OBJECTIVE To investigate the anit-inflammatory effect of extracts of Dendropanax dentiger(Harms)Merr and Lycopodiastrum casuarinoides(Spring)Holub on rheumatoid arthritis(RA)using adjuvant arthritis(AA)rat model and p...OBJECTIVE To investigate the anit-inflammatory effect of extracts of Dendropanax dentiger(Harms)Merr and Lycopodiastrum casuarinoides(Spring)Holub on rheumatoid arthritis(RA)using adjuvant arthritis(AA)rat model and possible mechanisms.METHODS The AA rat model of RA was induced in adult SparagueDawley(SD)rats by injecting of the adjuvant at base oftail.One-week-old male SD rats were randomly divided into the following groups:normal saline group(blank control),D.dentiger decoction group(80g·kg-1·d-1),L.Casuarinoides decoction group(80 g·kg·d-1),the total of glucoside Tripterygium(GTT)group(positive control,2 mg·kg-1·d-1).They were administered orally for 6weeks.Histopathology of tissues arthritis rats was observed by H.E staining.The volume of paw swelling was measured and the arthritis inflammation index was calculated.The expressions of tumor necrosis factor-α(TNF-α)and interlukin-1β(IL-1β)were detected by the ELISA assay.In addition,previous study has reported that plant-derived mi RNAs play a role for cross kingdom regulatory potential.Thus,we also performed RNA-seq technique to identify bioactive mi RNAs via comparative transcriptome analysis between D.dentiger and L.Casuarinoides.RESULTS Comparing with AA model group,the volume of paw swelling and the arthritis index were increased significantly in the AA rat model group(P<0.01),suggesting that the AA model rats were prepared properly.Compared with the AA model group,the volume of paw swelling of D.dentiger decoction group,L.Casuarinoides decoction group was decreased by 25.2%and 10.3%,respectively,and the arthritis index was decreased by 27.2%and 18.3%,respectively.Compared with AA model group,TNF-αprotein expression of D.dentiger decoction group and L.Casuarinoides decoction groups were decreased by 16.3%and 14.7%,and IL-1βprotein expression was decreased by 23.6%,18.9%(P<0.05,P<0.01),respectively.Besides,we found that some plant-derived homologous mi RNAs(such as mi RNA192 and mi RNA30a)associated with cell apoptosis processing have been screened out via comparative transcriptome analysis.But the underlying mechanisms about two mi RNAs function needs much more investigate.CONCLUSION Results showed significant anti-inflammatory effect of aqueous extracts of D.dentiger and L.Cauarinoides and justifying their therapeutic role in inflammatory condition.Furthermore,anti-inflammatory effect of D.dentiger and L.Cauarinoides may be attribute to the herb-derived mi RNAs cross-kingdom regulation.展开更多
OBJECTIVE o facilitate the basic acquaintance with the bioactive lycodine-type alkaloids biosynthetic pathways,we conducted the transcriptome analysis of L.casuarinoides by illumina sequencing.METHODS The plant of L.c...OBJECTIVE o facilitate the basic acquaintance with the bioactive lycodine-type alkaloids biosynthetic pathways,we conducted the transcriptome analysis of L.casuarinoides by illumina sequencing.METHODS The plant of L.casuarinoides was collected and was subjected to RNA isolation,cDNA library construction,and illumina sequencing before bioinformatics analysis.After sequencing,the clean reads were obtained for de novo assembly by using Trinity software,and then further processed with TGICL sequencing clustering software to generate unigenes,The unigenes are aligned by Blast X alignment to six public protein database.In addition,all unigenes are functionally annotated by GO,KEGG and characterized putative genes involved in lycopodium alkaloids biosynthesis.RESULTS In total,124,524 high-quality unigenes were obtained with an average sequence length of 601 bp.Among the L.casuarinoides transcripts,61,304 showed significant similarity(E-value<1 e-5) to the known proteins in the public database.Among the total 124 524 unigenes,47,538 open reading frame(ORFs) were predicted.Based on the bioinformatics analysis,all possible enzymes involved in the Lycodine-type alkaloids