MicroRNA expression profiling in male and female model mice after renal ischemia-reperfusion injury

doi:10.3969/j.issn.2095-4344.2015.05.020

Abstract

Abstract:

BACKGROUND: Renal ischemia-reperfusion injury has been shown to exhibit gender difference, but its precise mechanisms deserve further investigations.
OBJECTIVE: To investigate the differential expression of microRNAs in the kidney between female and male mice in order to study the effects and mechanisms of microRNA in pathogenesis of ischemia-reperfusion injury between different genders.
METHODS: Male and female mice received kidney ischemia for 45 minutes and reperfusion injury for 24 hours. Simultaneously, male and female sham surgery groups served as controls. The microRNA gene chip technology was used to detect the differences of microRNA expression in the kidney of male and female mice at 45 minutes after ischemia and 24 hours of reperfusion as well as after sham surgery. The threshold of difference in expression among samples was double.
RESULTS AND CONCLUSION: Five microRNAs were up-regulated between female and male ischemia-reperfusion injury groups. Twenty-nine microRNAs differentially expressed in the female ischemia-reperfusion group and female sham surgery group, including 25 up-regulated microRNAs and 4 down-regulated microRNAs. Thirty-eight microRNAs differentially expressed in male ischemia-reperfusion injury group and male sham surgery group, including 9 up-regulated microRNAs and 29 down-regulated microRNAs. 102 microRNAs differentially expressed in the female sham surgery group and male sham surgery group, including 22 up-regulated microRNAs and 80 down-regulated microRNAs. Results suggested that there was differential expression in microRNAs in the kidney before and after renal ischemia-reperfusion in male and female mice. These differentially expressed microRNAs may be lead to different sensitivity and tolerance to the ischemia-reperfusion injury in the kidney of male and female mice.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

全文链接：

Key words: Tissue Engineering, Kidney, Reperfusion Injury, Microchip Analytical Procedures

CLC Number:

R318

Tang Wei-wei, Xi Xiao-qing, Hu Hong-lin, Huang Ya-wei, Ye Zhen-feng, Chen Ding-yi . MicroRNA expression profiling in male and female model mice after renal ischemia-reperfusion injury[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(5): 772-777.

Figures/Tables 4

2.1 实验动物数量分析纳入BALB/c小鼠80只，建模失败后立即补充试验用BALB/c小鼠，进入MicroRNA芯片结果分析共12只，其中每组各3只。 2.2 血清肌酐及尿素氮检测结果肾脏灌注后24 h，经眼眶内眦静脉丛取血，检测各组血清肌酐(Cr)和尿素氮(BUN)结果发现：本组实验中，雌性假手术组小鼠血清肌酐约15 µmol/L，尿素氮约7.2 mmol/L。小鼠经历45 min肾脏缺血，再灌注后24 h血清肌酐和尿素氮值开始升高，分别达到(20.15±2.31) µmol/L和(9.24±0.63) mmol/L，雌性肾缺血再灌注组与雌性假手术组比较后血清肌酐及血清尿素氮变化无统计学意义，见图1。而雄性假手术组小鼠血清肌酐 14 µmol/L，尿素氮6.8 mmol/L。鼠经历45 min肾脏缺血，再灌注后24 h血清肌酐和尿素氮值开始升高(P < 0.05)，分别达到(84.35±5.26) µmol/L和(27.36±2.15) mmol/L。 2.3 不同性别小鼠肾缺血再灌注损伤后肾组织病理形态各组肾脏经苏木精-伊红染色后在200倍显微镜下病理结果采用Rabb病理评估方法雄性假手术组及雌性假手术组肾组织基本正常，仅建部分肾小管上皮细胞水肿，肾脏病理评分为0分，雄性肾缺血再灌注组可见肾小管上皮细胞坏死、空泡变性以及细胞核浓缩、碎裂、核溶解如图1箭头所示，病理损伤范围约为50%，肾脏病理评分为3分；雌性肾缺血再灌注组肾小管上皮细胞坏死、崩解脱落以及细胞核浓缩、碎裂、核溶解如图，箭头所示。部分肾小管管腔扩张、管腔轮廓模糊，病理损伤范围大于75%，故肾脏病理评分为4分。采用Rabb病理评估方法雄性假手术组及雌性假手术组肾小管细胞无明显变化肾脏病理评分为0分，雄性肾缺血再灌注组可见肾小管细胞大量空泡变性如图1箭头所示，肾脏病理评分为3分；雌性肾缺血再灌注组肾小管细胞出现液化坏死、核固缩如图箭头所示，肾小管形态缺失，故肾脏病理评分为4分。"

