PGC-1α differentially regulates the mRNA expression profiles of genes related to myofiber type specificity in chicken

Previous studies on mammals showed that peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α)played a prominent role in regulating muscle fiber type transition and composition.However,the role of PGC-1αin chicken muscle has seldom been explored.To investigate the effect of PGC-1αon chicken skeletal muscles in this study,the PGC-1αgene was overexpressed or silenced in chicken primary myoblasts by using lentivirus,and then the effects of the PGC-1αgene overexpression and knockdown on the mRNA expression profile of genes related to myofiber type specificity were examined during fiber formation.The results showed that overexpression of PGC-1αfrom proliferation to differentiation was accompanied by the up-regulated expression of Pax7,MyoD,and CnAα,which was significantly(P<0.01)increased after one day of transfection(1 I).The enhancement of MyoG,MEF2 c,and MyHC SM expression lagged,which was improved significantly(P<0.01)after four days of transfection(1 I3 D).Overexpression of PGC-1αdecreased(P<0.01)the MyHC FWM expression after four days of transfection(1 I3 D),and it had no significant impact(P>0.05)on the expression of CnB1,NFATc3,and MyHC FRM during myofiber formation.The effective silence(P<0.01)of PGC-1αby lentivirus mediating short hairpin RNA(shRNA)was detected after four days of transfection(1 I3 D)in cultures,and the lack of its function in chicken primary myoblasts significantly(P<0.01)down-regulated the expression of Pax7,MyoD,CnAα,MyoG,MEF2 c,and MyHC SM,significantly(P<0.01)up-regulated the expression of MyHC FWM,and had no significant impact(P>0.05)on the expression of CnB1,NFATc3,and MyHC FRM.These results indicated that the role of PGC-1αin regulating the fiber type specificity of chicken skeletal muscles might be similar to that in mammals,which interplayed with key genes related to myocyte differentiation and calcineurin signaling pathway.

Cloning of the cDNA encoding adenosine 5'-monophosphate deaminase 1 and its mRNA expression in Japanese flounder Paralichthys olivaceus

AMP deaminase catalyzes the conversion of AMP into IMP and ammonia. In the present study, a full-length cDNA of AMPD1 from skeletal muscle of Japanese flounder Paralichthys olivaceus was cloned and characterized. The 2 526 bp cDNA contains a 5'-UTR of 78 bp, a 3'-UTR of 237 bp and an open reading frame (ORF) of 2 211 bp, which encodes a protein of 736 amino acids. The predicted protein contains a highly conserved AMP deaminase motif (SLSTDDP) and an ATP-binding site sequence (EPLMEEYAIAAQVFK). Phylogenetic analysis showed that the AMPD1 and AMPD3 genes originate from the same branch, but are evolutionarily distant from the AMPD2 gene. RT-PCR showed that the flounder AMPD1 gene was expressed only in skeletal muscle. QRT-PCR analysis revealed a statistically significant 2.54 fold higher level of AMPD1 mRNA in adult muscle (750±40 g) compared with juvenile muscle (7.5±2 g) (P<0.05). HPLC analysis showed that the IMP content in adult muscle (3.35±0.21 mg/g) was also statistically significantly higher than in juvenile muscle (1.08±0.04 mg/g) (P<0.05). There is a direct relationship between the AMPD1 gene expression level and IMP content in the skeletal muscle of juvenile and adult flounders. These results may provide useful information for quality improvement and molecular breeding of aquatic animals.

Expression of insulin-like growth factor-1 mRNA and protein level of corpora striata in ischemic side at the early stage of middle cerebral artery ischemia/reperfusion in rhesus monkeys

