青钱柳叶水提物对2型糖尿病大鼠胰腺β细胞凋亡的影响  

Effects of Cyclocarya Paliurus leaves aqueous extract on pancreatic β cell's apoptosis in type 2 diabetic rats

作　　者：姚骏凯 王景霞[1] 高学敏[1] 付璐 高飞[1] 王玉杰[1] 贾岚 王蕾蕾 Yao Junkai;Wang Jingxia;Gao Xuemin;Fu Lu;Gao Fei;Wang Yujie;Jia Lan;Wang Leilei(Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学,北京100029

出　　处：《北京中医药大学学报》2018年第8期663-669,共7页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(No.81673617)~~

摘　　要：目的探讨青钱柳叶水提物对2型糖尿病(T2D)大鼠胰腺β细胞凋亡的影响。方法采用高脂饲料喂养8周,并单次腹腔注射链脲佐菌素建立T2D大鼠模型。造模成功后,将模型大鼠随机分为模型组,二甲双胍组,青钱柳叶水提物低、中、高剂量组。连续灌胃给药28 d,正常组、模型组(生理盐水),二甲双胍组(0.30 g/kg),青钱柳叶水提物低、中、高剂量组(0.18、0.36、1.08 g/kg),末次给药时检测大鼠糖耐量。处死大鼠,摘取胰腺,比色法测超氧化物歧化酶(SOD)、过氧化酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)、丙二醛(MDA)、游离脂肪酸(FFA)含量;放免法测肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)含量;免疫酶联吸附法(ELISA)测可溶性细胞凋亡因子(Fas)、可溶性细胞凋亡因子配体(Fasl)、活化半胱氨酸蛋白酶8(Caspase8)含量;Western bolt法检测总c-Jun氨基末端激酶1(T-JNK1)、磷酸化c-Jun氨基末端激酶1(P-JNK1)、胰腺十二指肠同源盒子1(PDX-1)蛋白表达水平;TUNEL染色观察胰腺细胞凋亡情况。结果青钱柳叶水提物可提高模型大鼠胰腺SOD、CAT、GSH-PX含量及PDX-1蛋白表达量;降低模型大鼠空腹血糖、曲线下面积,胰腺MDA、FFA,TNF-α、IL-6、Fas、Fasl、Caspase8的含量,T-JNK1、p-JNK1的表达量,胰腺细胞凋亡率。结论青钱柳叶水提物能提高胰腺抗氧化性,缓解氧化应激,抑制胰腺β细胞凋亡;其机制可能与抑制T-JNK1、P-JNK1蛋白的表达,提高PDX-1蛋白的表达,降低Fas、Fasl、Caspase8含量有关。Objective To investigate the effects of Cyclocarya Paliurus aqueous extract（ CPAE） on pancreatic β cell apoptosis in type 2 diabetic（ T2 D） rats. Methods T2 D ract model was established by high-fat diet feeding for 8 weeks and single intraperitoneal injection of streptozotocin. From the ninth week,the rats in the model group（ sodium chloride solution）,metformin group（ 0. 3 g/kg）,low,medium and high dose CPAE groups（ 0. 18,0. 36,1. 08 g/kg） were treated for 4 consecutive weeks. Glucose tolerance was measured at the last administration. After sacrifing the rats and removing the pancreas,the contents of superoxide dismutase（ SOD）,peroxidase（ CAT）,glutathione peroxidase（ GSHPX）,malondialdehyde（ MDA） and free fatty acid（ FFA） were detected with colorimetric method.Tumor necrosis factor alpha（ TNF-α） and interleukin 6（ IL-6） were measured with radioimmunoassay.The levels of apoptosis factor（ Fas） and its ligand（ Fasl）,activated cysteine protease（ Caspase8） were detected by using ELISA. Western blot assay was used to detect total c-Jun N-terminal kinase 1（ TJNK1）,phosphorylated c-Jun N-terminal kinase 1（ P-JNK1）,pancreatic duodenal homeobox 1（ PDX-1） protein expression; TUNEL staining was used to observe pancreatic cell＇s apoptosis. Results CPAE treated groups presented with increased contents of SOD,CAT,GSH-PX and increased expression of PDX-1 in the pancreas of the CPE groups; fasting blood glucose,area under the curve,MDA,FFA,TNFα,IL6,Fas,Fasl,Caspase8 content,T-JNK1,p-JNK1 expression,and pancreatic cell apoptosis rate were all reduced in the model group. Conclusion CPAE can improve the pancreas antioxidant,alleviate oxidative stress,and inhibit pancreatic β cell＇s apoptosis. The mechanism may be related to inhibiting the expression of T-JNK1 and P-JNK1 protein,increasing the expression of PDX-1 protein and decreasing the content of Fas,Fasl and Caspase8.

关 键 词：青钱柳叶 氧化应激 c-Jun氨基末端激酶1 胰腺十二指肠同源盒子1 Β细胞凋亡 大鼠 

分 类 号：R285.5[医药卫生—中药学]