目的:观察六味地黄丸方对秀丽隐杆线虫寿命、生殖力、运动能力的影响并探讨其可能的分子机制。方法:秀丽隐杆线虫分为空白对照组、六味地黄丸方高、中、低剂量组(160、80、40 mg·m L-1)。寿命实验观察各组线虫平均寿命;生殖能力测...目的:观察六味地黄丸方对秀丽隐杆线虫寿命、生殖力、运动能力的影响并探讨其可能的分子机制。方法:秀丽隐杆线虫分为空白对照组、六味地黄丸方高、中、低剂量组(160、80、40 mg·m L-1)。寿命实验观察各组线虫平均寿命;生殖能力测试中,计数各组总子代数目;运动能力实验测试各组线虫20 s内身体弯曲的次数。实时荧光定量PCR(Real-Time PCR)方法检测空白组和六味地黄丸方中剂量组的daf-2 m RNA、daf-16 m RNA表达变化。结果:与空白组比较,六味地黄丸方中、低剂量组均能延长线虫的平均寿命(P<0.01),以中剂量组更为显著,并可明显提高线虫生殖高峰期及末期的生殖能力。与空白组比较,各剂量组均使线虫运动能力明显减缓(P<0.01);用药组daf-2 m RNA表达明显下调(P<0.05);daf-16 m RNA表达明显上调(P<0.05)。结论:六味地黄丸方能明显延长秀丽隐杆线虫寿命,其机制可能与下调daf-2基因和上调daf-16基因表达有关。展开更多
Objective::In order to study the important role and molecular mechanism of Brevinin-2 family antimicrobial peptide Brevinin-2ISb in methicillin-resistant Staphylococcus aureus(MRSA)infection of Caenorhabditis(C.)elega...Objective::In order to study the important role and molecular mechanism of Brevinin-2 family antimicrobial peptide Brevinin-2ISb in methicillin-resistant Staphylococcus aureus(MRSA)infection of Caenorhabditis(C.)elegans,and to find the optimal therapeutic concentration of Brevinin-2ISb.Methods::By using a C.elegans model and MRSA infection modelto study the therapeutic effect of different concentrations of Brevinin-2ISb on C.elegans.Real-time PCR was used for investigating the effect of Brevinin-2ISb on the downstream gene expression of DAF-2/DAF-16 innate immune pathway and the major virulence factor gene expression of MRSA.With protein activity tests to study the inhibitory effect of Brevinin-2ISb on MRSA virulence factor protein activity.Finally,laser confocal imaging was carried out to observe real-time expression and distribution of downstream antimicrobial proteins to further prove the effect of Brevinin-2ISb on the activation of DAF-2/DAF-16 pathway by in vivo imaging.All animal study procedures were approved by the Academic Committee at Xidian University and Xi’an Jiaotong University Animal Care and Use Committee,China(approval No.JGC201207)on July 15,2017.Results::Host immunity was largely enhanced by Brevinin-2ISb,and the expression of staphylococcal enterotoxin genes,as well as virulence factors,was suppressed by Brevinin-2ISb.Indeed,the expression of many C.elegans innate immune genes,including lys-7,spp-1,K05D8.5 and C29F3.7,was induced by Brevinin-2ISb.In particular,robust,sustained expression of the antibacterial gene lys-7 was observed after Brevinin-2ISb treatment,resulting in increased protein levels.These effects correlated with a reduction in the MRSA-mediated death of the C.elegans host.Low concentrations of Brevinin-2ISb exhibited very low hemolytic activity,and may play a positive role in host innate immunity.Specifically,activation of the DAF-2/DAF-16 pathway appears to be essential for immune activation in C.elegans treated with Brevinin-2ISb.Based on the evolutionary conservation of innate immune pathways,our results suggest that Brevinin-2ISb not only has strong antibacterial activity,but may also enhance the innate immune response in humans.This study demonstrates that Brevinin-2ISb-related peptides are potential