BACKGROUND： Insulin-like growth factor-I（IGF-1）, as one of the important members of growth factor family, participants in the regulation of many physiological functions and behaviors, having very strong neuroprotective effect. However, the expression of IGF-1 following cerebral ischemia/reperfusion is still disputed. OBJECTIVE： To observe the expression of IGF-1 and protein of corpora striata in ischemic side at the early stage of middle cerebral artery ischemia/reperfusion in rhesus monkey. DESIGN ： A completely randomized grouping design, controlled animal experiment SETTING ： Institute of Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University MATERIALS： ① Totally 17 rhesus monkeys , of either gender, aged 4 to 5 years, were enrolled . Seven rhesus monkeys observed with gene chip were randomly divided into 2 groups： sham operation group （n=3） and ischemia/reperfusion group 〈n=4〉. Ten rhesus monkeys observed with in situ hybridization and immunohistochemistry method were randomly divided into 2 groups： sham operation group 〈n=3 〉and ischemia/reperfusion group （n=7）. Rhesus monkeys observed under microscope were divided into 2 groups： sham operation group （n=6） and ischamia/reperfusion group （n=-11）.②Materials used in the experiment： cresyl violet （Sigma Company, America）; immunohistochemical reagent kit （ Huamei Bio-engineering Company）; In situ hybridization reagent kit （Boshide Bio-engineering Co.Ltd, Wuhan）; 12 800 dots chip （Boxing Company, Shanghai）. METHODS ： This experiment was carried out at the Institute of Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University from January 2001 to December 2003.① The onset area of middle cerebral artery was blocked for 2 hours, middle cerebral artery ischemia/reperfusion models were created.② After ischemia/reperfusion for 24 hours, cerebral tissue sections of rhesus monkeys were prepared and stained with cresyl violet. Image analysis was performed with 5001W image analysis software. Morphological change of corpora striata of operative side was observed in the rhesus monkeys between two groups. Total RNA was extracted from cerebral tissue. ③ Detection of gene chip： Cy3-duTP and Cy5-duTP were used to respectively perform reverse transcription labeling. The sample was reversely transcribed into cDNA, then hybridized with cDNA of cerebral tissue. Genes with the separate absolute value of cy3 and cy5〉800, cY3/cy5 〉 2（high expression） or 〈 0.5 （low expression） were found out. Those were genes with differential expression. ④ The expressions of IGF-1 mRNA and protein level of corpora striata in ischemic side of rhe- sus monkeys were detected between sham operation group and ischemia/reperfusion group at 9 and 24 hours after ischemia/reperfusion with in situ hybridization method and immunohistochemical method. Brown granules were IGF-1 protein positive cells. ⑤ Analysis of variance was used in the difference comparison of measurement data among groups. MAIN OUTCOME MEASURES ： ① Change of morphological structure of corpora striata at ischemic side in rhesus monkeys. ② Change of cerebral gene expression profiles at ischemia/reperfusion in rhesus monkeys between two groups.③ Expression of IGF-1 mRNA and protein level of corpora striata at ischemia/reperfu- sion in rhesus monkeys between two groups. RESULTS ： ① Pathological change ： Obvious pathological change of cerebral infarction appeared in the ischemia and reperfusion group, while there was no such pathological change in the sham operation group.② Change of gene expression profile ： There were 4480 genes with difference expression in the ischemia/reperfusion group and sham-operation group, in which, 260 genes had high expression and their absolute value was over 800, and 63 genes had low expression, cy3/cy5 of IGF-1 was 0.379, being relative low ex- pression. ③ IGF-1 mRNA and protein positive cell counts in corpora striata at cerebral ischemic side[IGF-1 mRNA： 〈9.72±1.18）,（9.11 ±0.76）,（14.77±0.60） counts/field：lGF-1 protein： （15.11 ±1.83）,（15.39±0.78）, （34.62±0.97）counts/field, P 〈 0.05-0.01]. CONCLUSION： IGF-1 mRNA and protein are lowly expressed in middle cerebral artery of rhesus monkeys at ischemia/reperfusion.

