摘要
目的:探讨丹酚酸B调控自噬改善肾纤维化的作用机制。方法:60只小鼠随机分为6组:假手术组、模型组、醋酸泼尼松(20 mg/kg)阳性对照组及丹酚酸B低、中、高剂量(50、100、200 mg/kg)组,除假手术组外各组均采用单侧输尿管梗阻(UUO)的方法建立小鼠肾纤维化模型。自造模当天各组给予相应药物,假手术组和模型组给予等体积生理盐水灌胃。术后第7天处死小鼠取肾取血,测定血清肌酐和尿素氮,HE染色和Masson染色观察肾脏组织病理变化及纤维化程度,免疫组化法观察肾间质Vimentin的表达,蛋白免疫印记法检测α-SMA及自噬相关蛋白LC3-Ⅱ、Beclin1、mTOR的表达。结果:与模型比较,丹酚酸B可降低UUO小鼠血肌酐和尿素氮水平,HE染色和Masson染色结果显示肾组织病理损伤减轻,胶原纤维沉积减少。免疫组织化学法和Western Blot结果表明丹酚酸B可使UUO小鼠肾组织中纤维化标记蛋白Vimentin、α-SMA表达下调,而自噬标记蛋白LC3-Ⅱ、Beclin1表达上调,且mTOR表达下调。结论:丹酚酸B能改善UUO模型小鼠肾纤维化,其作用机制可能与激活肾组织自噬有关。
引文
[1] Genovese,F,Manresa AA,Leeming DJ,et al.The extracellular matrix in the kidney:a source of novel non-invasive biomarkers of kidney fibrosis[J].Fibrogenesis Tissue Repair,2014,7(1):4-14.
[2] Wang QL,Tao YY,Yuan JL,et al.Salvianolic acid B prevents epithelial-to-mesenchymal transition through the TGF-beta1 signal transduction pathway in vivo and in vitro[J].BMC Cell Biol,2010,11(1):31-40.
[3] 蔡洲,李道,吴际,等.丹酚酸B对肾间质纤维化大鼠的肾保护作用及其机制[J].山东医药,2017,57(20):34-36.
[4] Havasi A,Zheng D.Autophagy and Tubular Cell Death in the Kidney[J].Semin Nephrol,2016,36(3):174-88.
[5] He L,Livingston MJ,Zheng D.Autophagy in Acute Kidney Injury and Repair[J].Nephron Clin Pract,2014,127(1-4):56-60.
[6] Kramann R,Dirocco DP,Maarouf OH,et al.Matrix-producing cells in chronic kidney disease:origin,regulation,and activation[J].Curr Pathobiol Rep,2013,1(4):301-311.
[7] 于金玲.丹酚酸B药理作用研究进展[J].天津药学,2012,24(3):64-68.
[8] Kim WY,Nam SA,Song HC,et al.The role of autophagy in unilateral ureteral obstruction rat model[J].Nephrology,2012,17(2):148-159.
[9] Wang Z,Divanyan A,Jourd′heuil F,et al.Vimentin expression is required for the development of EMT-related renal fibrosis following Unilateral Ureteral Obstruction in mice[J].Am J Physiol Renal Physiol,2018,315(4):F769-F780.
[10] 陆海英,张悦,刘煜敏,等.α-平滑肌肌动蛋白和E-钙依赖性黏附素在肾间质纤维化大鼠肾组织中的表达[J].中国病理生理杂志,2009,25(4):795-798.
[11] 黄锐.Sirt1调控核内LC3脱乙酰化在自噬中的作用研究[D].杭州:浙江大学,2013.
[12] 黄蓉,韦曦.自噬效应蛋白Beclin1的研究进展[J].中外医学研究,2011,9(28):159-161.
[13] Xu Y,Yang S,Huang J,et al.TGF-betal1 in duces autophagy and promotes apoptosis in renal tubular epithelial cells[J].Int J Mol Med,2012,29(5):781-790.