Effect of adiponectin in the treatment of nephrotic syndrome rats undergoing adipose stem cell transplantation

doi:10.3969/j.issn.2095-4344.0519

Abstract

Abstract:

BACKGROUND: Adiponectin can regulate glucose and lipid metabolism in the body and protect body tissues and organs. Therefore, we combine it with stem cell transplantation for the treatment of nephrotic syndrome.
OBJECTIVE: To investigate the therapeutic effect of adiponectin combined with adipose stem cell transplantation in the rats with nephrotic syndrome.
METHODS: Rat adipose stem cells were resuscitated and cultured in vitro, and labeled using CD-Dil prior to the transplantation. Eighty Sprague-Dawley rats were randomly assigned into a normal control group, a model group, an adipose stem cell transplantation group, and a combined group (adiponectin and adipose stem cell transplantation), with 20 rats in each group. Rats in the normal control group received no treatment. The remaining rats in the latter three groups were given a single intravenous injection of adriamycin (6 mg/kg) to prepare animal models of nephrotic syndrome, followed by a tail vein injection of normal saline, 2×10⁶/L adipose stem cell suspension, and 1 μg/kg adiponectin plus 2×10⁶/L adipose stem cell suspension, respectively. Administration of adiponectin in the combined group was given once a day, for 3 consecutive days.
RESULTS AND CONCLUSION: (1) Compared with the normal control group, higher levels of 24-hour urinary protein, blood cholesterol and urea nitrogen as well as lower serum albumin level were observed in the remaining three groups at 28 days after modeling. Compared with the model group, the levels of 24-hour urinary protein, blood cholesterol and urea nitrogen in the adipose stem cell transplantation group were decreased, while the level of serum albumin was increased. Compared with the adipose stem cell transplantation group, the levels of 24-hour urinary protein, blood cholesterol and urea nitrogen in the combined group were decreased, while the level of serum albumin was increased. (2) At 28 days after modeling, severest renal damage appeared in the model group, while the damage was reduced in the adipose stem cell transplantation group and considerably relieved in the combined group accompanied by improved inflammation and edema. (3) The number of CM-Dil positive cells in renal tissues was highest in the combined group followed by the adipose stem cell transplantation group, but no positive cells were detected in the normal control and model groups. Our experimental findings reveal that adipose-derived stem cell transplantation has a certain therapeutic effect on nephrotic syndrome, and adiponectin can enhance this therapeutic effect, by improving renal function and effectively inhibiting the pathological changes of renal tissues.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Nephrosis, Adiponectin, Stem Cells, Tissue Engineering

CLC Number:

R394.2

Liu Qian, Zhang Yu-jing, Liu Jing, Wang Jing, Xie Jin. Effect of adiponectin in the treatment of nephrotic syndrome rats undergoing adipose stem cell transplantation[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(13): 2109-2113.

Figures/Tables 1

2.1 脂肪干细胞培养情况脂肪干细胞在培养皿中传代至第4代时，约2 d即可达到80%-90%以上的融合。7 d后，大部分细胞呈典型成纤维细胞样形态，分布均匀，形态均一，呈旋涡状生长，杂细胞少见或几乎不可见，镜下可见单个细胞表面光滑，胞浆丰富，核仁及结构清晰明显(图1A)。CM-Dil标记的脂肪干细胞在荧光显微镜下呈红色荧光(图1B)。 2.2 实验动物数量变化 80只大鼠，营养状况良好，建造肾病综合征模型顺利，无死亡例数，进行后续实验时也无一例死亡，所有大鼠均进入结果分析。 2.3 大鼠血、尿指标变化与对照组相比，另3组大鼠24 h尿蛋白、血胆固醇和尿素氮水平均上升，而血白蛋白水平下降(P < 0.05)；与肾病综合征模型组相比，脂肪干细胞移植组大鼠24 h尿蛋白、血胆固醇和尿素氮值降低，血白蛋白升高(P < 0.05)；与脂肪干细胞移植组相比，脂联素联合脂肪干细胞移植组24 h尿蛋白、血胆固醇和尿素氮值降低，血白蛋白升高(P < 0.01)，见表1。 2.4 肾组织病理学变化光镜下，对照组大鼠肾小球滤过屏障清晰，厚度均匀，足细胞突起整齐地贴附在基膜上，有孔毛细血管内皮清楚。肾病综合征模型组肾小球体积增大，毛细血管袢开放不良，系膜细胞增生系膜基质增多，系膜区增宽。脂肪干细胞移植组以上病理变化较肾病综合征模型组减轻，脂联素联合脂肪干细胞移植组大鼠肾脏病理形态学变化明显减轻(图2)。 2.5 肾组织中脂肪干细胞的存活情况荧光显微镜下观察可见对照组和肾病综合征模型组大鼠肾组织中没有CM-Dil阳性细胞，而脂肪干细胞移植后，大鼠肾组织出现大量的CM-Dil阳性细胞，且联合脂联素治疗的大鼠肾组织CM-Dil阳性细胞多于脂肪干细胞移植组(图3)。"

