Article(id=1307013785093435898, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307013423896753139, articleNumber=null, orderNo=null, doi=10.11843/j.issn.0366-6964.2026.03.017, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1750953600000, receivedDateStr=2025-06-27, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1789545969507, onlineDateStr=2026-09-16, pubDate=1774195200000, pubDateStr=2026-03-23, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1789545969507, onlineIssueDateStr=2026-09-16, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1789545969507, creator=13701087609, updateTime=1789545969507, updator=13701087609, issue=Issue{id=1307013423896753139, tenantId=1146029695717560320, journalId=1306274286668972124, year='2026', volume='57', issue='3', pageStart='1177', pageEnd='1776', issueExtLink='null', onlineDate='null', pubDate='1774195200000', pubDateStr='2026-03-23', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1789545883392, creator='13701087609', updateTime=1789609930841, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1307282058464743901, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307013423896753139, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1307282058464743902, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307013423896753139, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1362, endPage=1372, ext={EN=ArticleExt(id=1307013785307345403, articleId=1307013785093435898, tenantId=1146029695717560320, journalId=1306274286668972124, language=EN, title=6-Phosphofructo-2-kinase/Fructose-2,6-bisphosphatase 3 (
PFKFB3) Promotes Adipogenic Differentiation of Porcine Intramuscular Preadipocytes and Screening of Its Interacting Proteins, columnId=1307007413870158054, journalTitle=Acta Veterinaria et Zootechnica Sinica, columnName=ANIMAL GENETICS AND BREEDING, runingTitle=null, highlight=null, articleAbstract=
This study aimed to investigate the regulatory role of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) in the proliferation and adipogenic differentiation of porcine intramuscular preadipocytes, screen its interacting proteins and provide clues for understanding their regulatory mechanisms. PFKFB3 was overexpressed or interfered in porcine intramuscular preadipocytes. qRT-PCR was used to detect changes in the expression of proliferation marker genes. CCK-8 and EdU assays were employed to measure changes in the number of viable cells and cells in the proliferation phase, respectively. After inducing adipogenic differentiation of porcine intramuscular preadipocytes, qRT-PCR and Oil Red O staining were used to assess its regulatory effect on adipogenic differentiation. Co-immunoprecipitation combined with liquid chromatography-tandem mass spectrometry was utilized to identify and screen proteins interacting with PFKFB3. The results showed that overexpression of PFKFB3 significantly increased the number of EdU-positive cells (P<0.01) and cell viability (P<0.01), and significantly upregulated the expression of proliferation-related genes CDK1, CCND1, PCNA, and KI67 (P<0.05). Conversely, interfering with PFKFB3 significantly inhibited cell proliferation (P<0.05). Regarding adipogenic differentiation, overexpression of PFKFB3 significantly increased the number of lipid droplets (P<0.01) and triglyceride content (P<0.01), and significantly upregulated the expression of key adipogenic genes PPARγ, CEBPα, CEBPβ, and LPL (P<0.05). Interfering with PFKFB3 significantly inhibited adipogenic differentiation (P<0.05). Furthermore, Flag-tag immunoprecipitation combined with mass spectrometry analysis identified 17 specific interacting proteins of PFKFB3. GO analysis indicated that these proteins were primarily involved in processes such as ATP binding and RNA binding. STRING protein-protein interaction network prediction revealed interactions between PFKFB3 and proteins including CHORDC1, ALAS1, ASCC3, and KCNK2. In summary, PFKFB3 positively regulates the proliferation and adipogenic differentiation of porcine intramuscular preadipocytes. Potential interacting proteins of PFKFB3 were preliminarily screened, providing new clues for in-depth analysis of the molecular mechanisms underlying porcine intramuscular fat deposition and improvement of pork quality.
