共查询到20条相似文献，搜索用时 262 毫秒
1.
Keitaro Yamanouchi Katsuyuki Nakamura Shiho Takeuchi Tohru Hosoyama Takashi Matsuwaki Masugi Nishihara 《Animal Science Journal》2021,92(1):e13573
The degree of intramuscular adipose tissue accumulation is one of the factors affecting meat quality. Accumulation of adipocytes is also observed under the pathological condition of skeletal muscle such as muscular dystrophy and sarcopenia. The origin of adipocytes seen in skeletal muscle is mesenchymal progenitor cells that can give rise to both adipocytes and fibroblasts. In the present study, we demonstrated that siRNA-mediated suppression of MyoD expression in rat skeletal muscle progenitor cell culture, which comprises both myogenic satellite cells and mesenchymal progenitor cells, resulted in diminished myotube formation and an unexpected spontaneous appearance of white adipocytes. Suppressing myomaker expression also resulted in complete absence of myotube formation without reducing MyoD expression, but no adipogenesis was seen in this scenario, indicating that decline in MyoD expression rather than decreased myotube formation is necessary to induce adipogenesis. In addition, spontaneous adipogenesis induced by suppressing MyoD expression in culture was inhibited by the conditioned medium from control culture, indicating that anti-adipogenic factor(s) are secreted from MyoD-positive myogenic cells. These results indicate the presence of regulatory mechanism on adipogenesis by myogenic cells. 相似文献
2.
不同种类脂肪酸对延边黄牛骨骼肌卫星细胞成脂转分化的影响 总被引：1，自引：0，他引：1
试验旨在探究用同一浓度不同种类的脂肪酸单体（油酸、硬脂酸、棕榈油酸及棕榈酸）处理延边黄牛骨骼肌卫星细胞（BSC）,研究其对脂肪生成相关基因表达及脂滴形成的影响。从18月龄延边黄牛半膜肌中分离提取骨骼肌卫星细胞进行体外培养,在分化培养基中分别添加100 μmol/L油酸（OA）、硬脂酸（SA）、棕榈油酸（POA）和棕榈酸（PA）培养96 h,油红O染色观察脂滴生成情况,并利用实时荧光定量PCR法检测与脂肪生成相关基因过氧化物酶体增殖物激活受体γ（PPARγ）、固醇调节原件结合蛋白1（SREBP1）、硬脂酰辅酶A去饱和酶（SCD）、CCAAT/增强子结合蛋白α（C/EBPα）的表达。油红O染色结果显示,与对照组相比,所有的脂肪酸处理组细胞均有脂滴形成,油酸和棕榈油酸处理组相对于棕榈酸和硬脂酸处理组在肌管内形成的脂滴数量更多,且脂滴的形态较大。实时荧光定量PCR结果显示,在延边黄牛骨骼肌卫星细胞中添加油酸和棕榈油酸等不饱和脂肪酸增加了与脂肪合成相关基因PPARγ、SREBP1、C/EBPα的表达,抑制了SCD基因的表达;添加饱和脂肪酸（硬脂酸和棕榈酸）则在促进PPARγ、SREBP1、C/EBPα基因表达的同时也显著增加了SCD基因的表达（P < 0.05）。结果表明,添加脂肪酸可以诱导延边黄牛骨骼肌卫星细胞向脂肪细胞转分化。 相似文献
3.
Yamanouchi K Soeta C Suzuki S Hasegawa T Naito K Tojo H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2000,62(11):1213-1216
In isolating skeletal muscle satellite cells, sometimes a problem is encountered in removing contaminating nonmyogenic cells. In the present study, we constructed a novel vector, pSKA-EGFP, which achieves the expression of enhanced green fluorescent protein (EGFP) exclusively in myogenic cells under the control of skeletal alpha-actin promoter when transfected to primary cultured cells from skeletal muscle. Cells from rat skeletal muscle positive for EGFP after transfecting with pSKA-EGFP were all positive for desmin and none of the nonmyogenic cells expressed EGFP, indicating that the expression of EGFP is specific to myogenic cells. Among the cells positive for EGFP were proliferating cells, presumably satellite cells. In addition, EGFP positive cells derived from horse skeletal muscle after transfecting pSKA-EGFP in vitro formed multinuclear myotubes, indicating that myogenic expression of EGFP driven by skeletal alpha-actin was achieved also in the equine cells. These results indicated that pSKA-EGFP vector will be useful in identifying and following up the satellite cells in real time, and also permit us to isolate satellite cells in combination with fluorescence-activated cell sorting (FACS). 相似文献
4.
