目的 研究表没食子儿茶素没食子酸酯((-)-Epigallocatechin gallate,EGCG)对链脲佐菌素(Streptozocin,STZ)诱导的糖尿病大鼠肾脏的保护作用及其作用机制。方法 30只SD雄性大鼠随机分为正常对照组、模型组、EGCG剂量组(10、20、40 mg·kg-1·d-1),后4组大鼠一次性腹腔注射STZ(65 mg·kg-1)建立糖尿病大鼠模型。于造模成功后第一天开始,EGCG各给药组大鼠按上述剂量灌胃给药,正常对照组和模型组大鼠均给予三蒸水10 mL·kg-1·d-1。药物治疗后第6周和第12周测量各组大鼠的体重、饮水量和摄食量。12周后,测量大鼠双肾质量,计算肾指数,苏木精-伊红(hematoxylin-eosin,HE)染色观察大鼠肾脏病理变化,全自动生化分析仪检测大鼠24 h尿蛋白及血清中血肌酐(Serum creatinine,Scr)、尿素氮(Blood Urea Nitrogen,BUN)水平,按照试剂盒说明书分别检测大鼠血清中空腹血糖(fasting blood glucose, FBG)、甘油三酯(Triglyceride, TG)、胆固醇(Cholesterol, CHO)、白介素-6(Interleukin-6, IL-6)、肿瘤坏死因子-α(tumor necrosis factor α, TNF-α)水平,免疫组化法检测大鼠肾组织中转化生长因子β1(transforming growth factor beta 1,TGF-β1)、细胞间粘附分子-1(intercellular cell adhesion molecule-1,ICAM-1)、血管粘附分子-1(vascular cell adhesion molecule-1,VCAM-1)蛋白的表达。结果 给予糖尿病大鼠EGCG 12周,10、20 mg·kg-1·d-1的EGCG可使其饮水量降低(P < 0.01)、肾小管上皮细胞空泡样病变有所改善;40 mg·kg-1·d-1的EGCG可使其体重升高(P < 0.01),饮水量、摄食量降低(P < 0.01)、肾小管上皮细胞空泡样病变明显得到改善;20、40 mg·kg-1·d-1的EGCG可使其血清中Scr下降(P < 0.01);10、20、40 mg·kg-1·d-1的EGCG可使糖尿病大鼠血清中FBG、TG和CHO水平降低(P < 0.01),肾重和肾脏指数下降(P < 0.01),24 h尿蛋白、血清中BUN、IL-6和TNF-α水平下降(P < 0.05),肾组织中TGF-β1、ICAM-1蛋白表达下降(P < 0.01)。结论 EGCG对糖尿病大鼠肾脏损伤具有改善作用,其机制可能与抑制致纤维化因子TGF-β1、促炎因子IL-6、TNF-α及粘附因子ICAM-1有关。
Abstract
Objective To study the protective effect and mechanism of (-)-Epigallocatechin gallate (EGCG) on kidney of diabetic rats induced by streptozocin (STZ). Methods 30 male SD rats were randomly divided into normal control group, model group, EGCG groups (10, 20, 40 mg·kg-1·d-1). The diabetic rat model was established by intraabdominal injection of STZ (65 mg·kg-1) into the rats in the latter four groups. From the first day after the model was established successfully, the EGCG groups were administered intragastrically according to the above doses, and both the normal control group and the model group were administered intragastrically according to 10 mL·kg-1·d-1 steamed water. With medication, the body weight, drinking water and food intake of the rats in each group were measured at the 6th and 12th week. After 12 weeks, the quality of the two kidneys was measured and the kidney index was calculated. The pathological changes of kidney were observed by hematoxylin-eosin (HE)staining. The levels of Serum creatinine (Scr)and blood urea nitrogen (BUN) in serum and 24 h urinary protein were measured by automatic biochemical analyzer. The levels of fasting blood glucose (FBG), Triglyceride (TG), Cholesterol (CHO), Interleukin-6(IL-6)and tumor necrosis factor α( TNF-α) in serum were detected bytheir respective kit instructions (ELISA). The expressions of transforming growth factor beta 1(TGF-β1), intercellular cell adhesion molecule-1(ICAM-1)and vascular cell adhesion molecule-1(VCAM-1)proteins in renal tissue were detected by immunohistochemistry. Results After diabetic rats were administered intragastrically for 12 weeks, EGCG of 10 and 20 mg·kg-1·d-1 could decrease their water intake (P < 0.01), improve their vacuolar lesion of renal tubular epithelial cells; EGCG of 40 mg·kg-1·d-1 could increase their body weight (P < 0.01), decrease their water intake and food intake (P < 0.01),and significantly improve vacuolar lesions of renal tubular epithelial cells; EGCG of 20 and 40 mg·kg-1·d-1 could decrease the level of Scr in serum(P < 0.01);EGCG of 10, 20 and 40 mg·kg-1·d-1 could decrease the levels of FBG, TG and CHO in serum (P < 0.01), reduce renal weight and renal index (P < 0.01),lower 24 h urinary protein, the levels of BUN, IL-6 and TNF-α in serum (P < 0.05),and inhibit the expression of TGF-β1 and ICAM-1 protein in renal tissue (P < 0.01). Conclusion EGCG could improve the renal damage of diabetic rats by inhibiting the fibrogenic factor TGF-β1, the proinflammatory factor IL-6, TNF-α and the adhesion factor ICAM-1.