, authors=Meng LI, Jiayi CHEN, Mingxin WEN, Yuqi HAN, Shaojie QIAN, Zumei MING, Xiaohong GUO, Bugao LI, authorsList=Meng LI, Jiayi CHEN, Mingxin WEN, Yuqi HAN, Shaojie QIAN, Zumei MING, Xiaohong GUO, Bugao LI, authorCompany=null, correspAuthors=Bugao LI, authorNote=null, correspAuthorsNote=
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PFKFB3)促进猪肌内前体脂肪细胞成脂分化及其互作蛋白的筛选, columnId=1307007414092456168, journalTitle=畜牧兽医学报, columnName=遗传育种, runingTitle=null, highlight=null, articleAbstract=
旨在探究6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3(6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3,PFKFB3)对猪肌内前体脂肪细胞增殖和成脂分化的调控作用,并筛选其互作蛋白,为解析其调控机制提供线索。本研究通过在猪肌内前体脂肪细胞中过表达/干扰PFKFB3,采用qRT-PCR检测增殖标志基因的表达变化,采用CCK-8、EdU试验检测活细胞及处于增殖期细胞数量的变化;诱导猪肌内前体脂肪细胞成脂分化后,通过qRT-PCR、油红O等技术,检测其对细胞成脂分化的调控作用;运用免疫共沉淀及液相色谱-质谱技术识别和筛选PFKFB3的互作蛋白。结果显示,过表达PFKFB3后EdU阳性细胞数量显著增加(P<0.01),细胞活力显著升高(P<0.01),且增殖相关基因CDK1、CCND1、PCNA、KI67表达显著上调(P<0.05);干扰PFKFB3则显著抑制细胞增殖(P<0.05)。成脂分化方面,过表达PFKFB3 可显著增加脂滴数量(P<0.01)和甘油三酯含量(P<0.01),并显著上调成脂关键基因PPARγ、CEBPα、CEBPβ、LPL等的表达(P<0.05);干扰PFKFB3显著抑制细胞的成脂分化(P<0.05)。此外,通过Flag标签免疫沉淀结合质谱分析共筛选到17个PFKFB3的特异性互作蛋白。GO分析表明这些蛋白主要参与ATP结合和RNA结合等过程;STRING蛋白互作网络预测显示PFKFB3与CHORDC1、ALAS1、ASCC3、KCNK2等蛋白存在相互作用。综上,PFKFB3可正向调控猪肌内前体脂肪细胞的增殖和成脂分化,并初步筛选出其潜在互作蛋白,为深入解析猪肌内脂肪沉积的分子机制及改善猪肉品质提供了新线索。
, authors=李萌, 陈佳仪, 文明昕, 韩雨琪, 钱少杰, 明祖梅, 郭晓红, 李步高, authorsList=李萌, 陈佳仪, 文明昕, 韩雨琪, 钱少杰, 明祖梅, 郭晓红, 李步高, authorCompany=null, correspAuthors=李步高, authorNote=
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李萌和陈佳仪为同等贡献作者
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陈佳仪,硕士,主要从事动物遗传育种与繁殖的研究,E-mail: 1321057625@qq.com
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陈佳仪,硕士,主要从事动物遗传育种与繁殖的研究,E-mail: 1321057625@qq.com
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Oil Red O staining of porcine intramuscular preadipocytesA. Adipogenic differentiation, day 2; B. Adipogenic differentiation, day 8
, figureFileSmall=DvZKYSz7+UVVNkHSQXxKxA==, figureFileBig=guHA13q/M/SOWwSan2B25w==, tableContent=null), ArticleFig(id=1307281541692941205, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=CN, label=图1, caption=
猪肌内前体脂肪细胞油红O染色结果A.成脂分化2 d;B.成脂分化8 d
, figureFileSmall=DvZKYSz7+UVVNkHSQXxKxA==, figureFileBig=guHA13q/M/SOWwSan2B25w==, tableContent=null), ArticleFig(id=1307281541797798806, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=EN, label=Fig.2, caption=
Effect of overexpression of PFKFB3 on proliferation of porcine intramuscular preadipocytesA. PFKFB3 overexpression efficiency; B. EdU staining assay, EdU-positive cells were red and nuclei were blue; C. Quantification of EdU staining results; D. Results of CCK-8 assay; E. Effects of overexpression of PFKFB3 on the expression of genes related to cell proliferation. *. P<0.05, **. P<0.01, the same as below
, figureFileSmall=8sPTVFIWj/mK59l6oL3byw==, figureFileBig=5ksRwyFEYHkUfDI/hSPQhQ==, tableContent=null), ArticleFig(id=1307281542045262743, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=CN, label=图2, caption=