Lung injury and aquaporin-1mRNA expression during cardiopulmonary bypass

Objective To testify the lung injury induced by cardiopulmonary bypass(CPB) in canine model and observe the influence of CPB on the aquaporin 1 (AQP1) mRNA expression in canine lung. Methods 8 mongrel dogs were used to perform the cardiopulmonary bypass. The hearts arrested for 90 minutes with mild hypothermia and rebeated for 6 hours. The hemodynamics,the ratio of lung dry weight and wet weight,the plasmic

大鼠游泳运动后骨骼肌PGC-1α mRNA表达的时相性变化

采用实验法,检测一次长时游泳运动对大鼠骨骼肌PGC-1αmRNA表达水平的影响,以及运动后骨骼肌PGC-1αmRNA表达的时相性变化规律。将SD雄性大鼠随机分为对照组、运动后3 h组、运动后6 h组1、2 h组1、8 h组和24 h组、运动4 d后18 h组。采用SYBRGreenⅠ逆转录实时荧光定量PCR方法,测定各组大鼠骨骼肌PGC-1αmRNA表达水平。结果:PGC-1α基因在运动后6 h组、运动4 d后18 h组的表达水平高于对照组及其他组。结论:PGC-1αmRNA在游泳运动后表达的改变是有时相性的;游泳运动可以刺激骨骼肌线粒体生物发生相关因子的表达,从而调节线粒体的功能。

参芪复方对糖尿病大鼠血管PGC-1 mRNA的影响

目的:观察参芪复方对糖尿病GK大鼠血管内质网调节因子转录辅助活化因子mRNA(PGC^(-1) mRNA)的影响。方法:取5月龄雄性SPF级GK大鼠55只,随机分为空白组14只、模型组13只、中药组14只和西药组14只,有1只Wistar大鼠血糖>6.7 mmol·L^(-1)予以剔除,把剩余的14只纳入到正常组。正常组、空白组及模型组大鼠灌服生理盐水5 m L·(kg·d)^(-1),每天1次,连续给药35 d;西药组大鼠灌服阿托伐他汀钙混悬液1.6 mg·(kg·d)^(-1),每天1次,连续给药35 d;中药组大鼠灌服参芪复方混悬液1.44 g·(kg·d)^(-1),每天1次,连续给药35 d。取大鼠腹主动脉,PCR技术检测血管组织PGC^(-1) mRNA。结果:与各组大鼠比较,中药组大鼠血管PGC^(-1) mRNA明显升高,差异具有统计学意义(P<0.05)。结论:参芪复方可以调节糖尿病大鼠血管内质网PGC^(-1) mRNA水平。

The influence of aprotinin on the mRNA expression of P-selectin and ICAM-1 in the in situ lung tissue after ischemic cold storage and reperfusion

To explore the influence of aprotinin on the mRNA expression of P-selectin and ICAM-1 in lung tissue after ischemic reperfusion.Methods Thirty New Zealand white rabbits were randomly divided into 3 groups:control group,LPD group and aprotinin group.In situ rabbit lung preservation model was established.In control group,the left lower lung lobe was stored at 10℃ in a specially made lung preservation container for 2 hours and reperfused for another 2 hours.In LPD group and aprotinin group,the left lower lobe was perfused with LPD solution or aprotinin containing LPD solution,respectively,after left lung hilus was clamped.The other procedures were the same as those in control group.The lung tissue was harvested at different time intervals including preclamping lung hilus,2 hours after clamping and 2 hours after reperfusion.The mRNA expression of ICAM-1 and P-selectin in the lung tissue was detected with RT-PCR technique.Results The contents of mRNA of P-selectin at 2 hours after reperfusion in control group and LPD group were significant higher than pre-ischemia and 2 hours after champing the left lung hilus.There was no such significant difference in aprotinin group.The mRNA expression of P-selectin in aprotinin group at 2 hours after reperfusion was significantly lower than that in control group and LPD group.The ICAM-1 mRNA expression at 2 hours after Ischemia and 2 hours after reperfusion in control group and LPD group was significantly higher than the pre-ischemia and its was significantly higher than that in aprotinin group.Conclusion Aprotinin can inhibit the upregulation of the mRNA expression of P selectin and ICAM-1 after ischemia reperfusion in the lung tissue,so the addition of aprotinin in LPD solution may reduce the ischemia reperfusion injury in lung tissue.5 refs,1 tab.