AMP-activated protein kinase and adipogenesis in sheep fetal skeletal muscle and 3T3-L1 cells 总被引：1，自引：0，他引：1
Marbling, or i.m. fat, is an important factor determining beef quality. Both adipogenesis and hypertrophy of existing adipocytes contribute to enhanced marbling. We hypothesized that the fetal stage is important for the formation of i.m. adipocytes and that AMP-activated protein kinase (AMPK) has a key role in adipogenesis during this stage. The objective of this study was to assess the role of AMPK in adipogenesis in fetal sheep muscle and 3T3-L1 cells. Nonpregnant ewes were randomly assigned to a control (Con, 100% of NRC recommendations, n = 7) or overfed (OF, 150% of NRC, n = 7) diet from 60 d before to 75 d after conception, when the ewes were killed. The fetal LM was collected at necropsy for biochemical analyses. The activity of AMPK was less in the fetal muscle of OF sheep. The expression of peroxisome proliferator-activated receptor (PPAR)gamma, a marker of adipogenesis, was greater in OF fetal muscle compared with Con fetal muscle. To further show the role of AMPK in adipogenesis, we used 3T3-L1 cells. The 3T3-L1 cells were incubated in a standard adipogenic medium for 24 h and 10 d. Activation of AMPK by 5-aminoimidazole-4-car-boxamide-1-beta-d-ribonucleoside dramatically inhibited the expression of PPARgamma and reduced the presence of adipocytes after 10 d of differentiation. Inhibition of AMPK by compound C enhanced the expression of PPARgamma. In conclusion, these data show that AMPK activity is inversely related to adipogenesis in fetal sheep muscle and 3T3-L1 cells. 相似文献
5.
研究旨在克隆徐淮山羊过氧化物酶体激活增殖受体(PPAR)基因的cDNA,并通过EGFP融合蛋白对该基因产物亚细胞水平定位。采用RT-PCR方法克隆徐淮山羊脂肪组织PPAR基因cDNA,并构建含有增强型绿色荧光蛋白(EGFP)报告基因的融合表达载体pEGFP-C1-PPAR,聚乙烯亚胺(PEI)介导基因转染NIH-3T3细胞,48h后荧光倒置显微镜下观察基因表达产物的分布,RT-PCR检测mRNA在体外水平的表达。结果表明:首次成功克隆出徐淮山羊PPAR基因的全序列cDNA,大小为1428bp,GenBank登录号为GU082382;构建了融合表达载体pEGFP-C1-PPAR;RT-PCR检测mRNA表达明显;EGFP-PPAR融合蛋白定位在NIT-3T3细胞质中。体外克隆的徐淮山羊PPAR基因cDNA,在单细胞水平上可表达于细胞质中,结果为进一步研究PPAR的生物学功能奠定基础。 相似文献
6.