过表达PFKFB3对猪肌内前体脂肪细胞增殖的影响A. PFKFB3过表达效率;B.EdU试验,阳性细胞呈红色,细胞核呈蓝色;C. EdU染色结果量化;D. CCK-8试验结果;E.过表达PFKFB3对细胞增殖相关基因表达的影响。*.P<0.05,**.P<0.01,下同
, figureFileSmall=8sPTVFIWj/mK59l6oL3byw==, figureFileBig=5ksRwyFEYHkUfDI/hSPQhQ==, tableContent=null), ArticleFig(id=1307281543714595736, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=EN, label=Fig.3, caption=
Effect of interfering with PFKFB3 on the cell proliferationA.PFKFB3 interference efficiency; B. EdU staining assay; C. Quantification of EdU staining results; D. Results of the CCK-8 assay; E. Effects of interfering with PFKFB3 on the expression of genes related to cell proliferation
, figureFileSmall=cunJvHl/O87/44Oh9AMXQg==, figureFileBig=xLmVBZGVzFO946vQWnsgBw==, tableContent=null), ArticleFig(id=1307281543798481817, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=CN, label=图3, caption=
干扰PFKFB3对细胞增殖的影响A. PFKFB3干扰效率;B.EdU试验;C. EdU染色结果量化;D. CCK-8试验结果;E.干扰PFKFB3对细胞增殖相关基因的表达变化
, figureFileSmall=cunJvHl/O87/44Oh9AMXQg==, figureFileBig=xLmVBZGVzFO946vQWnsgBw==, tableContent=null), ArticleFig(id=1307281544142414746, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=EN, label=Fig.4, caption=
Effect of overexpression of PFKFB3 on porcine intramuscular preadipocytes differentiationA, B. Oil red O staining test results; C. Effects of PFKFB3 overexpression on TG accumulation in porcine intramuscular preadipocytes; D. Effect of overexpression of PFKFB3 on genes expression related to lipogenic differentiation of cells
, figureFileSmall=u2xOaufBQLMC3QXjE4dl4g==, figureFileBig=NWUv9GSsQXF4iMKya3JUXQ==, tableContent=null), ArticleFig(id=1307281544222106523, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=CN, label=图4, caption=
过表达PFKFB3对猪肌内前体脂肪细胞分化的影响A、B.油红O染色试验结果;C.过表达PFKFB3对猪肌内前体脂肪细胞中TG含量的影响;D.过表达PFKFB3对成脂分化相关基因表达的影响
, figureFileSmall=u2xOaufBQLMC3QXjE4dl4g==, figureFileBig=NWUv9GSsQXF4iMKya3JUXQ==, tableContent=null), ArticleFig(id=1307281544305992604, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=EN, label=Fig.5, caption=
Effect of interfering with PFKFB3 on porcine intramuscular preadipocytes differentiationA, B. Oil red O staining test results; C. The effect of interfering with PFKFB3 on the TG content in porcine intramuscular preadipocytes; D. Effect of interfering with PFKFB3 on genes expression related to lipogenic differentiation of cells
, figureFileSmall=X+K+A93YbXGI2BlX5Ir6xg==, figureFileBig=ctirwldsaTqkQNqozGKUpQ==, tableContent=null), ArticleFig(id=1307281544389878685, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=CN, label=图5, caption=
干扰PFKFB3对猪肌内前体脂肪细胞分化的影响A、B.油红O染色试验结果;C.干扰PFKFB3对猪肌内前体脂肪细胞中TG含量的影响;D.干扰PFKFB3对细胞成脂分化相关基因表达的影响
, figureFileSmall=X+K+A93YbXGI2BlX5Ir6xg==, figureFileBig=ctirwldsaTqkQNqozGKUpQ==, tableContent=null), ArticleFig(id=1307281544561845150, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=EN, label=Fig.6, caption=
