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表3 靶向Snail1的小鼠血浆外泌体差异miRNA
Table 3 Sequencing results of different plasma exosomal miRNA targeting Snail1

差异 正反 长
miRNA 来源 染色体 起始 终止 |log3FC| 成熟体序列（5′→3′）
表达 链 度
mmu⁃let⁃7a⁃1⁃3p TargetScan up chr13 48538188 48538209 - 21 0.312 CUAUACAAUCUACUGUCUUUCC
mmu⁃let⁃7b⁃3p TargetScan up chr15 85707379 85707400 + 21 0.322 CUAUACAACCUACUGCCUUCCC
mmu⁃let⁃7c⁃2⁃3p TargetScan up chr15 85706664 85706685 + 21 0.312 CUAUACAAUCUACUGUCUUUCC
mmu⁃let⁃7g⁃3p TargetScan up chr9 106178902 106178923 + 21 1.907 ACUGUACAGGCCACUGCCUUGC
mmu⁃miR⁃296⁃5p TargetScan up chr2 174267093 174267113 - 20 1.0875 AGGGCCCCCCCUCAAUCCUGU
mmu⁃miR⁃3094⁃3p TargetScan up chr4 40993738 40993758 + 20 3.764 CCUUUAAAUUGUGUCCUCAAG
mmu⁃miR⁃34a⁃5p TargetScan up chr4 150068473 150068494 + 21 0.322 UGGCAGUGUCUUAGCUGGUUGU
mmu⁃miR⁃34c⁃5p TargetScan up chr9 51103076 51103098 - 22 0.737 AGGCAGUGUAGUUAGCUGAUUGC
mmu⁃miR⁃493⁃5p TargetScan up chr12 109580243 109580261 + 18 1.585 UUGUACAUGGUAGGCUUUC
mmu⁃miR⁃98⁃3p TargetScan up chrX 151913287 151913308 + 21 1.000 CUAUACAACUUACUACUUUCCU
mmu⁃miR⁃221⁃5p TargetScan down chrX 19146344 19146369 - 25 -0.539 ACCUGGCAUACAAUGUAGAUUUCUGU
mmu⁃miR⁃3061⁃3p TargetScan down chr11 52126804 52126825 + 21 -2.121 CUACCUUUGAUAGUCCACUGCC
mmu⁃miR⁃30b⁃5p TargetScan down chr15 68337469 68337490 - 21 -0.366 UGUAAACAUCCUACACUCAGCU
mmu⁃miR⁃30c⁃5p TargetScan down chr1； 23291714； 23291736； +； 22； -0.466 UGUAAACAUCCUACACUCUCAGC
0 0 0 chr4 120769584 120769606 - 22 0 0
mmu⁃miR⁃30e⁃5p TargetScan down chr4 120772660 120772681 - 21 -0.500 UGUAAACAUCCUUGACUGGAAG
mmu⁃miR⁃449a⁃5p TargetScan down chr13 113037549 113037570 + 21 -2.706 UGGCAGUGUAUUGUUAGCUGGU
mmu⁃miR⁃615⁃5p TargetScan down chr15 103014926 103014947 + 21 -5.291 GGGGGUCCCCGGUGCUCGGAUC
mmu⁃let⁃⁃7e⁃5p MiRanda up chr17 17830366 17830387 + 21 0.494 UGAGGUAGGAGGUUGUAUAGUU
mmu⁃let⁃7f⁃5p MiRanda up chr13； 48537889； 48537910； -； 21； 0.405 UGAGGUAGUAGAUUGUAUAGUU
0 0 chrX 151912353 151912374 + 21 0 0
mmu⁃let⁃7k MiRanda up chr5 147283257 147283275 - 18 0.935 UGAGGUAGGAGGUUGUGUG
mmu⁃miR⁃1961 MiRanda up chr5 92788465 92788481 - 16 1.100 UGAGGUAGUAGUUAGAA
mmu⁃miR⁃7030⁃5p MiRanda up chr5 115645578 115645599 + 21 3.764 UGGGGAGGUGGCUGUGCUAGUC
mmu⁃miR⁃7074⁃5p MiRanda up chr8 105951672 105951694 - 22 7.465 UGGGAAGAGGCUAGGGCUCCAGU
mmu⁃miR⁃872⁃3p MiRanda up chr4 94665206 94665227 + 21 0.322 UGAACUAUUGCAGUAGCCUCCU
mmu⁃miR⁃200c⁃3p MiRanda down chr6 124718324 124718346 - 22 -0.282 UAAUACUGCCGGGUAAUGAUGGA
mmu⁃miR⁃330⁃5p MiRanda down chr7 19181486 19181507 + 21 -0.537 UCUCUGGGCCUGUGUCUUAGGC
mmu⁃miR⁃370⁃3p MiRanda down chr12 109618305 109618326 + 21 -1.346 GCCUGCUGGGGUGGAACCUGGU
mmu⁃miR⁃5124a MiRanda down chr13 40865801 40865820 + 19 -0.678 GGUCCAGUGACUAAGAGCAU
mmu⁃miR⁃6935⁃5p MiRanda down chr11 120347387 120347408 - 21 -1.121 UGUGGGAGGCUCAGGGCCCCUG
mmu⁃miR⁃6980⁃5p MiRanda down chr18 37990919 37990940 - 21 -1.121 GUGGGGGGGGAGGCUAGGUUAG
mmu⁃miR⁃7093⁃3p MiRanda down chrX 134757711 134757731 + 20 -2.706 UUUCCAUCUGUCAUCCUGCAG
mmu⁃miR⁃7669⁃3p MiRanda down chr3 90011153 90011175 - 22 -3.121 GAGCUGGGUGUGGUGGCACAUGC
相比，db/db 组小鼠肾组织 mmu⁃miR⁃296⁃5p 降低， Snail1的靶向关系
mmu⁃miR⁃34a⁃5p 升高（P=0.001，P=0.011），差异有 TargetScan 软件预测 mmu⁃miR⁃296⁃5p 与 Snail1
统计学意义（图 2）。进一步利用小鼠血浆外泌体 3′UTR 的结合位点位于 Snail1 3′UTR 区 596~603 位
行 RT⁃qPCR 验证，发现 db/db 组小鼠血浆外泌体中 置（图4）。双荧光素酶报告基因检测实验结果显示：
mmu⁃miR⁃296⁃5p 表达下降，mmu⁃miR⁃34a⁃5p 表达 与miR⁃296⁃5p mimics NC组相比，miR⁃296⁃5p mimics
升高（P=0.011，P=0.013），差异有统计学意义（图 组 Snail1⁃WT 细胞的荧光素酶活性强度明显降低
3）。结合|log2FC|值，选取miR⁃296⁃5p为本研究目标 （P < 0.001）；Snail1⁃MUT 细胞的荧光素酶活性强
miRNA。 度无明显变化（P=0.060）；Snail1⁃WT 细胞 miR⁃296⁃
2.4 双荧光素酶实验检测mmu⁃miR⁃296⁃5p与Mus⁃ 5p mimics 组与 Snail1⁃MUT 细胞 miR⁃296⁃5p mimics