Blanton JR Bidwell CA Sanders DA Sharkey CM McFarland DC Gerrard DE Grant AL 《Journal of animal science》2000,78(4):909-918
Cell-mediated gene transfer is a potential tool for studying muscle growth, but efficient genetic manipulation and implantation strategies have not been developed for pigs. The objectives of the present study were to determine methods for transient and stable incorporation of reporter genes into porcine muscle cells and to investigate their use for cell-mediated gene transfer in pigs. Porcine myoblasts and fibroblasts were isolated from muscle of 2-wk-old male pigs. Myogenic cell lines were identified using muscle-specific monoclonal antibodies, myotube fusion assays, and the presence of muscle-specific markers (MyoD and desmin). Four commercial cationic liposomes (lipofectAMINE, lipofectin, cellFECTIN, and DMRIE-C) were tested at different DNA:lipid ratios for their ability to transfect myoblasts and fibroblasts transiently with a luciferase reporter plasmid. LipofectAMINE resulted in the greatest (P < .01) transient luciferase activity for both cell types. Electroporation of cells for transient transfection resulted in less luciferase activity than cationic transfection. Stable transfections were conducted using a green fluorescence protein (GFP) reporter plasmid containing the neomycin resistance gene. LipofectAMINE transfection resulted in stable GFP expression in 1:16,000 myoblasts and 1:33,000 fibroblasts. Stable electroporation resulted in efficiencies that were significantly lower than established with cationic liposomes. Porcine cells were transduced with GFP using vesicular stomatitis virus glycoprotein G pseudotyped retrovirus and resulted in efficiencies of 1:1.2 for myoblasts and 1:1.1 for fibroblasts. These results show that cationic liposomes are superior to electroporation for transfection, but retroviral transduction produced stable reporter gene expression in > 80% of porcine muscle cells. Transduced GFP-positive cells were separated from GFP-negative cells by fluorescence-activated cell sorting and implanted into 2-wk-old male pigs. On d 4, implanted muscles were removed and subjected to immunodetection of GFP protein. Fibroblast implantation resulted in limited GFP expression within muscle, whereas myoblast implantation resulted in GFP within muscle fibers. This suggests that cell-mediated gene transfer is possible in porcine muscle and may be useful as an approach for studying muscle growth in pigs. 相似文献
7.
试验旨在探究过氧化物酶体增殖物激活受体（PPARγ）激动剂（环格列酮）对延边黄牛骨骼肌卫星细胞成脂转分化的影响。试验分为4组：对照组（CON，不添加环格列酮）、CL组（5 μmol/L环格列酮）、CM组（10 μmol/L环格列酮）和CH组（20 μmol/L环格列酮），利用不同浓度环格列酮处理骨骼肌卫星细胞96 h后，通过油红O染色法和甘油三酯的测定来验证脂滴的形成，并使用实时荧光定量PCR和Western blotting测定基因表达和蛋白质定量的变化来验证脂肪细胞的生成。甘油三酯测定和油红O染色显示，与对照组相比，添加环格列酮后，骨骼肌卫星细胞内有脂滴生成，且脂滴形成量和甘油积累量与环格列酮添加量呈正相关。实时荧光定量PCR和Western blotting结果表明，与对照组相比，环格列酮组显著增加了成脂转录因子PPARγ、CCAAT/增强子结合蛋白α（C/EBPα）和围脂滴蛋白-2（PLIN2）基因的表达，显著下调了成肌相关因子MyoD的表达（P<0.05），且所有蛋白与基因表达趋势保持一致，表明环格列酮可以促进牛骨骼肌卫星细胞向脂肪细胞分化。 相似文献
8.
为了选择适合牛原代骨骼肌卫星细胞的转染试剂与条件,本研究以带有绿色荧光蛋白(EGFP)的质粒pcDNA3.1-EGFP和pEGFP-C1作为外源基因,分别用Lipofectamine 3000、X-tremeGENE HP和FuGENE HD转染牛原代骨骼肌卫星细胞,通过荧光显微镜观察、实时荧光定量PCR及Hoechst 33342染色评价转染效率。结果表明,对于同一时期、同一转染试剂,pcDNA3.1-EGFP转染效果优于pEGFP-C1;不同转染试剂之间,Lipofectamine 3000和X-tremeGENE HP转染效果优于FuGENE HD。Lipofectamine 3000转染后分化72h肌管最明显,FuGENE HD次之,X-tremeGENE HP转染后细胞死亡较多。采用1.0μL Lipofectamine 3000、1.5μL Lipofectamine 3000和X-tremeGENE HP转染细胞后EGFP表达量极显著高于FuGENE HD (P<0.01),但这三者之间差异不显著(P>0.05)。细胞转染效率与EGFP定量结果基本一致,1.0、1.5μL Lipofectamine 3000转染效率较高,分别达26.07%和24.77%,极显著高于FuGENE HD(5.59%)和X-tremeGENE HP(10.87%)的转染效率(P<0.01),但二者之间无显著差异(P>0.05);X-tremeGENE HP的转染效率极显著高于FuGENE HD (P<0.01)。综合考虑转染试剂的用量、细胞毒性及经济效益,本试验初步认为牛原代骨骼肌卫星细胞转染宜采用Lipofectamine 3000,其与质粒DNA的比例为1∶1时转染效率最高。本试验结果可为原代细胞的转染提供重要的参考价值。 相似文献
9.
