睾丸转染miR-383对KM小鼠精子凋亡的影响
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摘要
目的探讨睾丸转染miRNA-383对健康可育雄性KM小鼠精子凋亡与繁殖力的影响。方法睾丸局部转染mimic miRNA-383和inhibitor miRNA-383分别获得高表达和抑制表达miRNA-383的个体,随机分为miRNA-383过表达组、miRNA-383抑制组、转染对照组、空白对照组四组。通过对转染效果、精子凋亡率、产子数和睾丸病理切片的检测来观察miRNA-383对小鼠精子凋亡和繁殖力的影响。同时对转染过程中雄鼠生活状态、体重、脏器指数、部分器官病理组织和血液生化指标进行监测,了解该操作对雄鼠机体的影响。结果成功获得在睾丸中高表达和抑制表达miRNA-383小鼠。miRNA-383过表达组精子凋亡率显著低于转染对照组(P<0.05),产子数有高于转染对照组的趋势,睾丸和肾上腺观察到一定异常,第8d和9d校正后平均体重比转染对照组显著下降(P<0.05);miRNA-383抑制表达组精子凋亡率显著高于转染对照(P<0.05),产子数对于转染对照组有下降趋势,睾丸、肺脏和肾脏观察到一定异常。结论健康可育雄性小鼠睾丸内miRNA-383表达量增加能显著降低精子凋亡率,反之则增加。miRNA-383可能对雄鼠繁殖力起重要的正调控作用,通过睾丸转染有望对雄鼠繁殖力进行人为干预。
Objective To study the influence of miRNA-383 transfection in testis on sperm apoptosis and fecundity of healthy and fertile male KM mice.Methods We transfected mimic miRNA-383 and inhibitor miRNA-383 in mice testis.Then,we got two groups of mice expressed miRNA-383 higher or lower than nomal.By testing the expressive quantity of miRNA-383,rate of sperm apoptosis,number of mice born and histological sections of testis,the influence of miRNA-383 on fecundity of fertile male mice was studied.Meanwhile,in order to find out some other effect except fecundity on mice with the operation,we monitored appearance and activities,weight,visceral index,histological changes of several organs,and blood biochemical index of male mice during transfection.Results We did get two groups of mice expressed miRNA-383 higher or lower than nomal.Compared with transfection control group,the sperm apoptosis rates of miRNA-383 overexpression group was significantly lower(P<0.05).The number born tended to be higher.The average weight at 8 day and 9 day was significantly lower(P<0.05).The testis and paranephros also observed some abnormal.At the same time,compared with transfection control group,the sperm apoptosis rates of miRNA-383 suppress expression group was significantly high(P<0.05).The number born tended to be lower.The lung,kidney and testis also observed some abnormal.Conclusion Overexpression miRNA-383 in testis of fertile mice could make sperm apoptosis rates significantly lower.Otherwise,suppress expression miRNA-383 could make it significantly higher.MiRNA-383 was positively correlated with male influence,and we might interpose it by transfectting in testis.
Objective To study the influence of miRNA-383 transfection in testis on sperm apoptosis and fecundity of healthy and fertile male KM mice.Methods We transfected mimic miRNA-383 and inhibitor miRNA-383 in mice testis.Then,we got two groups of mice expressed miRNA-383 higher or lower than nomal.By testing the expressive quantity of miRNA-383,rate of sperm apoptosis,number of mice born and histological sections of testis,the influence of miRNA-383 on fecundity of fertile male mice was studied.Meanwhile,in order to find out some other effect except fecundity on mice with the operation,we monitored appearance and activities,weight,visceral index,histological changes of several organs,and blood biochemical index of male mice during transfection.Results We did get two groups of mice expressed miRNA-383 higher or lower than nomal.Compared with transfection control group,the sperm apoptosis rates of miRNA-383 overexpression group was significantly lower(P<0.05).The number born tended to be higher.The average weight at 8 day and 9 day was significantly lower(P<0.05).The testis and paranephros also observed some abnormal.At the same time,compared with transfection control group,the sperm apoptosis rates of miRNA-383 suppress expression group was significantly high(P<0.05).The number born tended to be lower.The lung,kidney and testis also observed some abnormal.Conclusion Overexpression miRNA-383 in testis of fertile mice could make sperm apoptosis rates significantly lower.Otherwise,suppress expression miRNA-383 could make it significantly higher.MiRNA-383 was positively correlated with male influence,and we might interpose it by transfectting in testis.