candidates for the development of novel anti-inflammatory or anti-microbial drugs.Conclusion::Antimicrobial peptide Brevinin-2ISb effectively inhibits MRSA at low concentration.This antimicrobial peptide can prolong the life of MRSA-infected C.elegans,has very low hemolytic activity and inhibits the activity and expression of various MRSA virulence factors.More importantly,Brevinin-2ISb activated the expression of antimicrobial genes downstream of DAF-2/DAF-16,which enhanced the MRSA resistance of C.elegans.This peptide could be used as the basis for developing new drugs to replace antibiotics.展开更多
MicroRNAs(miRNAs) are short non-coding RNAs that are involved in the post-transcriptional regulation of protein-coding genes. miRNAs modulate lifespan and the aging process in a variety of organisms. In this study, we...MicroRNAs(miRNAs) are short non-coding RNAs that are involved in the post-transcriptional regulation of protein-coding genes. miRNAs modulate lifespan and the aging process in a variety of organisms. In this study, we identified a role of miR-83 in regulating lifespan of Caenorhabditis elegans. mir-83 mutants exhibited extended lifespan, and the overexpression of miR-83 was sufficient to decrease the prolonged lifespan of the mutants. We observed upregulation of the expression levels of a set of miR-83 target genes in young mir-83 mutant adults; while different sets of genes were upregulated in older mir-83 mutant adults. In vivo assays showed that miR-83 regulated expression of target genes including din-1,spp-9 and col-178, and we demonstrated that daf-16 and din-1 were required for the extension of lifespan in the mir-83 mutants. The regulation of din-1 by miR-83 during aging resulted in the differential expression of din-1 targets such as gst-4 and gst-10. In daf-2 mutants, the expression level of miR-83 was significantly reduced compared to wild-type animals. We identified a role for miR-83 in modulating lifespan in C. elegans and provided molecular insights into its functional mechanism.展开更多
文摘目的:观察六味地黄丸方对秀丽隐杆线虫寿命、生殖力、运动能力的影响并探讨其可能的分子机制。方法:秀丽隐杆线虫分为空白对照组、六味地黄丸方高、中、低剂量组(160、80、40 mg·m L-1)。寿命实验观察各组线虫平均寿命;生殖能力测试中,计数各组总子代数目;运动能力实验测试各组线虫20 s内身体弯曲的次数。实时荧光定量PCR(Real-Time PCR)方法检测空白组和六味地黄丸方中剂量组的daf-2 m RNA、daf-16 m RNA表达变化。结果:与空白组比较,六味地黄丸方中、低剂量组均能延长线虫的平均寿命(P<0.01),以中剂量组更为显著,并可明显提高线虫生殖高峰期及末期的生殖能力。与空白组比较,各剂量组均使线虫运动能力明显减缓(P<0.01);用药组daf-2 m RNA表达明显下调(P<0.05);daf-16 m RNA表达明显上调(P<0.05)。结论:六味地黄丸方能明显延长秀丽隐杆线虫寿命,其机制可能与下调daf-2基因和上调daf-16基因表达有关。
基金supported in part by the National Key R&D Program of China(No.2018YFC0910600)the National Natural Science Foundation of China(Nos.62007026,81627807,11727813,81871397,81701853,91859109)+3 种基金the Fok Ying-Tong Education Foundation of China(No.161104)the Program for the Young Top-notch Talent of Shaanxi Province,the Research Fund for Young Star of Science and Technology in Shaanxi Province of China(No.2018KJXX-018)the Science and Technology Projects of Xi’an,China(No.201809170CX11JC12)the Natural Science Basic Research Plan in Shaanxi Province of China(No.2019JQ-201).
