Effect of soothing liver therapy on oocyte quality and growth differentiation factor-9 in patients undergoing in vitro fertilization and embryo transfer

OBJECTIVE:To investigate the effect of Soothing liver therapy on infertile women undergoing in vitro fertilization and embryo transfer(IVF-ET)and to explore its mechanism.METHODS:Fifty-eight women with tubal infertility were randomized into two groups:30 in an experimental group treated with Xiaoyao powder(Shugan)plus gonadotropin-releasing hormone analog(GnRHa)/follicle-stimulating hormone(FSH)/human chorionic gonadotropin(hCG)and 28 in the control group who were treated with GnRHa/FSH/hCG only.The total gonadotropin(Gn)doses required,endometrial thickness,oocyte numbers,high quality embryo production rate and pregnancy rate of the two groups were compared.The concentration of growth differentiation factor-9(GDF-9)in follicular fluid was detected by western blotting and the expression of GDF-9 mRNA in granulosa cells was measured using reverse tran-scription-polymerase chain reaction amplification.RESULTS:In the experimental group,the Gn dose was significantly lower than that in the control group;the endometrial thickness,high quality embryo production and pregnancy rates were significantly higher and the expression of GDF-9 mRNA was also significantly higher than in the control group(all P<0.05).CONCLUSION:Shugan treatment can improve the pregnancy rate of women with tubal infertility;its mechanism is possibly related to the increased expression of GDF-9 in granulosa cells.

Porcine growth differentiation factor 9 gene polymorphisms and their associations with litter size

Growth differentiation factor 9 （GDF9） is expressed in oocytes and is thought to be required for ovarian folliculogenesis. Given this function, GDF9 may be considered as a candidate gene controlling pig ovulate rate. In this study, the complete coding sequence was cloned （encoding a 444 amino acid）, intron sequence and partial 5＇-UTR of pig GDF9. RT-PCR results showed that GDF9 mRNA is expressed in a wide range of tissues of the ruttish Erhualian pig. The expression levels of GDF9 mRNA in pituitary, ovary, uterus and oviduct are higher in the Erhualian pigs than those in Duroc pigs, especially in pituitary with a significant difference （P 〈 0.05）. Comparative sequencing revealed 12 polymorphisms, including 8 single nucleotide polymorphisms （SNPs） and one 314 bp indel in noncoding regions, and the other 3 SNPs in coding regions. Four polymorphisms, G359C, C1801T, T1806C and 314 bp indel, were developed as markers for further use in population variation and association studies. The G359C polymorphism segregates only in Chinese native pigs, Erhualian and Dahuabai, on the contrary, 314 bp indel segregates only in Duroc and Landrace. C1801T and T1806C sites seem to be completely linked and segregate in Erhualian, Dahuabai and Landrace. In a word, GDF9 may be not associated with pig litter size in extensive populations as per the studies of allele distributions of the four polymorphisms and pilot association in four breeds.

Growth differentiation factor-15 is a prognostic marker in patients with intermediate coronary artery disease

Background Growth differentiation factor-15(GDF-15)is involved in multiple processes that are associated with coronary artery disease(CAD).However,little is known about the association between GDF-15 and the future ischemic events in patients with intermediate CAD.This study was conducted to investigate whether plasma GDF-15 constituted risk biomarkers for future cardiovascular events in patients with intermediate CAD.Methods A prospective study was performed based on 541 patients with intermediate CAD(20%–70%).GDF-15 of each patient was determined in a blinded manner.The primary endpoint was major adverse cardiac event(MACE),which was defined as a composite of all-cause death,nonfatal myocardial infarction,revascularization and readmission due to angina pectoris.Results After a median follow-up of 64 months,504 patients(93.2%)completed the follow-up.Overall,the combined endpoint of MACE appeared in 134 patients(26.6%)in the overall population:26 patients died,11 patients suffered a nonfatal myocardial infarction,51 patients underwent revascularization,and 46 patients were readmitted for angina pectoris.The plasma levels of GDF-15(median:1172.02 vs.965.25 pg/m L,P=0.014)were higher in patients with ischemic events than those without events.After adjusting for traditional risk factors,higher GDF-15 levels were significantly associated with higher incidence of the composite endpoint of MACE(HR=1.244,95%CI:1.048–1.478,Quartile 4 vs.Quartile 1,P=0.013).Conclusions The higher level of GDF-15 was an independent predictor of long-term adverse cardiovascular events in patients with intermediate CAD.