References

[1] Gibson KL, Glenn D, Ferris ME. Back to the future: therapies for idiopathic nephrotic syndrome. Blood Purif. 2015;39(1-3): 105-109.

[2] Bierzynska A, Saleem M. Recent advances in understanding and treating nephrotic syndrome. F1000Res. 2017;6:121.

[3] Colucci M, Corpetti G, Emma F, et al. Immunology of idiopathic nephrotic syndrome. Pediatr Nephrol. 2018;33(4):573-584.

[4] Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng. 2001;7(2):211-228.

[5] 陈红梅,沈英杰,房艳.ADSCs对糖尿病肾病大鼠肾纤维化的抑制作用[J].解剖科学进展,2015,21(3):236-240.

[6] Sheashaa H, Lotfy A, Elhusseini F, et al. Protective effect of adipose-derived mesenchymal stem cells against acute kidney injury induced by ischemia-reperfusion in Sprague-Dawley rats. Exp Ther Med. 2016;11(5):1573-1580.

[7] 郭志伯,张晨洁,马李娜,等.脂肪间充质干细胞移植联合人工虫草多糖改善肾病综合征的高凝血状态及肾功能[J].中国组织工程研究, 2016,20(50):7549-7556.

[8] 郭佳,肖传实,白瑞,等.脂联素对大鼠缺血/再灌注心肌细胞凋亡及相关蛋白表达的影响[J].中国药理学通报,2012,28(7):930-933.

[9] Opatrilova R, Caprnda M2, Kubatka P3, et al. Adipokines in neurovascular diseases. Biomed Pharmacother. 2018;98: 424-432.

[10] Schmued LC, Stowers CC, Scallet AC, et al. Fluoro-Jade C results in ultra high resolution and contrast labeling of degenerating neurons. Brain Res. 2005;1035(1):24-31.

[11] Kuroda K, Kabata T, Hayashi K, et al. The paracrine effect of adipose-derived stem cells inhibits osteoarthritis progression. BMC Musculoskelet Disord. 2015;16:236.

[12] Letra L, Rodrigues T, Matafome P, et al. Adiponectin and sporadic Alzheimer's disease: Clinical and molecular links. Front Neuroendocrinol. 2017. doi: 10.1016/j.yfrne.2017.10.002.

[13] Adolph TE, Grander C, Grabherr F, et al. Adipokines and Non-Alcoholic Fatty Liver Disease: Multiple Interactions. Int J Mol Sci. 2017;18(8): E1649.

[14] 朱慧娟,任嘉庆,于双玉.肥胖儿童血浆脂联素水平及对肾损伤作用[J].中国公共卫生,2010,26(1):25-26.

[15] 桂定坤.黄芪水提物促肾脏水钠排泄及肾脏保护作用的细胞分子机制研究[D].上海:复旦大学,2007.

[16] 许文芳,柳红梅.应用磺基水杨酸-硫酸钠比浊法在自动化分析仪上测定尿蛋白[J].检验医学,2007,22(1):23-24.

[17] 张茵,银永革.肾病综合征的治疗进展[J].医学综述,2014,20(2): 260-262.

[18] 樊威伟.肾病综合征与相关细胞因子的研究及治疗进展[J].湖南中医杂志,2014,30(2):150-152.

[19] 何开英.肾病综合征中医治疗研究进展[J].亚太传统医药,2016, 12(1):44-45.

[20] 李可.脂肪组织对其他组织器官的调控作用[J].中国高新技术企业, 2017(8):88-89.

