摘要
目的:探讨补阳还五汤是否通过调控核转录因子E2相关因子2(Nrf2)/血红素加氧酶-1(HO-1)通路抑制糖尿病肾病(DKD)小鼠氧化应激,从而发挥减轻肾组织损伤,延缓肾脏纤维化的作用。方法:将73只C57BL/6小鼠随机分为造模组63只和正常对照组10只,造模组小鼠高糖高脂饲料喂养6 w后,连续5 d以50 mg/kg腹腔注射链脲佐菌素(STZ)制备糖尿病模型后继续高糖高脂饲料喂养8 w,当小鼠出现尿蛋白阳性则DKD模型制备成功。随机将DKD小鼠分为模型对照组、罗格列酮7.05×10^(-4)g/kg组、补阳还五汤3.21、6.41、12.82 g/kg组,各组灌胃相应药物或生理盐水,1次/d,连续8 w。药物干预期间监测小鼠体质量、血糖、尿量、24 h尿蛋白定量(24 h-UTP),给药结束后采用马松(Masson)染色及高碘酸希夫(PAS)染色观察小鼠肾组织病理学改变;测定超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活力;荧光探针标记法检测小鼠肾组织活性氧簇(ROS)水平;蛋白免疫印迹法(Western blot)检测小鼠肾组织转化生长因子β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)、纤连蛋白(FN)、Ⅰ型胶原(CollagenⅠ)、Nrf2、HO-1蛋白表达。结果:与正常对照组比较,模型对照组小鼠体质量下降,血糖升高,尿量、24 h-UTP增加,肾小球囊壁增厚,有明显的胶原纤维增生,肾小球内系膜增生,肾小球周围可见糖原颗粒沉积,SOD、CAT活力降低,ROS水平增加,Nrf2、HO-1蛋白表达下调,TGF-β1、α-SMA、FN、CollagenⅠ蛋白表达上调(P<0.05或P<0.01);与模型对照组比较,补阳还五汤各组及罗格列酮7.05×10^(-4)g/kg组小鼠体质量增加,尿量、24 h-UTP降低,肾组织病理学损伤可见不同程度改善,SOD、CAT活力明显增加,ROS水平明显降低,Nrf2、HO-1表达明显上调,TGF-β1、α-SMA、FN、CollagenⅠ表达明显下调(P<0.05或P<0.01)。结论:补阳还五汤可通过激活Nrf2/HO-1通路抑制氧化应激,缓解DKD小鼠肾纤维化,减轻小鼠病理损伤。
Objective:To explore whether Buyang Huanwutang(补阳还五汤)inhibits oxidative stress in diabetic kidney disease(DKD)mice by regulating nuclear transcription factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)pathway,to alleviate kidney tissue damage and delay kidney fibrosis.Methods:A total of 73 C57BL/6 mice were randomly divided into model group(n=63)and normal control group(n=10).Mice in model group were fed with high-sugar and high-fat diet for 6 weeks,and after intraperitoneal injection of 50 mg/kg streptozotocin(STZ)for 5 days,and then,they were continuously given high-sugar and high-fat diet for 8 weeks.When the mice showed positive urine protein,DKD model was successfully prepared.The DKD mice were randomly divided into model control group,rosiglitazone 7.05×10^(-4)g/kg group,and Buyang Huanwutang 3.21,6.41 and 12.82 g/kg groups.Mice in all groups were given corresponding drugs or normal saline,once per day for 8 consecutive weeks.During the drug intervention period,the body mass,blood glucose,urine volume and 24-hour urinary total protein quantity(24 h-UTP)of the mice were monitored.After administration,Masson staining and Periodic Acid-Schiff(PAS)staining were used to observe the pathological changes in mouse kidney tissue.The activities of superoxide dismutase(SOD)and catalase(CAT)were measured.The expression of reactive oxygen species(ROS)in mouse kidney tissue was detected by fluorescence probe labeling,and the protein expressions of transforming growth factorβ1(TGF-β1),α-smooth muscle actin(α-SMA),fibronectin(FN),collagenⅠ(ColⅠ),Nrf2 and HO-1 were detected by Western blot.Results:Compared with the normal control group,the model group had decreased body mass,increased blood sugar,elevated urine volume and 24 h-UTP,thickened glomerular capsule wall,significant collagen fibrous dysplasia,mesangial hyperplasia in the glomerulus,deposition of glycogen particles around the glomerulus,reduced activities of SOD and CAT,enhanced expression of ROS,down-regulated expressions of Nrf2 and HO-1,and up-regulated expressions of TGF-β1,α-SMA,FN and CollagenⅠ(P<0.05 or P<0.01).Compared with the model control group,mice in Buyang Huanwutang groups and rosiglitazone group gained weight,and their blood sugar,urine volume and 24 h-UTP were decreased.The pathological damage of kidney tissue was improved to varying degrees,and the activities of SOD and CAT were increased.Additionally,the expressions of Nrf2 and HO-1 was up-regulated,while the expressions of ROS as well as TGF-β1,α-SMA,FN,and CollagenⅠwere down-regulated(P<0.05 or P<0.01).Conclusion:Buyang Huanwutang can inhibit oxidative stress by activating Nrf2/HO-1 pathway,thus alleviating kidney fibrosis in DKD mice and relieving the pathological damage.
作者
郑琳琳
边东
郭登洲
ZHENG Linlin;BIAN Dong;GUO Dengzhou(Graduate School,Hebei University of Chinese Medicine,Shijiazhuang 050091;The First Affiliated Hospital of Hebei University of Chinese Medicine,Shijiazhuang 050011)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2024年第4期42-47,共6页
Pharmacology and Clinics of Chinese Materia Medica
基金
河北省自然科学基金项目(编号:H2022423367)
河北中医学院研究生创新能力培养资助项目(编号:XCXZZBS2023015)。
关键词
补阳还五汤
糖尿病肾病
核转录因子E2相关因子2/血红素加氧酶-1通路
氧化应激
纤维化
Buyang Huanwutang(补阳还五汤)
Diabetic kidney disease
Nuclear transcription factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)pathway
Oxidative stress
Fibrosis