GROWTH DIFFERENTIATION FACTOR-5 STIMULATES THE GROWTH AND ANABOLIC METABOLISM OF ARTICULAR CHONDROCYTES

Objective To observe the effect of growth differentiation factor-5 (GDF-5) on the growth and anabolic metabolism of articular chondrocytes. Methods The articular chondrocytes isolated from rats were treated with various concentrations of rmGDF-5, and the growth of chondrocytes measured by MTT assay, the cellular cartilage matrices formation detected sulfated glycosaminoglycan by Alcian blue staining and type Ⅱcollagen by RT-PCR. Results After 7 days culture, MTT assay showed that GDF-5 enhanced the growth of chondrocytes in a dose-dependent manner, RT-PCR showed that GDF-5 clearly induced the synthesis of type Ⅱ collagen because of the col2a1 mRNA band more and more strong in a dose-dependent. Chondrocytes were cultured with GDF-5 for 14 days, the intensity of Alcian blue staining was greatly enhanced, especially, at a high concentration of 1000ng/mL, and GDF-5 enhanced the accumulation of the Alcian blue-stainable material in a concentration-dependent manner and in a does-dependent manner. Conclusion GDF-5 enhanced the growth of mature articular chondrocytes, and stimulated the cellular cartilage matrices formation in mono-layer culture.

血清及卵泡中AMH、GDF9水平对不孕症患者生育能力的预测价值

目的探讨血清及卵泡中抗米勒管激素(anti-Müllerian hormone,AMH)、生长分化因子9(growth differentiation factor 9,GDF9)水平对不孕症患者生育能力的预测价值。方法选取2020年4月至2023年2月浙江中医药大学附属湖州中医院收治的200例不孕症患者为研究组,根据窦卵泡数目及卵巢最大平面直径分为生育能力良好组(n=114)和生育能力不良组(n=86);另选取同期132名健康体检女性为对照组。收集研究对象的一般资料与临床资料,比较研究组和对照组的血清AMH、GDF9水平;比较生育能力良好组和生育能力不良组患者的血清和卵泡AMH、GDF9水平;分析不孕症患者生育能力的影响因素;不孕症患者的血清AMH、GDF9水平与卵泡刺激素(follicle-stimulating hormone,FSH)、黄体生成素(luteinizing hormone,LH)、雌二醇水平的相关性采用Pearson相关性分析;受试者操作特征曲线分析血清AMH、GDF9水平对不孕症患者生育能力的预测价值。结果与对照组比较,研究组患者的血清AMH和GDF9水平显著降低(P<0.05)。与生育能力良好组比较,生育能力不良组患者的血清及卵泡中AMH、GDF9水平均显著降低(P<0.05)。与生育能力良好组比较,生育能力不良组患者的FSH水平、不孕年限>2年占比、促排卵次数>2次占比、卵巢间质动脉血流搏动指数、阻力指数均显著升高,获卵数、窦卵泡数目、收缩期最大血流速度和LH、雌二醇水平均显著降低(P<0.05)。不孕症患者的血清AMH和GDF9水平与FSH水平呈负相关(r<0,P<0.05),与LH和雌二醇水平呈正相关(r>0,P<0.05)。血清及卵泡AMH、血清及卵泡GDF9、获卵数、FSH、LH、窦卵泡数目、收缩期最大血流速度是不孕症患者生育能力的影响因素(P<0.05)。AMH和GDF9联合诊断不孕症患者生育能力的曲线下面积显著大于AMH、GDF9单独预测(P<0.05)。结论不孕症患者的血清及卵泡中AMH和GDF9水平与其生育能力有关,血清AMH和GDF9水平对其生育能力具有一定预测价值。