Molecular cloning, sequence analysis, and cadmium stress-rated expression changes of BTG1 in freshwater pearl mussel(Hyriopsis schlegelii)

The B cells translocation gene 1 （BTG1） is a member of the BTG/TOB family of anti-proliferative genes, which have recently emerged as important regulators of cell growth and differentiation among vertebrates. Here, for the first time we cloned the full-length eDNA sequence of Hyriopsis schlegelii （Hs-BTG1）, an economically important freshwater shellfish and potential indicator of environmental heavy metal pollution, for the first time. Using rapid amplification of eDNA ends （RACE） together with splicing the EST sequence from a haemocyte eDNA library, we found that Hs-BTG1 contains a 525 bp open reading frame （ORF） encoding a 174 amino-acid polypeptide, a 306 bp 5＇ untranslated region （5＇ UTR）, and a 571 bp 3＇ UTR with a Poly（A） tail as well as a transcription termination signal （AATAAA）. Homologne searching against GenBank revealed that Hs-BTG1 was closest to Crassostrea gigas BTG1, sharing 50.57% of protein identities. Hs-BTG1 also shares some typical features of the BTG/TOB family, possessing two well-conserved A and B boxes. Clustering analysis of Hs-BTG1 and other known BTGs showed that Hs-BTG1 was also closely related to BTG1 of C. gigas from the invertebrate BTG1 clade. Function prediction via homology modeling showed that both Hs-BTG1 and C. gigas BTG1 share a similar three-dimensional structure with Homo sapiens BTG1. Tissue-specific expression analysis of the Hs-BTG1 via real-time PCR showed that the transcripts were constitutively expressed, with the highest levels in the hepatopancreas and gills, and the lowest in both haemocyte and muscle tissue. Expression levels of Hs-BTG1 in hepatopancreas （2.03-fold）, mantle （2.07-fold）, kidney （2.2-fold） and haemocyte （2.5-fold） were enhanced by cadmium （Cd2＋） stress, suggesting that Hs-BTG 1 may have played a significant role in H, schlegelii adaptation to adverse environmental conditions.

Molecular cloning and expression analysis of interleukin - 1β from Japanese sea perch(Lateolabrax japonicus )

The technology of homology cloning and anchored PCR was used to clone the IL-1β gene from the Japanese sea perch （Lateolabrax iaponicus）. The full-length cDNA of sea perch IL-1β was 1 310 bp, including a 5＇ untranslated regiop （UTR） of 136 bp, a 3＇ UTR ot 430 bp, and an ORF of 774 bp encoding a polypeptide of 258 amino acids with an estimated molecular mass of 29.31 kDa. The searches for nucleotides and protein sequence similarities with the BLAST analysis indicated that the deduced amino acid sequence of sea perch IL-1β was homological to the IL-1β in other fish species and even the mammalian. Conserved signature sequences of the IL-1β gene family were found in the sea perch IL-1β deduced amino acid sequence. Temporal expressions of the IL-1β gene in LPS or iridovirus challenged group and in control group were measured by the semi-quantitative RT-PCR. The mRNA transcripts of IL-1β could be detected in head-kidney, spleen, liver, gill and heart of the healthy individuals, and the expression level of IL-1β in head-kidney, spleen and gill was higher than that in liver and heart, but it was hard to be detected in the brain. After being stimulated by the LPS or iridovirus, the IL-1β expression in most of examined tissues was up-regulated, and also could be detected in the brain. These results indicated that the expression of sea perch IL-1β was constitutive and could be up-regulated by immune effector stimulation. Therefore the sea perch IL-1β could play a critical role in the host-pathogen interaction.