试验旨在探究褪黑素（MLT）对脂多糖（LPS）诱导的滩羊骨骼肌卫星细胞炎性反应的影响。选取滩羊妊娠30日龄胎儿为研究材料,使用Ⅳ型胶原酶分离获得滩羊骨骼肌卫星细胞并进行体外培养,添加相应的诱导试剂对滩羊骨骼肌卫星细胞进行诱导分化,利用免疫荧光检测骨骼肌卫星细胞表面标记物CD29、CD44、CD73及Vimentin的表达;在无胎牛血清的培养基中添加不同浓度（0、5、8和10 μg/mL）的LPS培养骨骼肌卫星细胞,利用实时荧光定量PCR （qRT-PCR）检测白细胞介素-6（IL-6）、IL-8和肿瘤坏死因子-α（TNF-α）等炎性相关因子mRNA水平变化,筛选最佳的LPS处理浓度;在无胎牛血清的培养基中添加不同浓度（0.1和0.5 μmol/L） MLT与最佳浓度LPS共培养骨骼肌卫星细胞,利用实时荧光定量PCR检测IL-6、IL-8和干扰素-γ（IFN-γ）等炎性相关因子mRNA水平变化。免疫荧光结果显示,滩羊骨骼肌卫星细胞表面标志物CD29、CD44、CD73及Vimentin表达呈阳性,且具有诱导成肌、成脂和成骨分化的特性。实时荧光定量PCR结果显示,与对照组相比,不同浓度的LPS处理均可显著增加细胞炎性相关因子基因的表达（P<0.05）,并且8 μg/mL LPS处理时炎性相关因子mRNA表达最强;当MLT与LPS共培养骨骼肌卫星细胞时,与LPS单独处理相比,0.5 μmol/L MLT与LPS共同处理组中IL-6 mRNA表达水平显著降低（P<0.05）。综上表明,MLT具有缓解LPS刺激的滩羊骨骼肌卫星细胞炎性反应的作用。本试验结果为进一步开展MLT缓解LPS诱导的骨骼肌卫星细胞炎性反应的调控机制研究奠定了基础。 相似文献
10.
11.
Wakana Izumi Yuko Takuma Ryo Ebihara Wataru Mizunoya Ryuichi Tatsumi Mako Nakamura 《Animal Science Journal》2018,89(8):1214-1219
Myogenesis is precisely proceeded by myogenic regulatory factors. Myogenic stem cells are activated, proliferated and fused into a multinuclear myofiber. Pax7, paired box 7, one of the earliest markers during myogenesis. It has been reported that Pax7 regulates the muscle marker genes, Myf5 and MyoD toward differentiation. The possible roles of Pax7 in myogenic cells have been well researched. However, it has not yet been clarified if Pax7 itself is able to induce myogenic fate in nonmyogenic lineage cells. In this study, we performed experiments using stably expressed Pax7 in 3T3‐L1 preadipocytes to elucidate if Pax7 inhibits adipogenesis. We found that Pax7 represses adipogenic markers and prevents differentiation. These cells showed decreased expression of PDGFRα, PPARγ and Fabp4 and inhibited forming lipid droplets. 相似文献
12.
