The liver is a vital organ in chickens that performs a number of crucial physiological functions, including the storage of hepatic glycogen, protein synthesis, detoxification, and deoxidation. The growth and metabolis...The liver is a vital organ in chickens that performs a number of crucial physiological functions, including the storage of hepatic glycogen, protein synthesis, detoxification, and deoxidation. The growth and metabolism of the liver are complex processes influenced by factors such as environment, diet, and genetics. MicroRNAs(miRNAs), as posttranscriptional regulatory molecules, play a role in various biological processes. There is growing evidence that miR-27b-5p plays a key role in the regulation of liver development and metabolism in various species. However, its role in chicken livers has yet to be determined. In our experiment, we found that chickens with fatty livers had significantly higher levels of serum triglyceride(TG) and total cholesterol(TC) compared to the normal chickens, while the control group had significantly higher levels of very low-density lipoprotein(VLDL) and serum hormones. Further research showed that the mRNA of miR-27b-5p was highly expressed in fatty livers. By exploring the function of miR-27b-5p in chicken livers, we discovered that it promotes lipogenesis, oxidative stress, and inflammatory responses, leading to hepatocyte apoptosis. Our study also established the mechanism by which miR-27b-5p interacts with its target gene, and found that miR-27b-5p targets insulin receptor substrate 2(IRS2) and modulates the PI3K/AKT signaling pathway. Additionally, our investigation of IRS2 in chicken hepatocytes revealed that knocking down IRS2 has the same effects as overexpressing miR-27b-5p. In conclusion, our study revealed that miR-27b-5p directly binds to IRS2, inhibiting the PI3K/AKT signaling pathway and causing steatosis, oxidative stress, inflammation, and apoptosis in chicken liver.展开更多
目的探讨应用CRISPR/Cas9技术制备敲除雄激素受体(AR)和胰岛素受体底物2(IRS2)基因大鼠,构建可用于研究的稳定遗传的基因敲除大鼠品系。方法针对与雄激素表达相关基因-AR基因和胰岛素表达有关的胰岛素受体底物(IRS)基因设计出了20 bp的A...目的探讨应用CRISPR/Cas9技术制备敲除雄激素受体(AR)和胰岛素受体底物2(IRS2)基因大鼠,构建可用于研究的稳定遗传的基因敲除大鼠品系。方法针对与雄激素表达相关基因-AR基因和胰岛素表达有关的胰岛素受体底物(IRS)基因设计出了20 bp的AR/IRS2 sgRNA和30 bp AR/IRS2 sgRNA,设计出的sgRNA和Cas9体外转录后显微注射至SD大鼠受精卵细胞,F0代大鼠出生后取其基因组DNA测序鉴定基因型后,对确认敲除的大鼠F0与野生型交配,获得杂合子F1代。结果设计了20 bp AR/IRS2 sgRNA和30 bp AR/IRS2 sgRNA,与Cas9一起进行了体外转录后显微注射至SD大鼠受精卵细胞,共获得2个受精卵并移植到1只雌性代孕大鼠。繁殖出嵌合体大鼠F0 2只(#51,#56),F0#51、#56与野生型大鼠合笼,繁殖出F1大鼠9只,其中F0#51繁殖出3只为双敲大鼠(#12,#13,#18),其余6只为单敲大鼠,F0#51产生的3只双敲大鼠发生不同程度的碱基插入或缺失,其中#12,#18大鼠在AR基因上缺失7个碱基(CCCCGGC),在IRS2基因上缺失4个碱基(GCGC)。#13大鼠在AR基因上插入2个碱基(CG),在IRS2基因上缺失4个碱基(GCGC)。大鼠的AR、IRS2基因表达被破坏。结论采用CRISPR/Cas9技术成功获得敲除基因AR、IRS2大鼠,经基因测序和蛋白质免疫印迹鉴定AR、IRS2,证明正确,为下一步鉴定是否表现出多囊卵巢综合征表型特征打下基础。展开更多
目的:探讨针刺对2型糖尿病(T2DM)大鼠肝组织胰岛素受体底物1和2基因(IRS1和2mRNA)表达的影响。方法:给食源性肥胖大鼠腹腔注射小剂量链脲佐菌素(STZ)(25mg/kg)造模成T2DM大鼠,随机分为针刺组、优降糖组和模型组,处理4周后,用快速血糖仪...目的:探讨针刺对2型糖尿病(T2DM)大鼠肝组织胰岛素受体底物1和2基因(IRS1和2mRNA)表达的影响。方法:给食源性肥胖大鼠腹腔注射小剂量链脲佐菌素(STZ)(25mg/kg)造模成T2DM大鼠,随机分为针刺组、优降糖组和模型组,处理4周后,用快速血糖仪检测空腹血糖(FBS)、用放免法检测空腹胰岛素(FINS)、用实时定量荧光PCR(real-tim e RT PCR)法检测肝组织IRS1和2mRNA表达,并与正常组大鼠进行对照。结果:针刺组和优降糖组FBS和FINS比模型组显著降低(P<0.05);模型组IRS1mRNA相对含量是正常组的25%,针刺组为正常组的2.83倍,优降糖组为正常组的1.23倍。模型组IRS2mRNA相对含量为正常组的2.19倍,针刺组IRS2mRNA相对含量为正常组的4.84%,优降糖组IRS2mRNA相对含量为正常组的4.59倍。结论:肝组织IRS1和2mRNA表达异常可能是T2DM的发病机制之一,针刺和优降糖均可上调T2DM大鼠肝组织IRS1mRNA的表达,针刺对T2DM大鼠肝组织IRS1 mRNA表达的上调作用优于优降糖;针刺具有显著抑制T2DM大鼠肝组织IRS2mRNA表达的作用,而优降糖无此作用。展开更多