子宫内膜癌组织中IGF2BP1mRNA,PEG10mRNA表达及与增殖基因表达的相关性和预后研究

目的研究子宫内膜癌(endometrial carcinoma,EC)组织中胰岛素样生长因子2 mRNA结合蛋白1(insulinlike growth factor 2 mRNA binding protein 1,IGF2BP1)mRNA,父系表达遗传印记基因10(patrilineal expression of genetic imprinting gene 10,PEG10)mRNA表达及与增殖基因表达的相关性及预后。方法选取2017年1月~2019年1月湖北理工学院附属妇幼保健院诊治的100例EC患者。实时荧光定量PCR检测EC癌组织和癌旁组织中IGF2BP1 mRNA,PEG10 mRNA及增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)mRNA,细胞周期素D1(cyclin D1)mRNA,细胞周期蛋白依赖激酶4(cyclin dependent kinase 4,CDK4)mRNA表达。免疫组织化学检测IGF2BP1,PEG10蛋白表达。相关性采用Pearson相关分析。Kaplan-Meier曲线分析不同IGF2BP1,PEG10表达组EC患者的预后差异。COX回归分析EC患者的预后影响因素。结果EC癌组织中IGF2BP1 mRNA(1.84±0.33),PEG10 mRNA(2.12±0.40),PCNA mRNA(3.14±0.42),cyclinD1 mRNA(2.81±0.36),CDK4 mRNA(2.37±0.34)高于癌旁组织(0.78±0.21,0.91±0.25,0.74±0.13,0.67±0.21,0.59±0.18),差异具有统计学意义(t=25.652~54.588,均P<0.05)。癌组织中IGF2BP1(70.00%),PEG10(72.00%)蛋白阳性率高于癌旁组织(100%,9.00%),差异具有统计学意义(χ^(2)=75.000,82.363,均P<0.05)。EC中IGF2BP1 mRNA,PEG10 mRNA表达与PCNA mRNA,cyclinD1 mRNA,CDK4 mRNA表达呈正相关(r=0.562~0.625,均P<0.05)。EC中IGF2BP1 mRNA与PEG10 mRNA表达呈显著正相关(r=0.663,P<0.05)。FIGO分期Ⅲ期、并发淋巴结转移EC癌组织中IGF2BP1(86.49%,87.50%),PEG10(89.19%,90.63%)阳性率高于FIGO分期Ⅰ~Ⅱ期(60.32%,61.90%)、无淋巴结转移(61.77%,63.24%),差异具有统计学意义(χ^(2)=6.863~8.608,均P<0.05)。IGF2BP1阳性组患者三年总体生存率70.00%(49/70)低于阴性组的90.00%(27/30);PEG10阳性组患者三年总体生存率为69.44%(50/72),低于阴性组的92.86%(26/28),差异具有统计学意义(Log-rankχ^(2)=4.133,5.491,P=0.042,0.019)。FIGO分期Ⅲ期(OR=1.449,95%CI:1.148~1.830)、并发淋巴结转移(OR=1.442,95%CI:1.124~1.850),IGF2BP1阳性(OR=1.637,95%CI:1.239~2.163)及PEG10阳性(OR=1.576,95%CI:1.136~1.187)是影响EC患者生存预后的独立危险因素(均P<0.05)。结论EC中IGF2BP1,PEG10表达升高,两者与增殖基因表达呈正相关,是EC预后评估的肿瘤标志物。

Effect of hypoxia and glutamine or glucose deprivation on the expression of retinoblastoma and retinoblastoma-related genes in ERN1 knockdown glioma U87 cell line