Identification of cytokines involved in hepatic differentiation of mBM-MSCs under liver-injury conditions

AIM: To identify the key cytokines involved in hepatic differentiation of mouse bone marrow mesenchymal stem cells (mBM-MSCs) under liver-injury conditions. METHODS: Abdominal injection of CCl4 was adopted to duplicate a mouse acute liver injury model. Global gene expression analysis was performed to evaluate the potential genes involved in hepatic commitment under liver-injury conditions. The cytokines involved in hepatic differentiation of mBM-MSCs was function-ally examined by depletion experiment using specifi c antibodies, followed by rescue experiment and direct inducing assay. The hepatic differentiation was characterized by the expression of hepatic lineage genes and proteins, as well as functional features. RESULTS: Cytokines potentially participating in hepatic fate commitment under liver-injury conditions were initially measured by microarray. Among the up-regulated genes determined, 18 cytokines known to closely relate to liver growth, repair and development, were selected for further identif ication. The f ibroblast growth factor-4 (FGF-4), hepatocyte growth factor (HGF) and oncostatin M (OSM) were fi nally found to be involved in hepatic differentiation of mBM-MSCs under liver-injury conditions. Hepatic differentiation could be dramatically decreased after removing FGF-4, HGF and OSM from the liver-injury conditioned medium, and could be rescued by supplementing these cytokines. The FGF-4, HGF and OSM play different roles in the hepatic differentiation of mBM-MSCs, in which FGF-4 and HGF are essential for the initiation of hepatic differentiation, while OSM is critical for the maturation of hepatocytes. CONCLUSION: FGF-4, HGF and OSM are the key cytokines involved in the liver-injury conditioned medium for the hepatic differentiation of mBM-MSCs.

BMP-4 induced proliferation and oriented differentiation of rat hepatic oval cells into hepatocytes

Objective:To explore the role of bone morphogenetic protein 4(BMP-4) in hepatic progenitor cells(HPCs).Methods:The effect of BMP-4 on rat hepatic oval cells was examined by using the WB-F344 rat hepatocytic epithelial stem-cell-like cell line.This hepatocytic cell line could exert various hepatocytc functions including the secretion of albumin and urea.Immunohistochemistry was used to examine the effects of BMP-4 and its antagonist,Noggin,on the proliferation and differentiation of these cells,cellular uptake and excretion of indocyanine green,the periodic acid-schiff(PAS) assay for glycogen storage and the expression of hepatic markers.Results:Our results showed for the first time that BMP-4 may acted as a potential inducer of hepatic differentiation in rat hepatic oval cells.Conclusions:This cell source offers a much-needed attractive and expandable source for future investigations of drug screening,stem cell technologies and cellular transplantation,in a society with increasing levels of liver disease and damage.

Effect of the gap junction blocker 1-heptanol on chondrogenic differentiation of mouse bone marrow mesenchymal stem cells in vitro

Objective：To investigate the effect of the gap junction blocker 1-heptanol on the in vitro chondrogenic differentiation of mouse bone marrow mesenchymal stem cells（MSCs） following induction by GDF-5. Methods：MSCs were isolated from mouse bone marrow and cultured in vitro. After 3 passages cells were induced to undergo chondrogenic differentiation with recombinant human GDF-5（100 ng/ml）, with or without 1-heptanol（2.5 la mol/L）. The effect of 1-heptanol on MSCs proliferation was investigated using the MTT assay. Type II collagen mRNA and protein were examined by RT-PCR and immunocytochemistry respectively, and the sulfate glycosaminoglycan was assessed by Alcian blue dye staining. Connexin43（Cx43） protein was examined by western blotting. Results：GDF-5 induced proliferation and chondrogenic differentiation of MSCs. While 1-heptanol treatment had no effect on this proliferation, it inhibited the expression of both type II collagen mRNA and protein. The Alcian blue staining revealed that 1-heptanol also inhibited the deposition of the typical cartilage extracellular matrix promoted by recombinant GDF-5. Western blotting demonstrated that 1-heptanol had no effect on the expression of Cx43. Conclusion：These results suggest that mouse bone marrow MSCs can be differentiated into a chondrogenic phenotype by GDF- 5 administration in vitro. While the gap junction blocker, 1-heptanol, did not reduce gap junction Cx43, these intercellular communication pathways clearly played an important functional role in GDF-5-induced cartilage differentiation.

