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第44卷第11期          潘锦堃,李超普,张 许,等. miR⁃199a⁃5p在肝脏组织中的表达及其作用的初步研究[J].
                 2024年11月                    南京医科大学学报(自然科学版),2024,44(11):1510-1516                      ·1515 ·


                A                                     B                                C
                        NC⁃mimic                               NC⁃mimic miR⁃199a⁃5p⁃mimic       NC⁃mimic
                        miR⁃199a⁃5p⁃mimic                 CD36                 100 kDa     1.5  miR⁃199a⁃5p⁃mimic
                    1.5
                   Relative mRNA level  1.0              Tubulin               180 kDa    Protein expression/Calnexin  1.0
                                                                                                 *
                                                           FAS
                                                                               45 kDa
                                                          ATGL
                    0.5
                                                                                           0.5
                                                                               60 kDa
                     0                                                                      0
                   SREBP1⁃C  ACC  FAS  CD36  ATGL  HSL CPTA⁃α                                  CD36   FAS  ATGL

                   A:The expression levels of lipid metabolism⁃related genes in Hepa1⁃6 cells after overexpression of miR⁃199a⁃5p were detected by RT⁃qPCR. B:
                The protein levels of CD36,FAS and ATGL in Hepa1⁃6 cells after overexpression of miR⁃199a⁃5p were detected by Western blot. C:Grayscale analysis
                                 *
                of protein immunoblotting. P < 0.05(n=3).
                                          图4 miR⁃199a⁃5p降低Hepa1⁃6细胞中CD36的蛋白表达
                                    Figure 4 miR⁃199a⁃5p decreased protein levels of CD36 in Hepa1⁃6 cells


                要作用   [33] 。                                           ing the global public health agenda for NAFLD:a consen⁃
                    本研究发现,在 ob/ob 小鼠肝脏 miRNA 数据库                       sus statement[J]. Nat Rev Gastroenterol Hepatol,2022,
                GEO13840中,miR⁃199a⁃5p在ob/ob小鼠肝脏中表达                     19(1):60-78
                水平增加,提示该miRNA可能在肝脏能量代谢中发                         [5] ZHOU J H,ZHOU F,WANG W X,et al. Epidemiological
                                                                       features of NAFLD from 1999 to 2018 in China[J]. Hepa⁃
                挥重要作用。此外,在高脂饮食和饥饿状态下,小
                                                                       tology,2020,71(5):1851-1864
                鼠肝脏中 miR⁃199a⁃5p 的表达水平显著增加,提示
                                                                 [6] WATT M J,MIOTTO P M,DE NARDO W,et al. The liv⁃
                在肝脏脂质积累的过程中miR⁃199a⁃5p表达水平增
                                                                       er as an endocrine organ⁃linking NAFLD and insulin re⁃
                加。进一步研究表明,过表达 miR⁃199a⁃5p 能够降                          sistance[J]. Endocr Rev,2019,40(5):1367-1393
                低肝细胞内 TAG 含量,而抑制 miR⁃199a⁃5p 会增加                 [7] GRABNER G F,XIE H,SCHWEIGER M,et al. Lipoly⁃
                肝细胞内 TAG 含量,提示 miR⁃199a⁃5p 可能参与调                       sis:cellular mechanisms for lipid mobilization from fat
                控肝细胞内脂质水平。miRDB 的预测和双荧光素                               stores[J]. Nat Metab,2021,3(11):1445-1465
                酶报告基因的实验结果显示,miR⁃199a⁃5p 靶向                      [8] LA SALA L,CARLINI V,CONTE C,et al. Metabolic dis⁃
                CD36 mRNA 的 3′ UTR 区域抑制其翻译,降低其蛋                        orders affecting the liver and heart:therapeutic efficacy of
                                                                       miRNA⁃based therapies?[J]. Pharmacol Res,2024,201:
                白水平。共转染过表达miR⁃199a⁃5p和CD36后,肝
                                                                       107083
                细胞内 TAG 含量差异无统计学意义,说明过表达
                                                                 [9] SHANG R,LEE S,SENAVIRATHNE G,et al. microR⁃
                CD36 可以抵消 miR⁃199a⁃5p 引起的细胞内 TAG 含
                                                                       NAs in action:biogenesis,function and regulation[J].
                量降低。这证实了 miR⁃199a⁃5p 通过靶向 CD36 来
                                                                       Nat Rev Genet,2023,24(12):816-833
                调节肝细胞对脂肪酸的摄取。                                    [10]DELUCAS M,SANCHEZ J,PALOU A,et al. The impact
                    本研究结果表明,在肝脏脂质积累过程中,miR⁃                            of diet on miRNA regulation and its implications for health:
                199a⁃5p的表达水平增加。miR⁃199a⁃5p表达的增加                        a systematic review[J]. Nutrients,2024,16(6):770
                抑制了 CD36 蛋白的表达,降低了肝细胞内 TAG 含                     [11]GUTBROD M J,MARTIENSSEN R A. Conserved chromo⁃
                量,有助于改善肝细胞内的脂质稳态。                                      somal functions of RNA interference[J]. Nat Rev Genet,
                                                                       2020,21(5):311-331
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