九龙牦牛PPARγ基因的克隆及其表达谱分析 总被引：1，自引：0，他引：1
根据普通牛(Bos tarus)过氧化物酶体增殖物激活受体γ(PPARγ)基因序列设计引物,用成年九龙牦牛(Bos grunniens)脂肪组织总RNA,经RT-PCR扩增获得了PPARγ基因序列(GenBank登陆号:GU061328),其中cDNA的ORF为1 428 bp,编码475个氨基酸,与普通牛PPARγ氨基酸的同源性达99%,有2个氨基酸发生突变。利用半定量RT-PCR分析九龙牦牛PPARγ基因的mRNA表达特性。结果表明:在脂肪、背最长肌、心、肝、肾、脾和肺脏中均检测到PPARγ基因的表达,并且在脂肪组织中表达量极显著高于其他组织(P0.01),在肝脏和脾脏中亦有较高表达。PPARγ基因在背最长肌中的表达5.5岁九龙牦牛显著高于0.5、3.5岁和9岁以上,其表达与背最长肌的肌内脂肪含量未见显著相关性。 相似文献
13.
在包含山羊痘病毒(Goatpox virus,GPV)疫苗株复制非必需区TK(Thymidine kinase)基因和背靠背启动子的质粒PtkPP的两个启动子下游插入EGFP(增强绿色荧光蛋白)基因,构建了质粒PtkPegfpP和PtkPPegfp,这两个重组质粒分别瞬时转染LT细胞,验证了启动子P11和P7.5能成功表达EGFP基因后,在PtkPP启动子P11的下游插入gpt(黄嘌呤鸟嘌呤磷酸转移酶)基因和EGFP基因,构建重组质粒PtkPgpt-egfpP,该重组质粒和GPV同源重组获得重组病毒rGPV-PtkPgpt-egfpP,利用MPA(霉芬酸)阻断核酸代谢途径筛选到稳定表达EGFP基因的重组GPV。为进一步开展GPV活载体疫苗的研究提供了技术平台。 相似文献
14.
为了探究lncRNA TCONS_00791383对猪骨骼肌卫星细胞增殖和分化的影响。本研究利用qRT-PCR技术检测出生7 d内大白仔猪6种组织（心、脾、肺、肾、背肌和腿肌）以及猪骨骼肌卫星细胞增殖分化前后TCONS_00791383的表达水平；通过设计反义核苷酸（antisense oligonucleotides，ASO）片段在猪骨骼肌卫星细胞中对TCONS_00791383进行敲低，检测敲低TCONS_00791383之后增殖分化标志基因的表达量变化；通过trans （co-expression）对TCONS_00791383进行靶基因预测，使用DAVID对其进行GO富集和KEGG通路分析。结果显示，TCONS_00791383在猪心脏中表达量最高，在脾和肾组织中不表达。在骨骼肌卫星细胞从增殖到分化的过程中，TCONS_00791383的表达量逐渐上升，且在分化后30 h表达量达到最高。在使用ASO片段敲低TCONS_00791383之后，与对照组相比，在分化24 h，增殖标志基因Pax3、Pax7表达量显著或极显著降低（P<0.05，P<0.01），分化标志基因MyoG表达量极显著降低（P<0.01），在分化48 h，增殖标志基因Pax3表达量极显著降低（P<0.01），Pax7表达量显著降低（P<0.05），分化标志基因MyHC表达量显著降低（P<0.05）。预测得到的相关靶基因富集到AMPK、ATP等多个与骨骼肌卫星细胞增殖和分化过程相关的重要信号通路。本研究表明，lncRNA TCONS_00791383可能促进猪骨骼肌卫星细胞的增殖和分化。 相似文献
15.
Xiaoling Chen Lu Xiang Gang Jia Guangmang Liu Hua Zhao Zhiqing Huang 《Animal Science Journal》2019,90(2):255-263
A previous study demonstrated that leucine upregulates the slow myosin heavy chain mRNA expression in C2C12 cells. However, the role of leucine in slow‐twitch muscle fibers expression and mitochondrial function of porcine skeletal muscle satellite cells as well as its mechanism remain unclear. In this study, porcine skeletal muscle satellite cells cultured in differentiation medium were treated with 2 mM leucine for 3 days. Sirt1 inhibitor EX527, AMPK inhibitor compound C, and AMPKα1 siRNA were used to examine its underlying mechanism. Here we showed that leucine increased slow‐twitch muscle fibers and mitochondrial function‐related gene expression, as well as increased succinic dehydrogenase (SDH) and malate dehydrogenase (MDH) activities. Moreover, leucine increased the protein levels of Sirt1 and phospho‐AMPK. We also found that AMPKα1 siRNA, AMPK inhibitor compound C, or Sirt1 inhibitor EX527 attenuated the positive effect of leucine on slow‐twitch muscle fibers and mitochondrial function‐related gene expression. Finally, we showed that Sirt1 was required for leucine‐induced AMPK activation. Our results provide, for the first time, evidence that leucine induces slow‐twitch muscle fibers expression and improves mitochondrial function through Sirt1/AMPK signaling pathway in porcine skeletal muscle satellite cells. 相似文献
16.
