姜黄素对5/6肾结扎诱导的慢性肾病小鼠模型肾纤维化的保护作用
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摘要
目的研究姜黄素对慢性肾病(CKD)肾纤维化的保护作用,并探讨可能作用机制。方法 30只C57BL/6 J小鼠随机分成假手术对照组(NC)、5/6肾结扎模型组(LIG)和姜黄素治疗组(LIG+CUR),每组10只,按照改良的5/6肾结扎方法制作CKD动物模型,姜黄素组小鼠给予含姜黄素饲料(每日100 mg/kg),其余组给予正常饲料,三月后牺牲小鼠,检测纤维化指标α-SMA及参与慢性肾病纤维化的Hippo通路转录激活子Yap。结果肾功能检测结果显示,与假手术组相比,结扎组中的尿素氮(BUN)、血肌酐(Scr)显著增高,给予姜黄素可以有效保护肾功能受损;H&E、Masson和免疫组化结果均显示,结扎组小鼠的肾出现明显肾小管病变和一定程度的纤维化改变,给予姜黄素后纤维化程度显著减轻(P<0.05);而Yap蛋白在造模后mRNA和蛋白水平均出现升高,姜黄素处理后则显著下降(P<0.05)。结论姜黄素可有效改善5/6肾结扎诱导的慢性肾病病症和减轻纤维化病变,机制研究结果初步提示可能与降低Hippo信号通路中的Yap表达有关。
Objective To study the protective effect of curcumin on renal fibrosis in chronic kidney disease(CKD) and explore its possible mechanism of action. Methods Thirty C57 BL/6 J mice were randomly divided into a sham-operated control group(NC), 5/6 kidney ligation model group(LIG) and curcumin treatment group(LIG + CUR), with 10 mice in each group. The CKD animal model was prepared according to the modified 5/6 kidney ligation method. The curcumin group was given a curcumin-containing diet(100 mg/(kg·d) and the other groups were given normal feed. The mice were euthanized after 3 months. The levels of the fibrosis index α-SMA and the Hippo pathway transcriptional activator Yap involved in fibrosis in CKD were determined. Results Renal function tests showed that urea nitrogen(BUN) and serum creatinine(Scr) were significantly increased in the LIG group compared with the NC group, and that the administration of curcumin effectively protected kidney function. H&E, Masson and immunohistochemical staining showed that the LIG kidneys had obvious tubular lesions and some fibrotic changes, and that the degree of fibrosis was significantly reduced after the administration of curcumin(P<0.05). The mRNA and protein levels of Yap increased after modeling, and decreased significantly after curcumin treatment(P<0.05). Conclusions Curcumin can significantly ameliorate the mouse CKD induced by 5/6 kidney ligation, and its mechanism may be related to a decrease of Yap in the Hippo signaling pathway.
Objective To study the protective effect of curcumin on renal fibrosis in chronic kidney disease(CKD) and explore its possible mechanism of action. Methods Thirty C57 BL/6 J mice were randomly divided into a sham-operated control group(NC), 5/6 kidney ligation model group(LIG) and curcumin treatment group(LIG + CUR), with 10 mice in each group. The CKD animal model was prepared according to the modified 5/6 kidney ligation method. The curcumin group was given a curcumin-containing diet(100 mg/(kg·d) and the other groups were given normal feed. The mice were euthanized after 3 months. The levels of the fibrosis index α-SMA and the Hippo pathway transcriptional activator Yap involved in fibrosis in CKD were determined. Results Renal function tests showed that urea nitrogen(BUN) and serum creatinine(Scr) were significantly increased in the LIG group compared with the NC group, and that the administration of curcumin effectively protected kidney function. H&E, Masson and immunohistochemical staining showed that the LIG kidneys had obvious tubular lesions and some fibrotic changes, and that the degree of fibrosis was significantly reduced after the administration of curcumin(P<0.05). The mRNA and protein levels of Yap increased after modeling, and decreased significantly after curcumin treatment(P<0.05). Conclusions Curcumin can significantly ameliorate the mouse CKD induced by 5/6 kidney ligation, and its mechanism may be related to a decrease of Yap in the Hippo signaling pathway.
引文
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[11] Sugiura H, Matsushita A, Futaya M, et al. Fibroblast growth factor 23 is upregulated in the kidney in a chronic kidney disease rat model [J]. PLoS One, 2018, 13(3): e0191706.
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[13] Wang XT, Sun XJ, Li C, et al. Establishing a cell-based high-content screening assay for TCM compounds with anti-renal fibrosis effects [J]. Evid Based Complement Alternat Med, 2018, 7942614.
[14] Ali BH, Al-Salam S, Al Suleimani Y, et al. Curcumin ameliorates kidney function and oxidative stress in experimental chronic kidney disease [J]. Basic Clin Pharmacol Toxicol, 2018, 122(1): 65-73.
[15] Liu F, Lagares D, Choi KM, et al. Mechanosignaling through YAP and TAZ drives fibroblast activation and fibrosis [J]. Am J Physiol Lung Cell Mol Physiol, 2015, 308(4): L344-57.
