摘要
目的:基于血管紧张素转换酶1(ACE1)/血管紧张素Ⅱ(AngⅡ)/血管紧张素Ⅱ1型受体(AT1R)轴,探讨六味地黄汤加减防治糖尿病肾病(DKD)肾脏损伤的可能作用机制。方法:随机将50只雄性SD大鼠分为正常组8只,造模组42只,造模组大鼠高糖高脂饲料喂养6周后,予一次性腹腔注射链脲佐菌素(STZ)35 mg·kg^(-1)诱导建立DKD大鼠模型。造模成功后随机分为模型组、中药组(六味地黄汤加减颗粒剂21 g·kg^(-1))、西药组(氯沙坦钾33 mg·kg^(-1))、中西药组(氯沙坦钾33 mg·kg^(-1)联合六味地黄汤加减颗粒剂21 g·kg^(-1)),连续灌胃8周后检测各组大鼠空腹血糖(FBG)、尿蛋白(Up)、尿素氮(Bun)、血肌酐(SCr);酶联免疫吸附测定法(ELISA)检测血清ACE1、AngⅡ、AT1R水平;蛋白免疫印迹法(Western blot)检测各组大鼠肾组织ACE1、AngⅡ、AT1R蛋白表达水平;苏木素-伊红(HE)、马松(Masson)、过碘酸雪夫氏(PAS)染色后观察肾组织病理形态学改变;采集各组大鼠粪便标本进行16S rDNA高通量测序分析。结果:与正常组比较,模型组Up、FBG、Bun、SCr、ACE1、AngⅡ、AT1R水平显著升高(P<0.01),肾组织病变严重,ACE1、AngⅡ、AT1R蛋白表达显著增加(P<0.01),厚壁菌门/拟杆菌门(F/B)升高,乳杆菌属的物种丰度下降,穆氏菌属、双歧杆菌属的物种丰度升高;与模型组比较,各给药组Bun、SCr、ACE1、AngⅡ、AT1R水平均显著下降(P<0.01),肾脏病变程度有所改善,中药组、中西药组Up、FBG水平均显著下降(P<0.01),西药组和中西药组的ACE1、AngⅡ、AT1R蛋白,中药组的AngⅡ蛋白表达显著减少(P<0.01)、中药组ACE1、AT1R蛋白表达明显减少(P<0.05),中药组、中西药组的F/B降低;西药组和中药组布劳特氏菌属的物种丰度升高,中药组、中西药组乳杆菌属、瘤胃球菌科未定属、双歧杆菌属的物种丰度升高,穆氏菌属的物种丰度下降,中药组、中西药组Chao 1和Ace指数明显增加(P<0.05);与西药组比
Objective:To explore the mechanism of modified Liuwei Dihuangtang in preventing and treating renal injury in diabetic kidney disease(DKD)via the angiotensin-converting enzyme 1(ACE1)/angiotensinⅡ(AngⅡ)/angiotensin Ⅱ type 1 receptor(AT1R)axis.Method:Fifty male SD rats were randomized into a normal group(n=8)and a modeling group(n=42).The rats in the modeling group were fed with a high-sugar and high-fat diet for 6 weeks and intraperitoneally injected with 35 mg·kg^(-1) streptozotocin(STZ)to establish the model of DKD.After successful modeling,the rats were randomized into model,traditional Chinese medicine(modified Liuwei Dihuangtang granules 21 g·kg^(-1)),western medicine(losartan potassium,33 mg·kg^(-1)),and integrated Chinese and western medicine(losartan potassium 33 mg·kg^(-1) combined with modified Liuwei Dihuangtang granules 21 g·kg^(-1))groups.The levels of fasting blood glucose(FBG),urinary protein(Up),blood urea nitrogen(Bun),and serum creatinine(SCr)were measured in each group after 8 consecutive weeks of drug intervention.Enzyme-linked immunosorbent assay was employed to determine the serum levels of ACE1,AngⅡ,and AT1R.Western blot was employed to measure the protein levels of ACE1,AngⅡ,and AT1R in the renal tissue.The pathological and morphological changes of the renal tissue were observed after hematoxylin-eosin(HE)staining,Masson staining,and periodic acid Schiff's(PAS)staining.The fecal samples of rats in each group were collected for 16S rDNA high-throughput sequencing.Result:Compared with the normal group,the model group showed elevated levels of Up,FBG,Bun,SCr,ACE1,AngⅡ,and AT1R(P<0.01),serious lesions in the renal tissue,up-regulated protein levels of ACE1,AngⅡ,and AT1R(P<0.01),increased Firmicutes/Bacteroidetes(F/B)ratio,decreased relative abundance of Lactobacillus,and increased relative abundance of Moralella and Bifidobacteria.Compared with the model group,drug intervention lowered the levels of Bun,SCr,ACE1,AngⅡ,and AT1R(P<0.01)and alleviated the pathological changes i
作者
杨超茅
张顺宵
李园园
高建东
YANG Chaomao;ZHANG Shunxiao;LI Yuanyuan;GAO Jiandong(Baoshan Branch,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine(TCM),Shanghai 201999,China;Baoshan Hospital Affiliated to Shanghai University of TCM,Shanghai 201999,China;Shuguang Hospital Affiliated to Shanghai University of TCM,Shanghai 201203,China;Key Laboratory of Liver and Kidney Diseases,Ministry of Education,Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Nephropathy Institute of Chinese Medicine,Shanghai University of TCM,Shanghai 201203,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第6期1-9,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金青年项目(82004117)
上海市卫生健康委员会科研课题(20214Y0195)
宝山区科学技术委员会科技创新专项资金医学卫生项目(20-E-14)
宝山区卫生健康委优青(育才)计划项目(BSWSYC-2023-05)。
关键词
血管紧张素转换酶1
血管紧张素Ⅱ
血管紧张素Ⅱ1型受体
肠道菌群
六味地黄汤加减
angiotensin-converting enzyme 1(ACE1)
angiotensinⅡ(AngⅡ)
angiotensinⅡtype 1 receptor(AT1R)
intestinal flora
modified Liuwei Dihuangtang