Daichi IJIRI Akito SAEGUSA Tomoko MATSUBARA Yukio KANAI Miho HIRABAYASHI 《Animal Science Journal》2012,83(6):504-509
Chicks (Gallus gallus domesticus) show considerable growth of skeletal muscle during the neonatal period. The in vivo gene transfer method is useful for studying gene function and can be employed to elucidate the molecular mechanisms of skeletal muscle growth in chicks. We evaluated the following conditions for gene transfer to the skeletal muscle of neonatal chicks by electroporation: (i) voltage; (ii) age of the chick; (iii) plasmid DNA injected amount; and (iv) duration of gene expression. The results obtained from this study indicate that the most efficient gene transfer condition was as follows: 75 µg of plasmid DNA encoding β‐galactosidase was injected into the gastrocnemius muscle of chicks at 4 days of age electroporated at 50 V/cm. In addition, peak transferred gene expression was observed from 3 days to 5 days after electroporation. Our results provide optimal electroporation conditions for elucidating the gene function related to skeletal muscle growth and development in neonatal chicks. 相似文献
17.
18.
为探究肌肉生长抑制素（MSTN）对牛骨骼肌生长发育的作用机制,本研究以前期MSTN+/-蒙古牛与野生蒙古牛腿臀肌肌肉组织定量蛋白质组学与磷酸化蛋白质组学筛选获得的表达差异倍数较大的核心蛋白聚糖（DCN）为靶标,以实验室前期分离培养的牛骨骼肌卫星细胞及建立的体外诱导成肌分化模型为对象,通过对设计合成的3个DCN siRNA干扰效果的筛选,将干扰效果最显著的si-DCN-2（si-DCN）转染牛骨骼肌卫星细胞。采用实时荧光定量PCR和Western blotting方法检测增殖期（GM）牛骨骼肌卫星细胞中增殖标志因子Pax7和MyoD的mRNA水平及蛋白水平的表达变化,以及使用EdU染色的方法检测干扰DCN对细胞增殖的影响。对转染DCN siRNA的牛骨骼肌卫星细胞进行体外成肌诱导分化,通过显微镜观察牛骨骼肌卫星细胞分化第3天（DM3）的肌管形成状态,同时采用实时荧光定量PCR和Western blotting检测分化标志因子MyoG和MyHC的mRNA水平及蛋白水平的表达变化,并对DM3期肌管MyHC进行免疫荧光染色,以研究干扰DCN对细胞分化的影响。结果显示,干扰DCN表达后,增殖期牛骨骼肌卫星细胞中Pax7和MyoD的mRNA水平及蛋白水平都显著或极显著上调（P<0.05;P<0.01）,且EdU阳性细胞率显著增加（P<0.05）,表明干扰DCN表达显著促进了牛骨骼肌卫星细胞的增殖。干扰DCN表达后,牛骨骼肌卫星细胞分化第3天诱导形成的肌管直径呈现增大趋势,检测成肌分化标志因子MyoG在mRNA和蛋白水平的表达分别极显著和显著高于对照组（P<0.01;P<0.05）,MyHC在mRNA水平显著降低（P<0.05）,但在蛋白水平上极显著升高（P<0.01）,免疫荧光结果显示,下调DCN后肌管融合指数显著高于对照组（P<0.05）,说明干扰DCN表达能够促进牛骨骼肌卫星细胞的成肌分化过程。本研究结果表明,干扰DCN可以显著促进牛骨骼肌卫星细胞的增殖和成肌分化过程。研究结果为进一步开展MSTN对牛骨骼肌卫星细胞成肌分化的调控机制研究奠定了基础。 相似文献
19.