[16] Mitani A, Nagase T, Fukuchi K, et al. Transcriptional coactivator with PDZ-binding motif is essential for normal alveolarization in mice [J]. Am J Respir Crit Care Med, 2009, 180(4): 326-338.
[17] Zeng F, Miyazawa T, Kloepfer LA, et al. ErbB4 deletion accelerates renal fibrosis following renal injury [J]. Am J Physiol Renal Physiol, 2018, 314(5): F773-F787.
[18] Anorga S, Overstreet JM, Falke LL, et al. Deregulation of Hippo-TAZ pathway during renal injury confers a fibrotic maladaptive phenotype [J]. FASEB J, 2018, 32(5): 2644-2657.
[2] Zhong W, Qian K, Xiong J, et al. Curcumin alleviates lipopolysaccharide induced sepsis and liver failure by suppression of oxidative stress-related inflammation via PI3K/AKT and NF-κB related signaling [J]. Biomed Pharmacother, 2016, 83: 302-313.
[3] Liu Y, Wu YM, Yu Y, et al. Curcumin and resveratrol in combination modulate drug-metabolizing enzymes as well as antioxidant indices during lung carcinogenesis in mice [J]. Hum Exp Toxicol, 2015, 34(6): 620-627.
[4] Zhao XA, Chen G, Liu Y, et al. Curcumin reduces Ly6C monocyte infiltration to protect against liver fibrosis by inhibiting Kupffer cells activation to reduce chemokines secretion [J]. Biomed Pharmacother, 2018, 106: 868-878.
[5] Lau WL, Khazaeli M, Savoj J, et al. Dietary tetrahydrocurcumin reduces renal fibrosis and cardiac hypertrophy in 5/6 nephrectomized rats [J]. Pharmacol Res Perspect, 2018, 6(2): e00385.
[6] Aparicio-Trejo OE, Tapia E, Molina-Jijón E, et al. Curcumin prevents mitochondrial dynamics disturbances in early 5/6 nephrectomy: Relation to oxidative stress and mitochondrial bioenergetics [J]. Biofactors, 2017, 43(2): 293-310.
[7] Wang X, Chaudhry MA, Nie Y, et al. A mouse 5/6th nephrectomy model that induces experimental uremic cardiomyopathy [J]. J Vis Exp, 2017, 7(129): 1-6.
[8] Fan Y, Chen H, Peng H, et al. Molecular mechanisms of curcumin renoprotection in experimental acute renal injury [J]. Front Pharmacol, 2017, 8: 912.
[9] 宋娟,孙燕妮,秦熠,等.侧脑室注入网膜素对小鼠摄食量及其下丘脑组织NPY、AGRP、CRH、POMC mRNA表达的影响 [J]. 山东医药, 2015, 55(31): 37-38.Song J, Sun YN, Qin Y, et al. Effects of injecting retinal into lateral ventricle on the food intake of mice and the expression of NPY, AGRP, CRH and POMC mRNA in hypothalamus [J]. Shandong Med, 2015, 55(31): 37-38.
[10] 黄统生, 郭赟, 杨陈, 等. 反复注射顺铂诱导小鼠慢性肾功能不全模型的建立 [J]. 中国实验动物学报,2018, 26(1): 20-28.Huang TS, Guo Y, Yang C, et al. Establishment of a model of chronic renal insufficiency induced by repeated injection of cisplatin in mice [J]. Acta Lab Anim Sci Sin, 2018, 26(1): 20-28.
[11] Sugiura H, Matsushita A, Futaya M, et al. Fibroblast growth factor 23 is upregulated in the kidney in a chronic kidney disease rat model [J]. PLoS One, 2018, 13(3): e0191706.
[12] Zou H, Zhao X, Sun N, et al. Effect of chronic kidney disease on the healing of titanium implants [J]. Bone, 2013, 56(2): 410-415.
[13] Wang XT, Sun XJ, Li C, et al. Establishing a cell-based high-content screening assay for TCM compounds with anti-renal fibrosis effects [J]. Evid Based Complement Alternat Med, 2018, 7942614.
[14] Ali BH, Al-Salam S, Al Suleimani Y, et al. Curcumin ameliorates kidney function and oxidative stress in experimental chronic kidney disease [J]. Basic Clin Pharmacol Toxicol, 2018, 122(1): 65-73.
[15] Liu F, Lagares D, Choi KM, et al. Mechanosignaling through YAP and TAZ drives fibroblast activation and fibrosis [J]. Am J Physiol Lung Cell Mol Physiol, 2015, 308(4): L344-57.
[16] Mitani A, Nagase T, Fukuchi K, et al. Transcriptional coactivator with PDZ-binding motif is essential for normal alveolarization in mice [J]. Am J Respir Crit Care Med, 2009, 180(4): 326-338.
[17] Zeng F, Miyazawa T, Kloepfer LA, et al. ErbB4 deletion accelerates renal fibrosis following renal injury [J]. Am J Physiol Renal Physiol, 2018, 314(5): F773-F787.
[18] Anorga S, Overstreet JM, Falke LL, et al. Deregulation of Hippo-TAZ pathway during renal injury confers a fibrotic maladaptive phenotype [J]. FASEB J, 2018, 32(5): 2644-2657.