基金financially supported by the National Key Research and Development Program of China (2021YFD1300600)the Sichuan Science and Technology Program, China (2021YFYZ0007 and 2022YFYZ0005)the China Agriculture Research System of MOF and MARA (CARS-40-K06)。
文摘The liver is a vital organ in chickens that performs a number of crucial physiological functions, including the storage of hepatic glycogen, protein synthesis, detoxification, and deoxidation. The growth and metabolism of the liver are complex processes influenced by factors such as environment, diet, and genetics. MicroRNAs(miRNAs), as posttranscriptional regulatory molecules, play a role in various biological processes. There is growing evidence that miR-27b-5p plays a key role in the regulation of liver development and metabolism in various species. However, its role in chicken livers has yet to be determined. In our experiment, we found that chickens with fatty livers had significantly higher levels of serum triglyceride(TG) and total cholesterol(TC) compared to the normal chickens, while the control group had significantly higher levels of very low-density lipoprotein(VLDL) and serum hormones. Further research showed that the mRNA of miR-27b-5p was highly expressed in fatty livers. By exploring the function of miR-27b-5p in chicken livers, we discovered that it promotes lipogenesis, oxidative stress, and inflammatory responses, leading to hepatocyte apoptosis. Our study also established the mechanism by which miR-27b-5p interacts with its target gene, and found that miR-27b-5p targets insulin receptor substrate 2(IRS2) and modulates the PI3K/AKT signaling pathway. Additionally, our investigation of IRS2 in chicken hepatocytes revealed that knocking down IRS2 has the same effects as overexpressing miR-27b-5p. In conclusion, our study revealed that miR-27b-5p directly binds to IRS2, inhibiting the PI3K/AKT signaling pathway and causing steatosis, oxidative stress, inflammation, and apoptosis in chicken liver.
文摘目的探讨应用CRISPR/Cas9技术制备敲除雄激素受体(AR)和胰岛素受体底物2(IRS2)基因大鼠,构建可用于研究的稳定遗传的基因敲除大鼠品系。方法针对与雄激素表达相关基因-AR基因和胰岛素表达有关的胰岛素受体底物(IRS)基因设计出了20 bp的AR/IRS2 sgRNA和30 bp AR/IRS2 sgRNA,设计出的sgRNA和Cas9体外转录后显微注射至SD大鼠受精卵细胞,F0代大鼠出生后取其基因组DNA测序鉴定基因型后,对确认敲除的大鼠F0与野生型交配,获得杂合子F1代。结果设计了20 bp AR/IRS2 sgRNA和30 bp AR/IRS2 sgRNA,与Cas9一起进行了体外转录后显微注射至SD大鼠受精卵细胞,共获得2个受精卵并移植到1只雌性代孕大鼠。繁殖出嵌合体大鼠F0 2只(#51,#56),F0#51、#56与野生型大鼠合笼,繁殖出F1大鼠9只,其中F0#51繁殖出3只为双敲大鼠(#12,#13,#18),其余6只为单敲大鼠,F0#51产生的3只双敲大鼠发生不同程度的碱基插入或缺失,其中#12,#18大鼠在AR基因上缺失7个碱基(CCCCGGC),在IRS2基因上缺失4个碱基(GCGC)。#13大鼠在AR基因上插入2个碱基(CG),在IRS2基因上缺失4个碱基(GCGC)。大鼠的AR、IRS2基因表达被破坏。结论采用CRISPR/Cas9技术成功获得敲除基因AR、IRS2大鼠,经基因测序和蛋白质免疫印迹鉴定AR、IRS2,证明正确,为下一步鉴定是否表现出多囊卵巢综合征表型特征打下基础。
文摘目的:探讨针刺对2型糖尿病(T2DM)大鼠肝组织胰岛素受体底物1和2基因(IRS1和2mRNA)表达的影响。方法:给食源性肥胖大鼠腹腔注射小剂量链脲佐菌素(STZ)(25mg/kg)造模成T2DM大鼠,随机分为针刺组、优降糖组和模型组,处理4周后,用快速血糖仪检测空腹血糖(FBS)、用放免法检测空腹胰岛素(FINS)、用实时定量荧光PCR(real-tim e RT PCR)法检测肝组织IRS1和2mRNA表达,并与正常组大鼠进行对照。结果:针刺组和优降糖组FBS和FINS比模型组显著降低(P<0.05);模型组IRS1mRNA相对含量是正常组的25%,针刺组为正常组的2.83倍,优降糖组为正常组的1.23倍。模型组IRS2mRNA相对含量为正常组的2.19倍,针刺组IRS2mRNA相对含量为正常组的4.84%,优降糖组IRS2mRNA相对含量为正常组的4.59倍。结论:肝组织IRS1和2mRNA表达异常可能是T2DM的发病机制之一,针刺和优降糖均可上调T2DM大鼠肝组织IRS1mRNA的表达,针刺对T2DM大鼠肝组织IRS1 mRNA表达的上调作用优于优降糖;针刺具有显著抑制T2DM大鼠肝组织IRS2mRNA表达的作用,而优降糖无此作用。