关键词
EGCG /
糖尿病肾病 /
TGF-β1 /
促炎因子 /
粘附因子
{{custom_keyword}} /
Key words
EGCG /
Diabetic nephropathy /
TGF-β1 /
proinflammatory factor /
adhesion factor
{{custom_keyword}} /
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] SAEEDI P, SALPEA P, KARURANGA S, et al. Mortality attributable to diabetes in 20-79 years old adults, 2019 estimates: results from the international diabetes federation diabetes atlas[J]. Diabetes Research and clinical Practice,2020(162):108086.
[2] 曾钢,丁群芳.糖尿病微血管并发症及其相关分子机制研究进展[J]. 海南医学院学报,2019,25(1):77-80.
ZENG G, DING Q F. Advances in diabetic microvascular complications and related molecular mechanisms[J]. Journal of Hainan Medical University,2019,25(1):77-80.
[3] ZOJA C, XINARIS C, MACCONI D. Diabetic nephropathy: novel molecular mechanisms and therapeutic targets[J].Front Pharmacol,2020(11):586892.
[4] ALICIC R Z, ROONEY M T, TUTTLE K R. Diabetic kidney disease: challenges, progress, and possibilities[J]. Clin J Am Soc Nephrol,2017(12):2032-2045.
[5] RAYEGO M S, MORGADO P J L, OPAZO R L, et al. Pathogenic pathways and therapeutic approaches targeting inflammation in diabetic nephropathy[J]. Int J Mol Sci,2020, 21(11):3798.
[6] ZHAO J, CHEN J, LI Y Y, et al. Bruton’s tyrosine kinase regulates macrophage?induced inflammation in the diabetic kidney via NLRP3 inflammasome activation[J].Int J Mol Med,2021,48(3):1-12.
[7] ASFOUR M H, SALAMA A A, MOHEN A M. Fabrication of all-trans retinoic acid loaded chitosan/tripolyphosphate lipid hybrid nanoparticles as a novel oral delivery approach for management of diabetic nephropathy in rats[J]. J Pharm Sci,2021,110(9): 3208-3220.
[8] GU Y Y, LIU X S, HUANG X R, et al. Diverse role of TGF-β in kidney disease[J].Front Cell Dev Biol,2020(8):123.
[9] 周静鑫,刘铜华,吴丽丽.中草药干预糖尿病肾病炎症的作用[J].生物化学与生物物理进展,2020,47(8):818-834.
ZHOU J X, LIU T H, WU L L. Regulation of chinese herbal medicines in inflammation of diabetic nephropathy[J]. Acta Biochimica et Biophysica Sinica,2020,47(8):818-834.
[10] SUN W, LIU X, ZHANG H, et al. Epigallocatechin gallate upregulates NRF2 to prevent diabetic nephropathy via disabling KEAP1[J]. Free Radical Biology and Medicine,2017(108):840-857.
[11] HAYASHI D,WANG L,UEDA S, et al. The mechanisms of ameliorating effect of a green tea polyphenol on diabetic nephropathy based on diacylglycerol kinaseα[J].Sci Rep, 2020(10):11790.
