目的:探究芪丹通脉片(QDTM)对阿霉素诱导扩张型心肌病(DCM)大鼠的治疗作用及其对长链非编码RNA(lncRNA)X-无活性特异性转录物(XIST)表达的影响。方法:将大鼠分为Con组(n=12)、DCM组(n=13)、L-QDTM组(n=13)、M-QDTM组(n=13)、H-QDTM组(n=...目的:探究芪丹通脉片(QDTM)对阿霉素诱导扩张型心肌病(DCM)大鼠的治疗作用及其对长链非编码RNA(lncRNA)X-无活性特异性转录物(XIST)表达的影响。方法:将大鼠分为Con组(n=12)、DCM组(n=13)、L-QDTM组(n=13)、M-QDTM组(n=13)、H-QDTM组(n=13)。Con组大鼠为正常对照大鼠,其他组大鼠均为阿霉素诱导的扩张型心肌病模型大鼠。Con组和DCM组大鼠灌胃生理盐水,L-QDTM组、M-QDTM组和H-QDTM组大鼠分别灌胃500、1000和2000 mg/kg/d的芪丹通脉片浸膏干粉,每日给药1次,共4周。治疗结束后,分别检测各组大鼠的心功能参数,血清心肌损伤指标和心肌组织氧化应激指标。通过心肌组织苏木素伊红(HE)染色、Masson三色染色和TUNEL染色观察心肌形态变化、纤维化和细胞凋亡情况。通过RT-qPCR检测心肌组织中XIST、collagen I、collagenⅢ、TGF-β1、Bax和Bcl-2的转录水平。结果:与Con组比较,DCM组大鼠的左室射血分数(LVEF)和左室短轴缩短率(FS)降低,左室舒张末期内径(LVIDd)和左心室收缩末期内径(LVIDs)升高,乳酸脱氢酶(LDH)、肌酸激酶(CK)和心肌肌钙蛋白I(cTnI)升高;心肌出现明显损伤,纤维化面积升高;collagen I、collagenⅢ和TGF-β1的m RNA水平均升高,TUNEL阳性率升高;Bax m RNA水平升高,Bcl-2 m RNA水平降低,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)水平降低,丙二醛(MDA)水平升高,XIST水平升高(均P<0.05)。与DCM组比较,L-QDTM组、M-QDTM组和H-QDTM组的LVEF和FS均升高,LVIDd和LVIDs均降低,LDH、CK和cTnI均降低;心肌损伤减轻,纤维化面积降低;collagen I、collagenⅢ和TGF-β1的m RNA水平均降低,TUNEL阳性率降低;Bax m RNA水平降低,Bcl-2 m RNA水平升高,SOD和CAT水平升高,MDA水平均降低,XIST水平降低(均P<0.05)。结论:本研究表明芪丹通脉片在治疗阿霉素诱导扩张型心肌病大鼠中效果显著,其机制可能与抑制lncRNA XIST有关。展开更多
In somatic cell nuclear transfer (SCNT) technologies,the donor cell’s nuclei need to be epigenetically reprogrammed for embryonic development. The incomplete reprogramming of donor cell nuclei has been implicated as ...In somatic cell nuclear transfer (SCNT) technologies,the donor cell’s nuclei need to be epigenetically reprogrammed for embryonic development. The incomplete reprogramming of donor cell nuclei has been implicated as a primary reason for the low efficiency of SCNT. DNA methylation is a major epige-netic modification of the genome that regulates crucial aspects of genome function,including estab-lishment of genomic imprinting. In order to make sure whether the DNA methylation reprogramming is efficient in SCNT animals,we analyzed the DNA methylation status of two imprinting genes,H19 and Xist,in lungs of deceased SCNT bovines that died within 48 h of birth using bisulfite sequencing analysis. Our findings demonstrated that cloned bovines showed significantly lower DNA methylation of H19 than controls (P<0.05),and three tested CpGs sites (1,2,3) exhibited unmethylation in one cloned bovine (9C3); however,Xist showed similar DNA methylation levels between clones and con-trols,and both showed hypermethylation (96.11% and 86.67%).展开更多
