首页|基于自噬调控探讨解毒通络调肝方对SD大鼠胰岛细胞保护的作用机制

基于自噬调控探讨解毒通络调肝方对SD大鼠胰岛细胞保护的作用机制

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观察解毒通络调肝方基于自噬调控保护2型糖尿病SD大鼠胰岛细胞的作用机制.8周龄SD雄性大鼠,高脂饲料喂养4周后联合链脲佐菌素40 mg/kg单次腹腔注射诱导2型糖尿病大鼠动物模型,中药组予解毒通络调肝方低剂量(1.5 g·kg-1·d-1)、中剂量(4.5 g·kg-1·d-1)及高剂量(13.5 g·kg-1·d-1),西药组予盐酸二甲双胍(0.15 g·kg-1·d-1).连续灌胃4周,检测各组大鼠空腹血糖、糖化血清蛋白、甘油三酯、胆固醇、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇、游离脂肪,ELISA法检测空腹血清胰岛素并计算胰岛素抵抗指数,进行葡萄糖耐量试验.取SD大鼠胰腺组织,HE染色,免疫组织化学法检测Beclin-1、LC3蛋白的表达,Western blot法检测Beclin-1、LC3、P62蛋白的表达,RT-PCR法检测Beclin-1、LC3RNA的表达.结果显示,解毒通络调肝方及盐酸二甲双胍组均有较好的降糖及降脂效果(P<0.05,P<0.01).胰腺组织HE染色显示,结构紊乱,边界不清,排列不规则,胞浆少,胰岛内细胞质染色变浅或呈空泡化现象,在解毒通络调肝方及盐酸二甲双胍组中明显改善.自噬调控方面,运用免疫组织化学法、Western blot法、RT-PCR法检测,解毒通络调肝方及盐酸二甲双胍组对自噬标志蛋白Beclin-1及LC3表达均较模型组显著上升(P<0.01);降低P62蛋白表达(P<0.05,P<0.01).结果证明,解毒通络调肝方可有效缓解2型糖尿病SD大鼠体质量下降趋势,改善多饮情况,具有较好地改善糖、脂代谢的作用,并且可缓解2型糖尿病SD大鼠胰腺组织病理结构的损害保护胰岛细胞,其作用可能是通过调控自噬水平实现的.
Exploring the protective mechanism of Jiedu Tongluo Tiaogan decoction on pan-creatic islet cells in SD rats based on autophagy regulation
The aim of this study is to explore the protective mechanism of Jiedu Tongluo Tiaogan decoction(JTTD)on islet cells of type 2 diabetic SD rats based on autophagy regulation.8-week-old SD male mice were fed with high-fat diet for 4 weeks and then induced with a single intraper-itoneal injection of streptozotocin 40 mg/kg.The Chinese medicine group was given the low(1.5 g·kg-1·d-1),medium(4.5 g·kg-1·d-1)and high(13.5 g·kg-1·d-1)doses of JTTD,and the western medicine group was given metformin hydrochloride(0.15 g·kg-1·d-1).Fasting blood glucose(FBG),glycated serum protein(GSP),triglycerides(TG),cholesterol(CHO),high-densi-ty lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),free fat(FFA)were measured.Moreover,the fasting serum insulin was determined by ELISA method and insulin resistance index(HOMA-IR)was calculated,glucose tolerance test was performed.The pancreatic tissues of SD mice were stained with HE,and the expression of Beclin-1 and LC3 were checked by immunohistochemistry,the expression of Beclin-1,LC3 and P62 was determined by Western blot,the expression of Beclin-1 and LC3RNA was determined by RT-PCR.The results showed that both the JTTD group and metformin hydrochloride group had better hypoglycemic and lipid-lowering effects(P<0.05,P<0.01).HE staining of pancreatic tissues showed that disorganized structure,unclear boundary,irregular arrangement,less cytoplasm,and lighter cytoplasmic staining or vacuo-lation in the islets in the JTTD group and metformin hydrochloride group were improved.In terms of autophagy regulation,immunohistochemistry,Western blot and RT-PCR,compared with the model group,the expression of Beclin-1 and LC3 mRNA was significantly increased(P<0.01)and the level of p62 protein was decreased(P<0.05,P<0.01)in the JTTD group and metformin hydrochloride group.In conclusion,the JTTD can alleviate the trend of weight loss and improve ex-cessive drinking effectively in type 2 diabetic mellitus SD mice,as well as have better effects on im-proving glucose and lipid metabolism,moreover,it can alleviate the damage of pancreatic his-topathological structure and protect islet cells,which may be achieved by regulating the level of au-tophagy.

type 2 diabetes mellitusJiedu Tongluo Tiaogan decoctionautophagyislet cells

金美英、潘韦韦、苏婧、王婉霖、崔镇海

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长春中医药大学附属第三临床医院,吉林长春 130117

长春医学高等专科学校临床医学院,吉林长春 130033

长春中医药大学,吉林长春 130021

2型糖尿病 解毒通络调肝方 自噬 胰岛细胞

2024

中国兽医学报
吉林大学

中国兽医学报

CSTPCD北大核心
影响因子:0.702
ISSN:1005-4545
年,卷(期):2024.44(11)