References

[1] TanakaR, Tsutsui H, Ohkita M, et al. Sex differences in ischemia/reperfusion-induced acute kidney injury are dependent on the renal sympathetic nervous system. Eur J Pharmacol. 2013;714(1-3):397-404.
[2] 徐青云,邵凤民.性别差异在肾脏缺血再灌注损伤中的作用机制[J].实用诊断与治疗杂志,2007,21(5):362-364
[3] Park KM, Kim JI, Ahn Y, et al. Testosterone is responsible for enhanced susceptibility of males to ischemic renal injury. J Biol Chem. 2004;279(50):52282-52292.
[4] Wei Q, Wang MH, Dong Z, et al. Differential gender differences in ischemic and nephrotoxic acute renal failure. Am J Nephrol. 2005;25(5):491-499.
[5] Bartel B, Bartel DP. MicroRNAs: at the root of plant development? Plant Physiol. 2003;132(2):709-717.
[6] Lim LP, Glasner ME, Yekta S, et al. Vertebrate microRNA genes. Science. 2003;299(5612):1540.
[7] Hwang HW, Mendell JT. MicroRNAs in cell proliferation, cell death, and tumorigenesis.Br J Cancer. 2006;94(6):776-780.
[8] Croce CM, Calin GA. miRNAs, cancer, and stem cell division. Cell. 2005;122(1):6-7.
[9] Ma L, Teruya-Feldstein J, Weinberg RA. Tumour invasion and metastasis initiated by microRNA-10b in breast cancer. Nature. 2007;449(7163):682-688.
[10] Tavazoie SF, Alarcón C, Oskarsson T, et al. Endogenous human microRNAs that suppress breast cancer metastasis. Nature. 2008;451(7175):147-152.
[11] Ren A, Yan X ,Lu H, et al. Antagonism of endothelin-1inhibits hypoxia-induced apoptosis in cardiomyocytes. Can J Physiol Pharmacol. 2008;86:536-540.
[12] Xu C, Lu Y, Pan Z, et al. The muscle-specific microRNAs miR-1 and miR-133 produce opposing effects on apoptosis by targeting HSP60 HSP70 and caspase-9 in cardiomyocytes. J Cell Sci. 2007;120(Pt 17):3045-3052.
[13] The Ministry of Science and Technology of the People’s Republic of China. Guidance Suggestions for the Care and Use of Laboratory Animals. 2006-09-30.
[14] 胡红林,王共先,邹丛.BALB/c小鼠肾缺血再灌注损伤模型的建立与评价[J].中国组织工程研究与临床康复,2011,15(5):870-873.
[15] Rabb H, Mendiola CC, Dietz J, et al. Role of CD11a and CD11b in ischemic acute renal failure in rats.Am J Physiol. 1994;267(6 Pt 2):F1052-F1058.
[16] Wei Q, Wang MH, Dong Z. Differential gender differences in ischemic and nephrotoxic acute renal failure. Am J Nephrol. 2005;25(5):491-499.
[17] Kittikulsuth W, Sullivan JC, Pollock DM. ET-1 actions in the kidney: evidence for sex differences. Pharmacology. 2013; 168: 318-326.
[18] Fukuda K, Yao H, Ibayashi S, et al. Ovariectomy exacerbates and estrogen replacement attenuates photothrombotic focal ischemic brain injury in rats. Stroke. 2000;31:155-160.
[19] Sugden PH, Clerk A. Akt like a woman: gender differences in susceptibility to cardiovascular disease. Circ Res. 2001;88: 975-977.
[20] Muller V. Losonczy G. Heemann U. et al. Sexual dimorphism in renal ischemia-reperfusion injury in rats: possible role of endothelin. Kidney Int. 2002;62:1364-1371.
[21] Park KM, Kim JI, Ahn Y, et al. Testosterone is responsible for enhanced susceptibility ofmales to ischemic renal injury. J Biol Chem. 2004;279(50):52282-52292.
[22] Fekete A, Vannay A, Tulassay T, et al. Sex difference in the alterations of Na(+), K(+)-ATPase following ischemia- reperfusion injury in the rat kidney. J Physiol 2004;555(2): 471-480.
[23] 肖占琴,陶树新,陈阳美. microRNA 在神经系统疾病发病机制中的作用[J].中华神经医学杂志,2011,10(2):204-206.
[24] 雷平,李耀华,杨树源,等. microRNA 在神经系统中的作用研究进展[J].中华神经医学杂志,2010,9(2):203-207.
[25] 谭健,肖健,王志农.MicroRNA在大鼠心肌缺血再灌注损伤中表达谱的变化[J].实用医学杂志,2013,28(2):199-201.
[26] Jia P, Teng J, Zou J, et al. miR-21 contributes to xenon-conferred amelioration of renal ischemia-reperfusion injury in mice.Anesthesiology. 2013;119(3):621-630.
[27] Kaucsár T, Révész C, Godó M, et al. Activation of the miR-17 family and miR-21 during murine kidney ischemia-reperfusion injury. Nucleic Acid Ther. 2013;23(5):344-354.
[28] Saikumar J, Hoffmann D, Kim TM, et al. Expression, circulation, and excretion profile of microRNA-21, -155, and -18a following acute kidney injury. Toxicol Sci. 2012;129(2): 256-267.
[29] Zhang Q, Xiao X, Li M, et al. Acarbose reduces blood glucose by activating miR-10a-5p and miR-664 in diabetic rats. PLoS One. 2013;8(11):e79697.
[30] Wang JF, Zha YF, Li HW, et al. Screening plasma miRNAs as biomarkers for renal ischemia-reperfusion injury in rats. Med Sci Monit. 2014;20:283-289.