GDF-9和BMP-15在PCOS卵泡发育及胰岛素抵抗中的研究进展

生长分化因子-9(growth differentiation factor-9,GDF-9)和骨形态生成蛋白-15(bone morphogenetic protein-15,BMP-15)是转化生长因子-β(transforming growth factor-β,TGF-β)超家族的组成部分,在卵巢中参与颗粒细胞和卵母细胞的增殖分化,影响卵母细胞的发育和质量,调节卵巢功能。GDF-9和BMP-15异常表达,影响卵泡发育和透明带结构,可能是导致多囊卵巢综合征(polycystic ovary syndrome,PCOS)患者不孕的重要环节。BMP-15激活Smad1/5/8信号通路,可能还与PCOS患者的胰岛素抵抗密切相关。但是,血清中GDF-9和BMP-15蛋白难以精确定量,血清GDF-9和BMP-15水平难以作为PCOS特异性诊断与治疗评估的标志。综述GDF-9和BMP-15在卵泡发育中的作用、PCOS患者卵巢GDF-9和BMP-15异常表达对卵泡发育及胰岛素抵抗的影响,探讨GDF-9和BMP-15在PCOS诊治中的潜在应用价值。

淫羊藿苷基于GDF-9/Smads对多囊卵巢小鼠排卵的作用

目的探讨淫羊藿苷(icariin,ICA)对多囊卵巢综合征(polycystic ovarian syndrome,PCOS)小鼠排卵障碍和卵泡发育的作用。方法取50只小鼠,40只用脱氢表雄酮法建立PCOS模型,随机分为模型组及ICA低剂量、中剂量和高剂量组,每组10只,剩余10只未建立模型的小鼠作为对照组。ICA低剂量、中剂量和高剂量组分别按25、50、75 mg·kg^(-1)腹腔注射0.5 mL ICA(溶于9 g·L^(-1)氯化钠注射液0.5 mL中),对照组和模型组腹腔注射9 g·L^(-1)氯化钠注射液,比较各组小鼠激素水平、脏器指数、卵巢形态学差异、卵母细胞凋亡率、生长分化因子9(growth differentiation factor-9,GDF-9)、骨形成蛋白Ⅱ型受体(bone morphogenetic protein typeⅡreceptor,BMPR-Ⅱ)、激活素受体样激酶5(activin receptor like kinase 5,ALK5)、磷酸化Smad蛋白2(phospho-Smad protein 2,p-Smad2)及p-Smad3蛋白水平的差异。结果HE结果显示ICA各剂量组小鼠各级卵泡数量增多,颗粒细胞层数增加、囊性卵泡数量减少;ICA各剂量组睾酮激素(testosterone,T)、促黄体生成激素(luteinizinghor mone,LH)、抗缪勒管激素(anti-Müllerian hormone,AMH)水平、卵巢指数、囊性卵泡数量以及卵泡细胞凋亡率较模型组降低,卵泡刺激素(follicle stimulating hormone,FSH)、雌二醇(estradiol,E 2)水平、子宫指数、生长卵泡数量、黄体数量、GDF-9、BMPR-Ⅱ、ALK5、p-Smad2、p-Smad3蛋白表达水平较模型组升高(P<0.05);T、LH、AMH水平及卵巢指数、囊性卵泡数量、卵泡细胞凋亡率与ICA呈剂量依赖性降低,E 2、FSH水平、子宫指数、生长卵泡数量、黄体数量以及GDF-9、BMPR-Ⅱ、ALK5、p-Smad2和p-Smad3蛋白的表达水平与ICA呈剂量依赖性升高(P<0.05)。结论ICA能有效调节PCOS小鼠生殖激素水平,改善内分泌代谢,促进卵泡发育、成熟,减少卵泡凋亡,改善排卵,可能与调控GDF-9/Smads信号通路蛋白的表达有关。

