,each consisting of 8 animals.Another 8 healthy male Wistar rats were used as the control group.The control group,the model group and the HBO group were given 5 ml/kg of pure water a day.The metformin hydrochloride group had lavage of 250 mg/kg metformin hydrochloride solution a day,and the HBO group had one session of HBO treatment a day.After 3 weeks,following anesthesia with isoflurane,the pancreas tissue was taken from the rats of all the groups,some of the tissue was made into slides for electron-microscopic observation,and some of them was made into paraffin sections for staining with Hematoxylin-eosin (HE).Apoptosis of islet cells was detected with immunohistochemistry with TdT-mediated dUTP Nick-End Labeling (TUNEL),and islet cells were labeled with caspase-3 by using immunohistochemistry.Results Electron microscopy revealed that mitochondria in the animals of the model group was markedly swollen with degranulation,and vacuolation.Pathological changes inmitochondria of islet cells for the animals of the HBO group were lighter than those of the model group.Apoptosis of islet cells occurred in all the groups,with the apoptosis index (AI) of the HBO group being (1.57 ± 0.33) ％.IOD for caspase-3 positive cells was 10.22 ± 4.61.On the other hand,AI and IOD for the animals of the model group were (4.88 ± 0.51) ％ and 14.65 ± 4.8 respectively,and statistical significance could be seen,when comparisons were made between the 2 groups(P ＜ 0.05).No statistical significance could be noted,as compared with the data of AI (1.16 ± 0.31)％ and the data of IOD (5.36 ± 2.34) in the control group,and the data of AI(1.72 ± 0.14)％ and the data of IOD (9.37 ± 4.64) in the metformin hydrochloride group (P ＞ 0.05).Conclusions HBO could protect the mitochondrial membrane,alleviate mitochondrial lesion induced by hyperglycemia,and furthermore could inhibit apoptosis of islet cells in the rats with type 2 diabetes,and the expression of caspase-3 as well.Therefore,it would be beneficial for treatment of diabetes mellitus.