The expression of retinoblastoma and several retinoblastoma-related genes was studied in glioma cell line U87 and its subline with knockdown of ERN1 (endoplasmic reticulum—nuclei-1), the main endoplasmic reticulum stress sensing and signaling enzyme. It was shown that a blockade of the ERN1 enzyme function increases the expression levels of retinoblastoma, retinoblastoma-like 1 and most retinoblastoma related genes: EID1, JARID1B, E2F1, E2F3, RBAP48 and CTIP, does not change RNF40 and RBAP46 and decreases KDM5A. We have also demonstrated that hypoxia reduces the expression levels of retinoblastoma, EID1, and E2F1 in ERN1-deficient glioma cells only. At the same time, the expression levels of retinoblastoma-like 1, E2F3, RBAP46, RBAP48 and CTIP decrease, while JARID1B and RBBP2 increase in both types of cells in hypoxic conditions, but the expression is much stronger in cells with suppressed function of ERN1. The expression level of JARID1B and KDM-5A mRNA is also enhanced in glutamine deprivation condition in both tested cell types, moreover, this effect is amplified by the blockade of the ERN1 enzyme function. The expression levels of retinoblastoma, EID1, RBAP48, and E2F3 are decreased in glutamine deprivation condition only in ERN1-deficient glioma cells, but RBL1, CTIP, RBAP46, and E2F1—in both tested cell types with more significant effect in ERN1-deficient cells. Glucose deprivation condition leads to a decrease of expression levels of retinoblastoma, RBL1, E2F3, RBAP46, and RBAP48 in both used cell types and of EID1 and E2F1 only in glioma cells with suppressed function of signaling enzyme ERN1. Thus, expression levels of retinoblastoma and most retinoblastoma-related genes are increased under a blockade of ERN1 enzyme function and significantly changed in hypoxia, glucose or glutamine deprivation conditions both in control U87 cells and ERN1-deficient cells, but inhibition of the unfolded protein response sensor ERN1 predominantly enhances these effects. Moreover, it is possible that the induction of the expression of retinoblastoma and most retinoblastoma-related genes after knockdown of ERN1 plays an important role in suppression of glioma proliferation.

Effect of Hypoxia on the Expression of a Subset of Proliferation Related Genes in IRE1 Knockdown U87 Glioma Cells

We have studied the expression of a subset of genes encoding important tumor growth related factors in U87 glioma cells with IRE1 (inositol requiring enzyme-1) knockdown as well as their hypoxic regulation. It was shown that the expression levels of activating transcription factor 6 (ATF6), clusterin (CLU), adhesion G protein-coupled receptor E5 (ADGRE5), transglutaminase?2, C polypeptide (TGM2), leukemia inhibitory factor (LIF), phosphoserine aminotransferase 1 (PSAT1), glyoxalase I (GLO1) and tetraspanin 13 (TSPAN13) are significantly down-regulated in glioma cells with the knockdown of IRE1 signaling enzyme. It was also shown that in glioma cells subjected to hypoxia, the expression levels of PSAT1, TSPAN13, EIF2AK3, and TGM2 genes were up-regulated, whereas the expression of ATF6 gene was down-regulated. At the same time, the expression levels of LIF, CLU, and ADGRE5 genes did not change in response to hypoxic treatment.?Furthermore, inhibition of IRE1, a key effector of an unfolded protein response pathway, modified the effect of hypoxia on the expression of most studied genes. Present study demonstrates that IRE1 knockdown down-regulated the expression of most studied genes and modified their hypoxic regulation and that these changes possibly contributed to the suppression of glioma growth in cells without IRE1 signaling enzyme function.

The expression of Cyclin A and p21^(cip1)in fibroblast of hypertrophic scar

Objective To study the relation of the mRNA and protein expression of CyclinA and p21cip1 in different stages hypertrophic scar fibroblast (FB) with its cell cycle,so as to provide theoretical evidence for intervention therapy of

缺氧环境对PC12细胞损伤及HIF-1 mRNA表达水平的影响

目的探讨缺氧环境对PC12细胞的损伤作用及其初步机制。方法建立PC12细胞缺氧模型;倒置相差显微镜观察细胞形态;MTT法检测正常及缺氧环境对PC12细胞的增殖;PC12细胞缺氧培养0,2,4,8,12,24和48 h后测定乳酸脱氢酶(lactate dehydrogenase,LDH);用Real-time RT-PCR检测HIF-1 mRNA的表达水平。结果缺氧培养24 h,48 h后PC12细胞增殖受到抑制(P<0.05,P<0.01);PC12细胞缺氧培养2、4、8 h后培养液中的LDH活性从2～8 h逐渐增高,8 h达到最高值(P<0.01),8 h后逐渐下降;PC12缺氧培养0～4 h HIF-1 mRNA的表达水平逐渐增高,4 h达到最高,4～48 h逐渐降低。结论缺氧环境可引起PC12细胞损伤,缺氧8 h损伤最严重。缺氧导致HIF-1基因表达水平提高,4 h时达到峰值。