The limited knowledge of genomic noncoding and regulatory regions has restricted our ability to decipher the genetic mechanisms underlying complex traits in pigs. In this study, we characterized the spatiotemporal lan...The limited knowledge of genomic noncoding and regulatory regions has restricted our ability to decipher the genetic mechanisms underlying complex traits in pigs. In this study, we characterized the spatiotemporal landscape of putative enhancers and promoters and their target genes by combining H3K27ac-targeted Ch IP-Seq and RNA-Seq in fetal(prenatal days 74–75) and adult(postnatal days 132–150) tissues(brain, liver, heart, muscle and small intestine) sampled from Asian aboriginal Bama Xiang and European highly selected Large White pigs of both sexes. We identified 101,290 H3K27ac peaks, marking 18,521promoters and 82,769 enhancers, including peaks that were active across all tissues and developmental stages(which could indicate safe harbor locus for exogenous gene insertion) and tissue-and developmental stage-specific peaks(which regulate gene pathways matching tissue-and developmental stage-specific physiological functions). We found that H3K27ac and DNA methylation in the promoter region of the XIST gene may be involved in X chromosome inactivation and demonstrated the utility of the present resource for revealing the regulatory patterns of known causal genes and prioritizing candidate causal variants for complex traits in pigs. In addition, we identified an average of 1,124 super-enhancers per sample and found that they were more likely to show tissue-specific activity than ordinary peaks. We have developed a web browser to improve the accessibility of the results(http://segtp.jxau.edu.cn/pencode/?genome=sus Scr11).展开更多
文摘目的:探究芪丹通脉片(QDTM)对阿霉素诱导扩张型心肌病(DCM)大鼠的治疗作用及其对长链非编码RNA(lncRNA)X-无活性特异性转录物(XIST)表达的影响。方法:将大鼠分为Con组(n=12)、DCM组(n=13)、L-QDTM组(n=13)、M-QDTM组(n=13)、H-QDTM组(n=13)。Con组大鼠为正常对照大鼠,其他组大鼠均为阿霉素诱导的扩张型心肌病模型大鼠。Con组和DCM组大鼠灌胃生理盐水,L-QDTM组、M-QDTM组和H-QDTM组大鼠分别灌胃500、1000和2000 mg/kg/d的芪丹通脉片浸膏干粉,每日给药1次,共4周。治疗结束后,分别检测各组大鼠的心功能参数,血清心肌损伤指标和心肌组织氧化应激指标。通过心肌组织苏木素伊红(HE)染色、Masson三色染色和TUNEL染色观察心肌形态变化、纤维化和细胞凋亡情况。通过RT-qPCR检测心肌组织中XIST、collagen I、collagenⅢ、TGF-β1、Bax和Bcl-2的转录水平。结果:与Con组比较,DCM组大鼠的左室射血分数(LVEF)和左室短轴缩短率(FS)降低,左室舒张末期内径(LVIDd)和左心室收缩末期内径(LVIDs)升高,乳酸脱氢酶(LDH)、肌酸激酶(CK)和心肌肌钙蛋白I(cTnI)升高;心肌出现明显损伤,纤维化面积升高;collagen I、collagenⅢ和TGF-β1的m RNA水平均升高,TUNEL阳性率升高;Bax m RNA水平升高,Bcl-2 m RNA水平降低,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)水平降低,丙二醛(MDA)水平升高,XIST水平升高(均P<0.05)。与DCM组比较,L-QDTM组、M-QDTM组和H-QDTM组的LVEF和FS均升高,LVIDd和LVIDs均降低,LDH、CK和cTnI均降低;心肌损伤减轻,纤维化面积降低;collagen I、collagenⅢ和TGF-β1的m RNA水平均降低,TUNEL阳性率降低;Bax m RNA水平降低,Bcl-2 m RNA水平升高,SOD和CAT水平升高,MDA水平均降低,XIST水平降低(均P<0.05)。结论:本研究表明芪丹通脉片在治疗阿霉素诱导扩张型心肌病大鼠中效果显著,其机制可能与抑制lncRNA XIST有关。
文摘目的探讨神经胶质瘤组织中长链非编码RNA XIST的表达及其与临床预后的关系。方法应用qRT-PCR法检测64例神经胶质瘤组织及其癌旁正常脑组织中XIST的表达,分析XIST的表达水平与胶质瘤患者临床病理学特征及其预后之间的相关性。结果 XIST在神经胶质瘤组织中的表达水平明显高于癌旁正常脑组织(2.88±0.73 vs 1.00±0.73,P=0.001 2)。同时,高级别胶质瘤中XIST的表达水平明显高于低级别胶质瘤中的表达(4.20±1.09 vs 2.88±0.73,P=0.0134)。XIST的表达水平与WHO分级和KPS评分密切相关,而与患者年龄、性别、肿瘤大小、肿瘤数目等均无显著相关性。Kaplan-Meier分析结果显示,XIST高表达组患者的生存率显著低于XIST低表达组患者(P=0.007)。神经胶质瘤中XIST高表达、WHO分级增高、KPS评分<80分均为影响患者预后的危险因素(P<0.05)。结论 XIST在神经胶质瘤组织的表达显著升高,并且与患者的临床预后显著相关,因此XIST可作为预测胶质瘤预后的生物标记物以及潜在的治疗靶点。