[12] MOHAN T, NARASIMHAN K K S, RAVI D B, et al. Role of Nrf2 dysfunction in the pathogenesis of diabetic nephropathy: Therapeutic prospect of epigallocatechin-3-gallate[J]. Free Radic Biol Med,2020(160):227-238.
[13] YAMABE N, YOKOZAWA T, OYA T, et al. Therapeutic potential of (-)-epigallocatechin 3-O-gallate on renal damage in diabetic nephropathy model rats[J]. Journal of Pharmacology and Experimental Therapeutics,2006,319(1):228-236.
[14] GARUD M S, KULKARNI Y A. Gallic acid attenuates type I diabetic nephropathy in rats[J]. Chemico-Biological Interactions,2018(282):69-76.
[15] GROSSO G, STEPANIAK U, MICEK A, et al. Dietary polyphenol intake and risk of type 2 diabetes in the polish arm of the health, alcohol and psychosocial factors in eastern europe (HAPIEE) study[J]. Br J Nutr,2017,118(1):60-68.
[16] BOULMOKH Y, BELGUIDOUM K, MEDDOUR F, et al. Investigation of antioxidant activity of epigallocatechin gallate and epicatechin as compared to resveratrol and ascorbic acid: experimental and theoretical insights[J]. Structural Chemistry,2021(7):1-17.
[17] LIANG O D,KLEIBRINK B E, SCHUETTE N K, et al. Green tea epigallo-catechin-galleate ameliorates the development of obliterative airway disease[J]. Exp Lung Res,2011,37(7):435-444.
[18] WANG M,ZHONG H,ZHANG X, et al. EGCG promotes PRKCA expression to alleviate LPS-induced acute lung injury and inflammatory response[J]. Sci Rep,2021,11(1):11014.
[19] BAEK C H, KIM H, MOON S Y, et al. Epigallocatechin-3-gallate downregulates lipopolysaccharide signaling in human aortic endothelial cells by inducing ectodomain shedding of TLR4[J]. Eur J Pharmacol,2019(863):172692.
[20] XIE H, SUN J, CHEN Y, et al. (-)-Epigallocatechin-3-gallate protects against uric acid-induced endothelial dysfunction in human umbilical vein endothelial cells[J]. Pharmacognosy Magazine,2019,15(63):487.
[21] LAKSHMI S P, REDDY A T, KODIDHELA L D, et al. The tea catechin epigallocatechin gallate inhibits NF-κB-mediated transcriptional activation by covalent modification[J]. Archives of Biochemistry and Biophysics,2020(695):108620.
[22] 李雪松,方明儒,江伟杰,等.七芪地黄丸对糖尿病肾病大鼠的保护作用及机制研究[J]. 中药药理与临床,2019,35(3):127-133.
LI X S, FANG M R,JIANG W J, et al. Pharmacology and clinics of chinese materia medicaprotective effect and mechanism of Qiqi Dihuang Pill on diabetic nephropathy rats[J]. Pharmacology and Clinics of Chinese Materia Medica, 2019,35(3):127-133.
[23] KAJITAN N, SHIKATA K, NAKAMURA A, et al. Microinflammation is a common risk factor for progression of nephropathy and atherosclerosis in Japanese patients with type 2 diabetes[J]. Diabetes Research and Clinical Practice,2010,88(2):171-176.
[24] OMODANISI E, ABOUA Y, OGUNTIBEJU O. Assessment of the anti-hyperglycaemic, anti-inflammatory and antioxidant activities of the methanol extract of moringa oleifera in diabetes-induced nephrotoxic male wistar rats[J]. Molecules,2017,22(4): 439.
[25] WAN R J, LI Y H. MicroRNA146a/NAPDH oxidase4 decreases reactive oxygen species generation and inflammation in a diabetic nephropathy model[J]. Molecular Medicine Reports,2018,17(3):4759-4766.
[26] NAWAZ S S, JOY S S, AL F Y, et al. Potential role of serum fetuin-a in relation with pro-inflammatory, chemokine and adhesion molecules in diabetic kidney disease: a case-control study[J]. Molecular Biology Reports,2019,46(1):1239-1246.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
基金
辽宁省教育厅自然科学基金项目(JYTFW201915)
{{custom_fund}}
PDF(8834 KB)