[21] Sweiss N, Sharma K. Adiponectin effects on the kidney. Best Pract Res Clin Endocrinol Metab. 2014;28(1):71-79.

[22] 张同梅.脂联素与多种疾病关系的研究进展[J].实用心脑肺血管病杂志,2015,23(2):4-7.

[23] Yano Y, Hoshide S, Ishikawa J, et al. Differential impacts of adiponectin on low-grade albuminuria between obese and nonobese persons without diabetes. J Clin Hypertens (Greenwich). 2007;9(10):775-782.

[24] Nolan E, O'Meara YM, Godson C. Lipid mediators of inflammation in obesity-related glomerulopathy. Nephrol Dial Transplant. 2013;28 Suppl 4:iv22-29.

[25] 侯洁,许芳,许颖.脂联素与慢性肾脏病的研究进展[J].中华临床医师杂志(电子版),2016,10(11):1595-1600.

[26] Perri A, Vizza D, Lupinacci S, et al. Adiponectin secreted by tubular renal cells during LPS exposure worsens the cellular inflammatory damage. J Nephrol. 2016;29(2):185-194.

[27] O'Halloran N, Courtney D, Kerin MJ, et al. Adipose-Derived Stem Cells in Novel Approaches to Breast Reconstruction: Their Suitability for Tissue Engineering and Oncological Safety. Breast Cancer (Auckl). 2017;11:1178223417726777.

[28] Tabatabaei Qomi R, Sheykhhasan M. Adipose-derived stromal cell in regenerative medicine: A review. World J Stem Cells. 2017;9(8):107-117.

[29] Bora P, Majumdar AS. Adipose tissue-derived stromal vascular fraction in regenerative medicine: a brief review on biology and translation. Stem Cell Res Ther. 2017;8(1):145.

[30] 祁美武,周晓.脂肪干细胞的研究进展[J].中南医学科学杂志,2015, 43(3):342-345.

[31] 孙惠君,李博,陈书强,等.脂肪干细胞的研究进展[J].现代生物医学进展,2016,16(12):2380-2383.

[32] Lim S, Cho H, Lee E, et al. Osteogenic stimulation of human adipose-derived stem cells by pre-treatment with fibroblast growth factor 2. Cell Tissue Res. 2016;364(1):137-147.

[33] Brown CF, Yan J, Han TT, et al. Effect of decellularized adipose tissue particle size and cell density on adipose-derived stem cell proliferation and adipogenic differentiation in composite methacrylated chondroitin sulphate hydrogels. Biomed Mater. 2015;10(4):045010.

[34] 徐挺,吴慧玲,胡俊彪,等.采用动物模型探究脂肪干细胞促进肾积水功能恢复的实验研究[J].中国高等医学教育,2016(6):127-135.

[35] 刘少鹏.经肾动脉移植脂肪干细胞对大鼠急性缺血性肾损伤的治疗作用研究[D].上海:复旦大学,2014.

[36] Burgos-Silva M, Semedo-Kuriki P, Donizetti-Oliveira C, et al. Adipose Tissue-Derived Stem Cells Reduce Acute and Chronic Kidney Damage in Mice. PLoS One. 2015;10(11):e0142183.

[37] Furuichi K, Shintani H, Sakai Y, et al. Effects of adipose-derived mesenchymal cells on ischemia-reperfusion injury in kidney. Clin Exp Nephrol. 2012;16(5):679-689.

[38] Rivera-Valdés JJ, García-Bañuelos J, Salazar-Montes A, et al. Human adipose derived stem cells regress fibrosis in a chronic renal fibrotic model induced by adenine. PLoS One. 2017;12(12): e0187907.

[39] Xue A, Niu G, Chen Y, et al. Recellularization of well-preserved decellularized kidney scaffold using adipose tissue-derived stem cells. J Biomed Mater Res A. 2018;106(3):805-814.

[40] Sheashaa H, Lotfy A, Elhusseini F, et al. Protective effect of adipose-derived mesenchymal stem cells against acute kidney injury induced by ischemia-reperfusion in Sprague-Dawley rats. Exp Ther Med. 2016;11(5):1573-1580.

[41] Wang YL, Li G, Zou XF, et al. Effect of autologous adipose-derived stem cells in renal cold ischemia and reperfusion injury. Transplant Proc. 2013;45(9):3198-3202.