基金the National High Technology Research and Development Program of China (Grant No.2001AA213091) Natural Science Foundation of Hebei Prov-ince (Grant No.C2006001032)
文摘In somatic cell nuclear transfer (SCNT) technologies,the donor cell’s nuclei need to be epigenetically reprogrammed for embryonic development. The incomplete reprogramming of donor cell nuclei has been implicated as a primary reason for the low efficiency of SCNT. DNA methylation is a major epige-netic modification of the genome that regulates crucial aspects of genome function,including estab-lishment of genomic imprinting. In order to make sure whether the DNA methylation reprogramming is efficient in SCNT animals,we analyzed the DNA methylation status of two imprinting genes,H19 and Xist,in lungs of deceased SCNT bovines that died within 48 h of birth using bisulfite sequencing analysis. Our findings demonstrated that cloned bovines showed significantly lower DNA methylation of H19 than controls (P<0.05),and three tested CpGs sites (1,2,3) exhibited unmethylation in one cloned bovine (9C3); however,Xist showed similar DNA methylation levels between clones and con-trols,and both showed hypermethylation (96.11% and 86.67%).
基金supported by the National Natural Science Foundation of China (31790413, 31760657)。
文摘The limited knowledge of genomic noncoding and regulatory regions has restricted our ability to decipher the genetic mechanisms underlying complex traits in pigs. In this study, we characterized the spatiotemporal landscape of putative enhancers and promoters and their target genes by combining H3K27ac-targeted Ch IP-Seq and RNA-Seq in fetal(prenatal days 74–75) and adult(postnatal days 132–150) tissues(brain, liver, heart, muscle and small intestine) sampled from Asian aboriginal Bama Xiang and European highly selected Large White pigs of both sexes. We identified 101,290 H3K27ac peaks, marking 18,521promoters and 82,769 enhancers, including peaks that were active across all tissues and developmental stages(which could indicate safe harbor locus for exogenous gene insertion) and tissue-and developmental stage-specific peaks(which regulate gene pathways matching tissue-and developmental stage-specific physiological functions). We found that H3K27ac and DNA methylation in the promoter region of the XIST gene may be involved in X chromosome inactivation and demonstrated the utility of the present resource for revealing the regulatory patterns of known causal genes and prioritizing candidate causal variants for complex traits in pigs. In addition, we identified an average of 1,124 super-enhancers per sample and found that they were more likely to show tissue-specific activity than ordinary peaks. We have developed a web browser to improve the accessibility of the results(http://segtp.jxau.edu.cn/pencode/?genome=sus Scr11).