山羊生长分化因子9基因外显子2的PCR-SSCP分析

目的寻找与产羔数相关的遗传标记,为山羊高繁殖力的标记辅助选择提供科学依据。方法以控制Belclare绵羊和Cambridge绵羊高繁殖力的生长分化因子9(growthdifferentiationfactor9,GDF9)基因为候选基因,根据绵羊GDF9基因序列设计4对引物,采用PCR-SSCP技术检测GDF9基因外显子2在高繁殖力山羊品种(济宁青山羊)以及低繁殖力山羊品种(波尔山羊、文登奶山羊、辽宁绒山羊、北京本地山羊)中的单核苷酸多态性,同时研究该基因对济宁青山羊高繁殖力的影响。结果山羊与绵羊的GDF9基因外显子2核苷酸序列同源性为99%(955/965)。引物1和引物4存在多态性。对于引物1扩增片段,5个山羊品种均检测到AA、AB和BB3种基因型,测序分析发现外显子2第26bp处有1个G→A的单碱基突变,但没有引起氨基酸改变;济宁青山羊A等位基因频率为0.9128,B等位基因频率为0.0872;AA基因型济宁青山羊产羔数最小二乘均值比AB基因型的多0.54只(P<0.01),比BB基因型的多0.63只(P<0.01)。对于引物4扩增片段,5个山羊品种均检测到CC、CD和DD3种基因型,测序分析发现外显子2第792bp处有1个G→A的单碱基突变并且导致缬氨酸改变为异亮氨酸;济宁青山羊C等位基因频率为0.9266,D等位基因频率为0.0734;CC基因型济宁青山羊产羔数最小二乘均值比CD基因型的多0.57只(P<0.01),比DD基因型的多0.62只(P<0.01)。结论初步表明GDF9基因可能是控制济宁青山羊多胎性能的一个主效基因或是与之存在紧密遗传连锁的分子标记。

绵羊GDF9基因PCR-SSCP分析

生长分化因子 9(GDF9)是由卵母细胞分泌的一种生长因子 ,它对早期卵泡的生长和分化起重要的调节作用。采用PCR SSCP技术分析了GDF9基因在小尾寒羊、湖羊、多赛特羊和萨福克羊 4个绵羊品种的多态性。结果表明 :GDF9基因在两对引物扩增片段中均存在PCR SSCP多态性。对于引物 1扩增片段 ,4个绵羊品种均检测到AA基因型 ,AB基因型只出现在湖羊、多赛特羊和萨福克羊中 ,仅在萨福克羊中检测到BB基因型 ;在 4个绵羊品种中 ,A等位基因频率明显高于B等位基因频率。对于引物 2扩增片段 ,4个绵羊品种均检测到AA基因型 ,AB基因型只出现在湖羊、多赛特羊和萨福克羊中 ,4个绵羊品种均没有检测到BB基因型 ;在 4个绵羊品种中 ,AA基因型频率最高 ,A等位基因频率明显高于B等位基因频率。引物 1的多态性片段测序分析表明 :位于GDF9基因cDNA第15 2处发生了单碱基的改变 (A→G) ,并导致了氨基酸的改变 (天冬酰胺→天冬氨酸 )。

