天然产物研究与开发 ›› 2023, Vol. 35 ›› Issue (12): 2040-2048.doi: 10.16333/j.1001-6880.2023.12.003

• 研究论文 • 上一篇    下一篇

基于肠道菌群探讨气阴两虚证糖尿病肾脏疾病动物模型

陈鹏德1,姚   蓝1*,郭   凤1,韩   雪1,张文祥1,韩   荣2*   

  1. 1新疆医科大学中医学院;2新疆医科大学教务科,乌鲁木齐 830054
  • 出版日期:2023-12-28 发布日期:2023-12-27
  • 基金资助:
    新疆维吾尔自治区自然科学基金(2018D01C176)

Study on the animal model of diabetic kidney disease with Qi-yin deficiency syndrome based on intestinal flora

CHEN Peng-de1,YAO Lan1*,GUO Feng1,HAN Xue1,ZHANG Wen-xiang1,HAN Rong2*   

  1. 1College of Traditional Chinese Medicine,Xinjiang Medical University; 2Department of Educational Affairs,Xinjiang Medical University,Urumqi 830054,China
  • Online:2023-12-28 Published:2023-12-27

摘要:

建立气阴两虚病证糖尿病肾脏疾病病证结合动物模型及评价指标。将30只Wistar大鼠按照体重随机分为正常组(10只)、模型组(20只)。模型组采用高糖高脂饲料喂养4周,腹腔注射链脲佐菌素(streptozotocin,STZ)25 mg/kg诱导形成糖尿病大鼠。把成模大鼠按照24 h尿蛋白值分配到糖尿病肾脏疾病模型组(10只)、气阴两虚病证结合模型组(10只),气阴两虚病证结合模型组以附子、青皮、枳实按照5∶6∶6,13 g/kg灌胃诱导8周。对大鼠各项测量指标进行观测。发现与正常组和糖尿病肾脏疾病模型组比较,气阴两虚病证结合模型组大鼠粪便中有益菌:乳酸杆菌属、梭菌属、拟杆菌门、双歧杆菌属含量显著降低;致病菌:梭杆菌属、肠杆菌、毛螺菌科、粪肠球菌含量显著升高;气阴两虚病证结合模型组大鼠血清中免疫球蛋白G(IgG)(0.5±0.1 g/L)、免疫球蛋白M(IgM)(14.2±0.6 g/L)和肾脏组织中钠钾泵酶(Na+/K+-ATP)(0.8±0.3 U/mg)、钙镁泵酶(Ca2+-Mg2+-ATP)(2.0±0.3 U/mg)显著低于正常组(P<0.05),环磷酸腺苷(cAMP)(0.9±0.2 mmol/L)、环磷酸腺苷/环磷酸鸟苷(cGMP)(0.3±0.1 mmol/L)显著高于正常组(P<0.05)。HE染色显示:与正常组和糖尿病肾脏疾病模型组比较,气阴两虚病证结合模型组大鼠出现明显肾脏和结肠损伤;肾脏和结肠组织的肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)蛋白表达均显著升高,结肠中毒蕈碱型胆碱受体M3(M3R)、5-羟色胺受体3A(5-HT3A)、闭合蛋白(Occludin)、紧密连接蛋白(ZO-1)表达显著降低。实验结果表明通过高糖高脂喂养4周联合腹腔注射STZ 25 mg/kg、附子、青皮、枳实灌胃诱导大鼠8周,可部分形成气阴两虚证糖尿病肾脏疾病基本证候特征,用于评价气阴两虚证糖尿病肾脏疾病的建立。

关键词: 糖尿病肾病, 气阴两虚, 病证结合, 肠道菌群

Abstract:

To establish an animal model and evaluation index of Qi-yin deficiency syndrome of diabetic kidney disease.Thirty Wistar rats were randomly divided into normal group (10 rats) and model group (20 rats) according to body weight.The model group was fed high-carbohydrate and high-fat diet for four weeks,and the diabetic rats were induced by intraperitoneal injection of streptozotocin (STZ) 25 mg/kg.The model rats were assigned to diabetic kidney disease model group (10 rats) and Qi-yin deficiency syndrome combined model group (QYCM) (10 rats) according to the 24 h urinary protein value.The QYCM was administrated with Aconm Lateralis Radix Praeparaia,Pericarpium Citri Peticulatae Viride and Fructus Aurantii Immaturus by gavage at 5∶6∶6,13 g/kg for eight weeks.The measurement indexes of rats were observed.It was found that compared with normal group and diabetic kidney disease model group,the content of beneficial bacteria in fecal stool of rats in combination with QYCM:Lactobacillus, Clostridium,Bacteroides and Bifidobacterium decreased significantly;Pathogenic bacteria:Fusobacterium,Enterobacter,Trichospirillaceae,Enterococcus faecalis increased significantly;Serum immunoglobulin G(IgG),immunoglobulin M(IgM) and kidney tissue sodium potassium pump enzyme (Na+/K+-ATP),calcium magnesium pump(Ca2+-Mg2+-ATP) in kidney tissue were significantly lower than those in normal group (P<0.05).Cyclic adenosine phosphate (cAMP) and cyclic adenosine phosphate/cyclic guanosine phosphate (cGMP) were significantly higher than those in normal group (P<0.05).HE staining showed that compared with normal group and diabetic kidney disease model group,there were obvious kidney and colon injuries in QYCM.The expressions of tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) in kidney and colon were significantly increased,while the expressions of muscarinic choline receptor M3 (M3R),5-hydroxytryptamine receptor 3A(5-HT3A),occludin and tight junction protein (ZO-1) in colon were significantly decreased.The experimental results showed that the basic syndrome characteristics of diabetic kidney disease in Qi-yin deficiency syndrome could be partially formed by feeding rats with high sugar and high fat for four weeks combined with abdominal injection of STZ 25 mg/kg,Aconm Lateralis Radix Praeparaia,Pericarpium Citri Peticulatae Viride and Fructus Aurantii Immaturus for eight weeks,which was used to evaluate the establishment of diabetic kidney disease in Qi-yin deficiency syndrome.

Key words: diabetic kidney disease, Qi-yin deficiency, combination of disease and syndrome, intestinal flora

中图分类号:  R285