为了验证过氧化物酶体增殖物激活受体(PPAR)γ选择性抑制剂T0070907对鸡PPARγ基因表达的促进作用,本研究利用荧光定量PCR检测了不同浓度T0070907对鸡成纤维细胞(DF-1细胞)中PPARγ基因mRNA转录水平的影响,结果显示,随着T0070907浓度的增加,鸡PPARγ基因的表达量逐渐升高,并且在T0070907浓度为5μmol/L时极显著高于对照组(p<0.01);利用western blot检测了T0070907对PPARγ蛋白表达的影响,结果显示,5μmol/L的T0070907对DF-1细胞中PPARγ2蛋白的表达有一定的促进作用;利用双荧光素酶报告基因技术检测了不同浓度T0070907对PPRE报告基因活性的影响,结果显示,随着T0070907浓度的增加,DF-1细胞中PPRE报告基因活性逐渐增强,并且在T0070907浓度为0.5μmol/L时该报告基因活性显著高于对照组(p<0.05),当T0070907浓度为2μmol/L、5μmol/L时,与对照组之间的差异达到极显著水平(p<0.01);利用双荧光素酶报告基因技术检测了T0070907和罗格列酮(Rosi)对PPRE报告基因活性的影响,结果显示,单独添加T0070907或Rosi均可显著增强PPRE报告基因活性(p<0.05),同时,添加T0070907并不能降低Rosi对PPRE报告基因活性的促进作用。本研究结果表明,作为PPARγ抑制剂的T0070907对鸡PPARγ基因的mRNA的转录水平、蛋白表达及其蛋白活性均具有促进作用,推测与哺乳动物不同,鸡PPARγ可能具有其独特的表达方式和调控机制。本研究结果对完善PPARγ抑制剂在生理学及药理学上的研究、为PPARγ功能研究选择准确的拮抗剂、预防和治疗炎症相关疾病等方面具有重要意义。 相似文献
20.
为研究肌肉生长抑制素（myostatin,MSTN）对牛骨骼肌卫星细胞增殖与成肌分化的影响,本试验以牛骨骼肌卫星细胞体外诱导成肌分化模型为对象,以前期设计合成3个干扰RNA（si-MSTN-1、si-MSTN-2、si-MSTN-3）并对其进行干扰效果筛选为基础,将干扰效果极显著的si-MSTN-2（si-MSTN）转染牛骨骼肌卫星细胞,通过EdU染色法检测干扰MSTN对牛骨骼肌卫星细胞增殖的影响;进一步对干扰MSTN的牛骨骼肌卫星细胞进行体外成肌诱导分化,通过肌管形成状态和分化标志因子综合分析干扰MSTN对牛骨骼肌卫星细胞分化的影响：首先通过显微镜观察牛骨骼肌卫星细胞分化时期的肌管形成状态,然后利用实时荧光定量PCR和Western blotting技术检测牛骨骼肌卫星细胞分化标志因子MyoG和MyHC在mRNA和蛋白水平的表达情况。结果显示,干扰MSTN后,牛骨骼肌卫星细胞中EdU阳性细胞率极显著增加（P < 0.01）,说明下调MSTN表达极显著促进了牛骨骼肌卫星细胞的增殖;牛骨骼肌卫星细胞诱导分化后形成的肌管数量和直径均呈现增大趋势,牛骨骼肌卫星细胞成肌分化标志因子MyHC在mRNA和蛋白水平的表达均极显著高于对照组（P < 0.01）,说明下调MSTN表达能够促进牛骨骼肌卫星细胞的成肌分化过程。本研究结果表明,干扰MSTN可以显著促进牛骨骼肌卫星细胞的增殖及成肌分化过程。本试验结果为进一步开展MSTN对牛骨骼肌卫星细胞成肌分化的调控机制研究提供了参考。 相似文献