鸭发育早期肝脏IGF-1 mRNA的表达特异性及其与体质量的相关性分析

选择生长速度不同的高邮鸭和金定鸭,对其胚胎期和出雏早期肝脏IGF-1 mRNA的表达规律及其与体质量的相关性进行了比较研究。采用实时荧光定量PCR方法研究鸭13、17、21、25、27胚龄和出雏后7日龄下肝脏胰岛素样生长因子1(IGF-1)mRNA的表达规律。试验结果证实:鸭肝脏IGF-1 mRNA在13胚龄已有表达。鸭早期发育过程中体质量和肝脏IGF-1 mRNA的表达水平均呈持续上升态势,高邮鸭13、21胚龄和7日龄肝脏IGF-1 mRNA的表达量极显著高于金定鸭(P<0.01),与日增体质量和绝对生长速率的变化情况相吻合。整个观察期内体质量变化和肝脏IGF-1 mRNA的表达量均呈现极显著的品种和时间特异性(P<0.01),并且受到品种和日龄交互作用的极显著影响(P<0.01)。从品种或性别的角度进行分析,均发现鸭发育早期肝脏IGF-1 mRNA的表达与体质量的变化趋势一致,且呈极显著正相关(P<0.01),提示肝脏IGF-1 mRNA的表达可能在鸭早期发育过程中发挥着重要作用。

中华稻蝗几丁质合成酶1基因的mRNA表达及功能

【目的】研究中华稻蝗(Oxya chinensis(Thunberg))几丁质合成酶1(CHS1)基因的表达特性及其在生长发育过程中的作用,从而为实现基于RNAi的蝗虫有效控制提供理论依据。【方法】根据已知中华稻蝗几丁质合成酶1基因(OcCHS1)保守区域部分cDNA序列(GenBank登录号:HM214491)设计特异性表达引物,运用RT-PCR和qPCR研究时空表达特性;采用RNA干扰技术研究其生物学功能。【结果】OcCHS1在中华稻蝗各龄期都有表达,其体表为高表达,其次是气管,其它组织部位表达量低。RNA干扰试验发现,4龄第4天若虫注射OcCHS1的双链RNA(dsRNA)后,与对照组相比,OcCHS1 mRNA表达量被沉默了70.8%,稻蝗出现蜕皮时间延迟、不能完成蜕皮或腹部皱缩死亡等现象,注射dsOcCHS1组死亡率为85.2%,与对照组(7.4%)相比差异显著。【结论】几丁质合成酶1对中华稻蝗的生长发育具有重要作用,其主要参与体表和气管几丁质的合成。采用RNA干扰技术使该基因沉默后可导致中华稻蝗死亡。

川芎嗪对发育期大鼠惊厥性脑损伤后海马caspase-1 mRNA表达的影响

目的:探讨发育期大鼠反复惊厥后海马caspase 1 mRNA表达及川芎嗪对其表达的影响。方法:162只20日龄健康SD大鼠随机分为对照组、惊厥组及川芎嗪组。采用三氟乙醚反复吸入(连续6次,每日1次)方法制作发育期大鼠惊厥模型。逆转录聚合酶链反应(RT PCR)检测各组动物反复惊厥后6 h及1、3、7 d时海马组织caspase 1 mRNA的表达,同时观察脑含水量的变化和光镜下海马区神经元病理改变,并对脑损伤进行病理半定量积分。结果:川芎嗪干预组各时间点caspase 1 mRNA表达较惊厥组均显著下调,脑含水量(7 d时除外)和脑损伤积分显著降低(P均<0.01),海马神经元水肿、变性、坏死均明显减轻。结论:川芎嗪能有效减轻惊厥性脑水肿和海马神经元病理损伤,其机制可能与抑制海马caspase 1 mRNA异常表达有关。