Results of silver staining and mass spectrometry identification analysis of PFKFB3 Pull-down proteinsA. Silver staining results of PFKFB3 interacting proteins; B. Venn diagram of PFKFB3 interacting proteins; C. GO functional annotation analysis results of PFKFB3 interacting proteins; D. Proteins interaction network diagram based on PFKFB3 interacting proteins
, figureFileSmall=m6oUYLVUCMgMezYDPaAPWg==, figureFileBig=I0Xn2LEoEM4l1Vix06w96g==, tableContent=null), ArticleFig(id=1307281544637342623, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=CN, label=图6, caption=
PFKFB3 Pull-down蛋白的银染及质谱鉴定分析结果A. PFKFB3互作蛋白银染结果;B. PFKFB3互作蛋白韦恩图;C. PFKFB3互作蛋白GO功能注释分析结果;D.基于PFKFB3互作蛋白绘制的蛋白相互作用网络关系图
, figureFileSmall=m6oUYLVUCMgMezYDPaAPWg==, figureFileBig=I0Xn2LEoEM4l1Vix06w96g==, tableContent=null), ArticleFig(id=1307281544708645792, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=EN, label=Table 1, caption=
Primer sequences of qRT-PCR
, figureFileSmall=null, figureFileBig=null, tableContent=
基因 Gene | 引物序列(5'→3') Primer sequence | 产物长度/bp Product length |
|---|
| PFKFB3 | F: ATGAGAGTCCGCAAGCAGTGT R: TCTCTAGTAGTGTTGGTGGCATCA | 110 |
| CDK1 | F: GCTTTTCAAAGCTGGCTCTGGG R: TGGATGTGGTAGATCCCAGCTTA | 104 |
| CCND1 | F: CTCCGTATCTTACTTCAAGTGCG R: CTTCTCGGCAGTCAAGGGAA | 250 |
| KI67 | F: ATCATTGACCGCTCCTTTAGGT R: GCTCGCCTTGATGGTTCCT | 104 |
| PCNA | F: GCAGAGCATGGACTCGTCTC R: TTGGACATGCTGGTGAGGTT | 150 |
| PPARγ | F: TGGGTGAAACTCTGGGAGATTC R: AGAGGTCCACAGAGCTGATTCC | 150 |
| CEBPα | F: CAAGAACAGCAACGAGTACCG R: GTCACTGGTCAACTCCAGCAC | 124 |
| CEBPβ | F: AAGAGTAAGACCAAGAAGACC R: GCTCCAGGACCTTATGCT | 139 |
| FABP4 | F: ATGTCTAGCAACAGTTTCGCC R: TCGCTTCATAGCATCTAGCTTG | 195 |
| LPL | F: CGACTCTCTGTTGAATGAAG R: TTGGCTCTGACCTTATTGATC | 141 |
| Perilipin-1 | F: TCCAGCCAAGGAAGAGTCAG R: GGAAGGTGTGTTGAGAGATGG | 125 |
| AdipoQ | F: AGGCTGGATGGCGGGTAT R: ACACTGAATGCTGAACGGTAG | 240 |
| 18S rRNA | F: ATAAACGATGCCGACTGGCGAT R: CAATCTGTCAATCCTGTCCGTGT | 245 |
), ArticleFig(id=1307281544771560353, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013785093435898, language=CN, label=表1, caption=
qRT-PCR引物信息
, figureFileSmall=null, figureFileBig=null, tableContent=
基因 Gene | 引物序列(5'→3') Primer sequence | 产物长度/bp Product length |
|---|
| PFKFB3 | F: ATGAGAGTCCGCAAGCAGTGT R: TCTCTAGTAGTGTTGGTGGCATCA | 110 |
| CDK1 | F: GCTTTTCAAAGCTGGCTCTGGG R: TGGATGTGGTAGATCCCAGCTTA | 104 |
| CCND1 | F: CTCCGTATCTTACTTCAAGTGCG R: CTTCTCGGCAGTCAAGGGAA | 250 |
| KI67 | F: ATCATTGACCGCTCCTTTAGGT R: GCTCGCCTTGATGGTTCCT | 104 |
| PCNA | F: GCAGAGCATGGACTCGTCTC R: TTGGACATGCTGGTGAGGTT | 150 |
| PPARγ | F: TGGGTGAAACTCTGGGAGATTC R: AGAGGTCCACAGAGCTGATTCC | 150 |
| CEBPα | F: CAAGAACAGCAACGAGTACCG R: GTCACTGGTCAACTCCAGCAC | 124 |
| CEBPβ | F: AAGAGTAAGACCAAGAAGACC R: GCTCCAGGACCTTATGCT | 139 |
| FABP4 | F: ATGTCTAGCAACAGTTTCGCC R: TCGCTTCATAGCATCTAGCTTG | 195 |
| LPL | F: CGACTCTCTGTTGAATGAAG R: TTGGCTCTGACCTTATTGATC | 141 |
| Perilipin-1 | F: TCCAGCCAAGGAAGAGTCAG R: GGAAGGTGTGTTGAGAGATGG | 125 |
| AdipoQ | F: AGGCTGGATGGCGGGTAT R: ACACTGAATGCTGAACGGTAG | 240 |
| 18S rRNA | F: ATAAACGATGCCGACTGGCGAT R: CAATCTGTCAATCCTGTCCGTGT | 245 |
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