引文
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[17]曲荣凤.腺嘌呤诱导小鼠精原干细胞移植受体模型的建立[D].西北农林科技大学,2014.
[2]Hayashi K,Lopes SMCDS,Kaneda M,et al.MicroRNA biogenesis is required for mouse primordial germ cell development and spermatogenesis[J].Plos One,2008,3(3):1738.
[3]Yang Q,Hua J,Wang L,et al.MicroRNA and piRNA profiles in normal human testis detected by next generation sequencing[J].PloS one,2013,8(6):66809.
[4]Lian J,Tian H,Liu L,et al.Downregulation of microRNA-383is associated with male infertility and promotes testicular embryonal carcinoma cell proliferation by targeting IRF1[J].Cell death&disease,2010,1(11):94.
[5]Lian J,Zhang X,Tian H,et al.Altered microRNA expression in patients with non-obstructive azoospermia[J].Reproductive Biology and Endocrinology,2009,7(1):1-10.
[6]Tian H,Cao Y X,Zhang X S,et al.The targeting and functions of miRNA-383are mediated by FMRP during spermatogenesis[J].Cell death&disease,2013,4(5):617.
[7]Azarbarzin S,Safaralizadeh R,Kazemzadeh M,et al.The Value of MiR-383,an Intronic MiRNA,as a Diagnostic and Prognostic Biomarker in Intestinal-Type Gastric Cancer[J].Biochemical Genetics,2017:1-9.
[8]田卉.miRNA和遗传因素在男性不育发生中的功能研究[D].中国科学技术大学,2014.
[9]Huang H,Tian H,Duan Z,et al.microRNA-383impairs phosphorylation of H2AX by targeting PNUTS and inducing cell cycle arrest in testicular embryonal carcinoma cells[J].Cellular signalling,2014,26(5):903-911.
[10]Qin Q,Shi Y,Zhao Q,et al.Effects of CD25siRNA gene transfer on high-risk rat corneal graft rejection[J].Graefe's Archive for Clinical and Experimental Ophthalmology,2015,253(10):1765-1776.
[11]Jiang Z,Chen C H,Chen Y Y,et al.Autophagic effect of programmed cell death 5(PDCD5)after focal cerebral ischemic reperfusion injury in rats[J].Neuroscience letters,2014,566:298-303.
[12]Tan C C,Zhang J G,Tan M S,et al.NLRP1inflammasome is activated in patients with medial temporal lobe epilepsy and contributes to neuronal pyroptosis in amygdala kindling-induced rat model[J].Journal of neuroinflammation,2015,12(1):18.
[13]Jiang Y,Sang Y,Qiu Q.microRNA-383mediates high glucoseinduced oxidative stress and apoptosis in retinal pigment epithelial cells by repressing peroxiredoxin 3[J].American Journal of Translational Research,2017,9(5):2374.
[14]Ma H,Liu B,Wang S,et al.MicroRNA-383is a tumor suppressor in human lung cancer by targeting endothelial PAS domain-containing protein 1[J].Cell Biochemistry and Function,2016,34(8):613-619.
[15]Fateh A,Hosseinpour Feizi M A,Safaralizadeh R,et al.Prognostic and Predictive Roles of MIR-383in Colorectal Cancer[J].2016,2016:1-14.
[16]刘风华,杨冬梓,王沂峰,等.睾丸性不育动物模型的建立[J].中华男科学杂志,2007,13(2):125-129.
[17]曲荣凤.腺嘌呤诱导小鼠精原干细胞移植受体模型的建立[D].西北农林科技大学,2014.