急性酒精暴露对人原代肝细胞HO-1 mRNA表达的影响

目的 : 研究急性酒精暴露对人原代肝细胞 HO- 1 m RNA表达的影响。方法 : 经体外灌流、分离培养人原代肝细胞 ,观察酒精对人原代肝细胞上清液中天冬氨酸氨基转移酶(AST)的释放及 GSH含量的变化 ,用 RT- PCR方法检测酒精对人原代肝细胞 HO- 1 m RNA的表达。结果 : 急性酒精暴露导致人原代肝细胞上清液中释放的 AST增加 ,并呈明显的剂量效应和时间效应关系 ;此外 ,在 1 0 0 mmol/L乙醇 2 4 h暴露下 ,肝细胞中的 GSH明显降低 ,HO- 1 m RNA表达开始明显增加 ,3～ 9h之间达到最高峰 ,随后开始降低。结论 : 在 2 5～ 1 0 0 mmol/L范围内 ,急性酒精暴露导致人原代肝细胞明显的氧化损伤 ,且影响 HO- 1 m RNA表达。

PGC-1β基因在鸡骨骼肌中的表达差异研究

为探讨PGC-1β基因在鸡骨骼肌中的表达情况,研究利用实时荧光定量PCR法检测了慢生型清远麻鸡和速生型隐性白羽鸡的比目鱼肌（主要含慢肌纤维）和趾长伸肌（主要含快肌纤维）PGC-1βm RNA的发育性表达变化。结果表明,清远麻鸡从0-9周龄,比目鱼肌和趾长伸肌的PGC-1βm RNA表达均呈＂W＂的趋势,比目鱼肌在0周龄时表达最高、5周龄时表达最低,趾长伸肌在9周龄时表达最高、7周龄时表达最低;隐性白羽鸡比目鱼肌呈＂先升后降再升＂的表达趋势,1周龄时出现峰值、7周龄时表达最低,趾长伸肌在3周龄时表达最高,显著高于其他各周龄（P〈0.05）,其他各周龄间差异不显著（P〉0.05）;相同周龄清远麻鸡比目鱼肌和趾长伸肌PGC-1βm RNA表达水平低于隐性白羽鸡。结果提示,鸡骨骼肌PGC-1βm RNA表达发育模式具有品种、年龄和表型特异性,并可能与肉品质存在关联。

断奶仔猪小肠钠葡萄糖转运蛋白1和葡萄糖转运蛋白2 mRNA表达变化及饲粮添加谷氨酰胺对其的影响

本试验旨在研究仔猪断奶后小肠黏膜钠葡萄糖转运蛋白1（SGLT1）和葡萄糖转运蛋白2（GLUT2）mRNA表达的发育性变化规律及谷氨酰胺是否对SGLT1和GLuT2mRNA的表达产生影响。选择21日龄断奶的杜×长×大仔猪69头，断奶当天屠宰3头猪，其余66头随机分成2组，每组3个重复，每个重复11头仔猪。对照组饲喂基础饲粮（不添加谷氨酰胺），试验组饲喂基础饲粮＋1％谷氨酰胺。断奶后第3、5、7、14天每组分别屠宰3头。取十二指肠、空肠和回肠黏膜样品，通过荧光定量RT—PCR方法测定SGLT1和GLUT2 mRNA的表达量。结果表明：1）SGLT1和GLUT2 mRNA的表达量在不同肠段之间无显著差异（P〉0．05）。2）十二指肠、空肠中SGLT1 mRNA表达量在断奶后先降低，断奶后第3天开始升高，第14天达峰值，回肠SGLT1 mRNA表达量在断奶后先升高，断奶后第7天较低，第14天为峰值；GLUT2 mRNA在十二指肠和空肠中的表达量从断奶后第3～7天呈现持续上升的趋势，第7天达峰值，但在第14天降至断奶当天水平，回肠GLUT2 mRNA表达量在断奶后逐渐升高，第5天达峰值，而后逐渐回落至断奶当天水平。3）饲粮添加1％谷氨酰胺对SGLT1和GLUT2 mRNA表达量无显著影响。结果提示：断奶仔猪十二指肠、空肠和回肠SGLT1和GLUT2 mRNA表达量无显著差异，但均随断奶后的时间而变化，谷氨酰胺对SGLT1和GLUT2在转录水平上无显著影响。