文摘Objective::In order to study the important role and molecular mechanism of Brevinin-2 family antimicrobial peptide Brevinin-2ISb in methicillin-resistant Staphylococcus aureus(MRSA)infection of Caenorhabditis(C.)elegans,and to find the optimal therapeutic concentration of Brevinin-2ISb.Methods::By using a C.elegans model and MRSA infection modelto study the therapeutic effect of different concentrations of Brevinin-2ISb on C.elegans.Real-time PCR was used for investigating the effect of Brevinin-2ISb on the downstream gene expression of DAF-2/DAF-16 innate immune pathway and the major virulence factor gene expression of MRSA.With protein activity tests to study the inhibitory effect of Brevinin-2ISb on MRSA virulence factor protein activity.Finally,laser confocal imaging was carried out to observe real-time expression and distribution of downstream antimicrobial proteins to further prove the effect of Brevinin-2ISb on the activation of DAF-2/DAF-16 pathway by in vivo imaging.All animal study procedures were approved by the Academic Committee at Xidian University and Xi’an Jiaotong University Animal Care and Use Committee,China(approval No.JGC201207)on July 15,2017.Results::Host immunity was largely enhanced by Brevinin-2ISb,and the expression of staphylococcal enterotoxin genes,as well as virulence factors,was suppressed by Brevinin-2ISb.Indeed,the expression of many C.elegans innate immune genes,including lys-7,spp-1,K05D8.5 and C29F3.7,was induced by Brevinin-2ISb.In particular,robust,sustained expression of the antibacterial gene lys-7 was observed after Brevinin-2ISb treatment,resulting in increased protein levels.These effects correlated with a reduction in the MRSA-mediated death of the C.elegans host.Low concentrations of Brevinin-2ISb exhibited very low hemolytic activity,and may play a positive role in host innate immunity.Specifically,activation of the DAF-2/DAF-16 pathway appears to be essential for immune activation in C.elegans treated with Brevinin-2ISb.Based on the evolutionary conservation of innate immune pathways,our results suggest that Brevinin-2ISb not only has strong antibacterial activity,but may also enhance the innate immune response in humans.This study demonstrates that Brevinin-2ISb-related peptides are potential candidates for the development of novel anti-inflammatory or anti-microbial drugs.Conclusion::Antimicrobial peptide Brevinin-2ISb effectively inhibits MRSA at low concentration.This antimicrobial peptide can prolong the life of MRSA-infected C.elegans,has very low hemolytic activity and inhibits the activity and expression of various MRSA virulence factors.More importantly,Brevinin-2ISb activated the expression of antimicrobial genes downstream of DAF-2/DAF-16,which enhanced the MRSA resistance of C.elegans.This peptide could be used as the basis for developing new drugs to replace antibiotics.
基金supported by the CAS-TWAS President's Fellowshipsupported by the National Basic Research Program of China (2015CB943000)+3 种基金the National Natural Science Foundation of China (3172500146 and 31471225)the Open Project of the Chinese Academy of Sciences Key Laboratory of Innate Immunity and Chronic Disease (KLIICD-201603)the Pilot Project of Hefei Centre of Materials Science and Technology (2016FXCX006)the Strategic Priority Research Program (Pilot study) "Biological basis of aging and therapeutic strategies" of the Chinese Academy of Sciences (XDPB10)
文摘MicroRNAs(miRNAs) are short non-coding RNAs that are involved in the post-transcriptional regulation of protein-coding genes. miRNAs modulate lifespan and the aging process in a variety of organisms. In this study, we identified a role of miR-83 in regulating lifespan of Caenorhabditis elegans. mir-83 mutants exhibited extended lifespan, and the overexpression of miR-83 was sufficient to decrease the prolonged lifespan of the mutants. We observed upregulation of the expression levels of a set of miR-83 target genes in young mir-83 mutant adults; while different sets of genes were upregulated in older mir-83 mutant adults. In vivo assays showed that miR-83 regulated expression of target genes including din-1,spp-9 and col-178, and we demonstrated that daf-16 and din-1 were required for the extension of lifespan in the mir-83 mutants. The regulation of din-1 by miR-83 during aging resulted in the differential expression of din-1 targets such as gst-4 and gst-10. In daf-2 mutants, the expression level of miR-83 was significantly reduced compared to wild-type animals. We identified a role for miR-83 in modulating lifespan in C. elegans and provided molecular insights into its functional mechanism.