小尾寒羊高繁殖力候选基因BMP15和GDF9的研究

以控制Belclare和Cambridge绵羊高繁殖力的骨形态发生蛋白 15 (bonemorphogeneticprotein 15 ,BMP15 )基因和生长分化因子 9(growthdifferentiationfactor 9,GDF9)基因为候选基因 ,采用PCR RFLP技术检测BMP15基因和GDF9基因在高繁殖力绵羊品种 (小尾寒羊、湖羊 )以及低繁殖力绵羊品种 (多赛特羊、特克塞尔羊、德国肉用美利奴羊 )中的单核苷酸多态性 ,同时研究这两个基因对小尾寒羊高繁殖力的影响。结果表明 :在 5个绵羊品种中都没有检测到GDF9基因的G8突变 (C→T) ,也没有检测到BMP15基因的B4突变 (G→T)。高繁殖力的小尾寒羊在BMP15基因编码序列第 718位碱基处发生了与Belclare绵羊和Cambridge绵羊相同的B2突变 (C→T) ,而其余 4个绵羊品种则没有发生这种突变。对于BMP15基因的B2突变 ,在小尾寒羊中检测到AA、AB两种基因型 ,A等位基因频率为 0 734,B等位基因频率为 0 2 6 6。小尾寒羊与其余 4个绵羊品种间B2突变基因型分布差异极显著 (P <0 0 0 1)。突变杂合基因型 (AB)小尾寒羊平均产羔数比野生纯合基因型 (AA)多 0 6 2只 (P <0 0 1)。研究结果表明 ,BMP15B2突变对小尾寒羊高繁殖力影响作用十分明显 。

生长分化因子9基因的研究进展

转化生长因子β( TGF-β)超家族成员对卵巢卵泡的发育起着重要的作用 ,在这个超家族中发现 1个新成员生长分化因子 ( GDF-9) ,它对早期卵泡和卵母细胞的生长是必需的。本文主要阐述了生长分化因子 9基因结构和功能、基因克隆以及在小鼠、人类和绵羊卵巢中的作用机理 ,并讨论了该基因对繁殖性能的影响。

生长分化因子9基因及其在生殖中的作用

生长分化因子9是卵母细胞分泌的一种生长因子,它对卵泡的生长分化起着重要作用。文章介绍了生长分化因子9的结构、功能和调控,生长分化因子9基因的克隆及基因结构、发育性表达、定位和多态性,并讨论了该基因与哺乳动物繁殖性能的关系。

补肾调经方对输卵管性不孕症患者体外受精—胚胎移植超排卵周期GDF-9的影响

目的研究补肾调经方对提高体外受精—胚胎移植(in vitrofertilization-embryo transfer,IVF-ET)超排卵周期所获卵和胚胎质量的作用及其机制。方法将58例因输卵管性不孕行IVF-ET患者随机分为中药+控制性超排卵组(治疗组,30例)和单纯控制性超排卵组(对照组,28例)。应用Western blot法测定取卵日成熟卵泡卵泡液中生长分化因子-9(growth differentiation factor-9,GDF-9)的含量;用RT-PCR法检测颗粒细胞中GDF-9的表达;并比较两组的促性腺激素(gonadotropin,Gn)用药量、获卵数、受精率、卵裂率、优质胚胎率及妊娠率等。结果治疗组卵泡液中GDF-9水平及颗粒细胞中GDF-9的表达均显著高于对照组(P<0.05);治疗组的获卵数、受精率、优质胚胎率及妊娠率均显著高于对照组(P<0.05);Gn用量及卵裂率两组比较差异无统计学意义。结论补肾调经方可以提高IVF-ET妊娠率,其作用机制可能与调控卵泡液和颗粒细胞GDF-9水平有关。

检测恶性腹水中VEGF、CD44v6和MMP-2、MMP-9的临床意义

背景及目的:恶性腹水中找到癌细胞有确诊意义,但阳性率很低。因此从腹水中寻找理想的癌性标记物已成为当前研究的主要课题。本课题拟通过检测腹水中血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)、细胞粘附分子CD44v6、基质金属蛋白酶-2,-9(matrixmetalloproteinase-2,-9,MMP-2,-9)的水平并进行分析,以期为临床腹水诊治提供科学依据。方法:收集肝硬化腹水36例、结核性腹水8例、恶性腹水23例。采用ELISA法检测VEGF、CD44v6含量,用明胶酶谱法检测MMP-2,-9活性。结果:恶性腹水中VEGF、CD44v6含量分别为(640.74±264.81)ng/L、(89.22±38.20)μg/L,明显高于肝硬化腹水犤(67.05±51.91)ng/L,(44.79±18.02)μg/L犦和结核性腹水犤(88.25±24.12)ng/L,(50.25±12.57)μg/L犦(P<0.01)。肝硬化腹水、结核性腹水不能检出MMP-2,-9,而恶性腹水MMP-2检出率为87.0%,MMP-9检出率为78.3%。结论:恶性腹水中VEGF、CD44v6水平显著升高而MMP-2,-9阳性。联合检测腹水中VEGF、CD44v6、MMP-2,-9的水平可作为临床良、恶性腹水的鉴别诊断依据之一。

