To elucidate the structure of SLA-3 alleles on inbred line of Wuzhishan pig (WZSP) population, we examined the partial exon 1, completed exon 2, and partial exon 3 of SLA-3 loci using the reverse transcription-polym...To elucidate the structure of SLA-3 alleles on inbred line of Wuzhishan pig (WZSP) population, we examined the partial exon 1, completed exon 2, and partial exon 3 of SLA-3 loci using the reverse transcription-polymerase chain reaction (RT- PCR) and the sequencing-based method in 32 WZSPs. According to pedigree and amplification results, PCR products of 8 WZSPs were selected to clone and sequence. Nine different nucleotide sequences were obtained. After comparing the DNA and protein sequences of the WZSPs SLA-3 alleles with the published GenBank SLA sequences, it was found that the SLA-3 alleles in WZSPs were all novel, but there were very few variations among them. Comparision of SLA-3 and HLA-A protein sequences indicated that there was more sequence homology. Meanwhile, the construction of a phylogenetic tree using the nucleotide sequences of 23 SLA-3 alleles and 1 HLA-A allele represented that the WZSP population owns its unique genetics resource. In this study, the alleles of SLA-3 on WZSP group were successfully detected and analyzed, which provided the firm basis on the genotype of SLA-3 for breeding specific haplotypes WZSPs.展开更多
目的探讨弱氧化性低密度脂蛋白(minimally modified low density lipoprotein,mmLDL)上调在体小鼠肠系膜动脉ETA受体的作用(endothelin type A receptors,ETA)并考察自噬是否参与这一过程。方法小鼠尾静脉注射mmLDL,腹腔注射ClassⅢPI3...目的探讨弱氧化性低密度脂蛋白(minimally modified low density lipoprotein,mmLDL)上调在体小鼠肠系膜动脉ETA受体的作用(endothelin type A receptors,ETA)并考察自噬是否参与这一过程。方法小鼠尾静脉注射mmLDL,腹腔注射ClassⅢPI3K自噬通路抑制剂6-氨基-3-甲基嘌呤(3-methyladenine,3-MA),探究自噬在mmLDL给药处理小鼠中的作用,微血管张力描记仪观察内皮素-1(endothelin-1,ET-1)引起的小鼠肠系膜动脉收缩量效曲线的变化,RT-PCR定量ETA受体mRNA,Western blot检测ETA受体和ClassⅢPI3K、Beclin-1、LC3-Ⅱ/Ⅰ、p62及p-NF-κB、NF-κB的蛋白水平表达。结果mmLDL引起ET-1收缩量效曲线明显增强,表现为Emax值由生理盐水(NS)组的(184.87±7.46)%上升为(319.91±20.31)%(P<0.001),pEC50值由NS组的(8.05±0.05)上升为(9.11±0.09)(P<0.01)。mmLDL在上调ClassⅢPI3K、beclin-1、LC3-Ⅱ/Ⅰ和下调p62蛋白水平的同时,也引起ETA受体mRNA水平、蛋白表达明显增加,增加了p-NF-κB的蛋白水平;腹腔注射3-MA抑制了mmLDL的这些作用。结论mmLDL能通过ClassⅢPI3K/Beclin-1通路激活自噬及下游NF-κB通路上调ETA受体。展开更多
文摘To elucidate the structure of SLA-3 alleles on inbred line of Wuzhishan pig (WZSP) population, we examined the partial exon 1, completed exon 2, and partial exon 3 of SLA-3 loci using the reverse transcription-polymerase chain reaction (RT- PCR) and the sequencing-based method in 32 WZSPs. According to pedigree and amplification results, PCR products of 8 WZSPs were selected to clone and sequence. Nine different nucleotide sequences were obtained. After comparing the DNA and protein sequences of the WZSPs SLA-3 alleles with the published GenBank SLA sequences, it was found that the SLA-3 alleles in WZSPs were all novel, but there were very few variations among them. Comparision of SLA-3 and HLA-A protein sequences indicated that there was more sequence homology. Meanwhile, the construction of a phylogenetic tree using the nucleotide sequences of 23 SLA-3 alleles and 1 HLA-A allele represented that the WZSP population owns its unique genetics resource. In this study, the alleles of SLA-3 on WZSP group were successfully detected and analyzed, which provided the firm basis on the genotype of SLA-3 for breeding specific haplotypes WZSPs.
文摘目的探讨弱氧化性低密度脂蛋白(minimally modified low density lipoprotein,mmLDL)上调在体小鼠肠系膜动脉ETA受体的作用(endothelin type A receptors,ETA)并考察自噬是否参与这一过程。方法小鼠尾静脉注射mmLDL,腹腔注射ClassⅢPI3K自噬通路抑制剂6-氨基-3-甲基嘌呤(3-methyladenine,3-MA),探究自噬在mmLDL给药处理小鼠中的作用,微血管张力描记仪观察内皮素-1(endothelin-1,ET-1)引起的小鼠肠系膜动脉收缩量效曲线的变化,RT-PCR定量ETA受体mRNA,Western blot检测ETA受体和ClassⅢPI3K、Beclin-1、LC3-Ⅱ/Ⅰ、p62及p-NF-κB、NF-κB的蛋白水平表达。结果mmLDL引起ET-1收缩量效曲线明显增强,表现为Emax值由生理盐水(NS)组的(184.87±7.46)%上升为(319.91±20.31)%(P<0.001),pEC50值由NS组的(8.05±0.05)上升为(9.11±0.09)(P<0.01)。mmLDL在上调ClassⅢPI3K、beclin-1、LC3-Ⅱ/Ⅰ和下调p62蛋白水平的同时,也引起ETA受体mRNA水平、蛋白表达明显增加,增加了p-NF-κB的蛋白水平;腹腔注射3-MA抑制了mmLDL的这些作用。结论mmLDL能通过ClassⅢPI3K/Beclin-1通路激活自噬及下游NF-κB通路上调ETA受体。