骨形态发生蛋白9定向诱导多潜能干细胞成骨分化

为确认和评价骨形态发生蛋白9(bone morphogenetic protein9,BMP9)定向诱导多潜能干细胞成骨分化的能力,以鼠间充质干细胞C3H10、小鼠胚胎成纤维细胞(mouse embryonic fibroblasts,MEFs)和骨髓基质细胞(bone marrow stromalcell,BMSC)三种多潜能干细胞为目标细胞,用重组腺病毒的方法将BMP9导入细胞,通过荧光素酶报告基因实验、碱性磷酸酶(alkaline phosphatase,ALP)定量测定、钙盐沉积实验、realtime PCR、动物实验和组织化学染色等方法,观察BMP9对于多潜能干细胞成骨分化的定向诱导作用.结果提示,BMP9能诱导C3H10、MEFs和BMSC细胞ALP的表达,且具有剂量依赖性.BMP9在体外能够促进C3H10细胞和MEFs细胞的钙盐沉积.经BMP9刺激后,C3H10细胞成骨相关基因ALP、Runx2、骨桥素(osteopontin,OPN)和骨钙素(osteocalcin,OC)的mRNA水平均增加.荧光素酶报告基因实验证实,BMP9可以活化Smad和成骨关键基因Runx2.动物实验和组织化学染色检查显示,BMP9可以诱导C3H10细胞在裸鼠皮下异位成骨,因此,BMP9具有定向诱导多潜能干细胞成骨分化的能力.

生长分化因子-9在大鼠卵巢中的表达

目的:探讨生长分化因子- 9(GDF - 9)在大鼠卵巢中的表达部位。方法:选择不同发育时期的大鼠卵巢,采用免疫组化和Westernblot技术检测大鼠卵泡各发育阶段GDF - 9蛋白表达情况及分布规律。结果:免疫组化方法显示,出生0～2d卵巢中未见GDF - 9表达,出生3d卵巢中见部分原始卵泡卵母细胞GDF - 9表达阳性,以后从初级卵泡到发育成熟的卵泡卵母细胞均明显可见GDF - 9蛋白表达。Westernblot技术进一步提示,出生0～2d卵巢中无GDF - 9蛋白形成,第3d开始有GDF - 9蛋白产生,以后在卵巢发育的各时期均有GDF - 9蛋白分泌。结论:GDF - 9在卵巢发育的早期即开始产生,以后在各阶段卵巢中持续表达,提示卵母细胞通过产生GDF -

绵羊GDF9基因FecTT突变检测

FecTT是冰岛Thoka绵羊GDF9基因外显子2编码区1279位发生的A→C突变(A1279C),导致编码GDF9蛋白C末端成熟肽第109位丝氨酸改变为精氨酸(S109R),该突变纯合子母羊不育,杂合子母羊产羔数比野生型多0.6只。本研究采用PCR-RFLP方法分析了GDF9基因FecTT突变在小尾寒羊、洼地绵羊、湖羊、考力代、白萨福克、黑萨福克、东弗里生、杜泊绵羊、特克塞尔、多赛特和中国美利奴11个绵羊品种中的分布。结果表明在11个绵羊品种中都没有检测到GDF9基因的FecTT突变,提示GDF9基因影响Thoka绵羊高繁殖力的FecTT突变对以上11个绵羊品种的繁殖力均没有显著影响。