biosynthetic pathway of L.casuarinoides were identified,including primary amine oxidase(PAO),and Malonly-CoA decarboxylase.In addition,a total of 64 putative cytochrome P450(CYP450) and 827 transcription factors were selected as the candidates of Lycodine-type alka.loids modifiers.Furthermore,a total of 13 352 simple sequence repeats(SSRs) were identified from the 124,524 unigenes,of which dinucleotide motifs AG/CT(50.1%),were the most abundant.CONCLU.SION This transcriptome analysis of L.casuarinoides,provides many valuable candidate genes involv.ing in the biosynthesis of novel secondary metabolites but also lays the foundation for genetic diversity analysis via SSRs markers in L.casuarinoides.展开更多
Objective:Lycopodiastrum casuarinoides,a fern of the Lycopodiaceae family,is a traditional Chinese medicine,which has similar efficacy to that of Huperzia serrata in treating rheumatoid arthritis(RA).However,they are ...Objective:Lycopodiastrum casuarinoides,a fern of the Lycopodiaceae family,is a traditional Chinese medicine,which has similar efficacy to that of Huperzia serrata in treating rheumatoid arthritis(RA).However,they are different in the contents and compositions of lycopodium alkaloids.In this study,the biosynthesis related genes of lycopodium alkaloids and genetic markers are discovered in L.casuarinoides transcriptome.Methods:The plant of L.casuarinoides was collected and was subjected to the RNA isolation,c DNA library construction,high throughput RNA sequencing and bioinformatics analysis.Results:Totally 124,524 high-quality unigenes were assembled from RNA sequencing reads,with an average sequence length of 601 bp.Among the L.casuarinoides transcripts,61,304 shared the significant similarity(E-value<10-5)with existing protein sequences in the public databases.From 124,524 unigenes,47,538 open reading frames(ORFs)were predicted.Based on the bioinformatics analysis,all possible enzyme genes involved in the lycodine-type alkaloids biosynthetic pathway of L.casuarinoides were identified,including lysine decarboxylase(LDC),primary amine oxidase(PAO),malonyl-Co A decarboxylase,etc.Sixty-four putative cytochrome p450(CYP)and 827 putative transcription factors were selected from the transcriptome unigenes as the candidates of lycodine-type alkaloids biosynthesis modifiers.Furthermore,13,352 simple sequence repeats(SSRs)were identified from 124,524 unigenes,of which dinucleotide motifs AG/CT were the most abundant(50.1%).Meanwhile,we confirmed the amplification effectiveness of25 PCR primer pairs for randomly selected SSRs.Conclusion:We obtained the comprehensive transcriptomic information from the high throughput RNA sequencing and bioinformatics analysis,which provided a valuable resource of transcript sequences of L.casuarinoides in public databases.展开更多
基金The project supported by Science and Technology External Cooperation Key Foundation of Jiangxi Province(20151BDH80020)
文摘OBJECTIVE To investigate the anit-inflammatory effect of extracts of Dendropanax dentiger(Harms)Merr and Lycopodiastrum casuarinoides(Spring)Holub on rheumatoid arthritis(RA)using adjuvant arthritis(AA)rat model and possible mechanisms.METHODS The AA rat model of RA was induced in adult SparagueDawley(SD)rats by injecting of the adjuvant at base oftail.One-week-old male SD rats were randomly divided into the following groups:normal saline group(blank control),D.dentiger decoction group(80g·kg-1·d-1),L.Casuarinoides decoction group(80 g·kg·d-1),the total of glucoside Tripterygium(GTT)group(positive control,2 mg·kg-1·d-1).They were administered orally for 6weeks.Histopathology of tissues arthritis rats was observed by H.E staining.The volume of paw swelling was measured and the arthritis inflammation index was calculated.The expressions of tumor necrosis factor-α(TNF-α)and interlukin-1β(IL-1β)were detected by the ELISA assay.In addition,previous study has reported that plant-derived mi RNAs play a role for cross kingdom regulatory potential.Thus,we also performed RNA-seq technique to identify bioactive mi RNAs via comparative transcriptome analysis between D.dentiger and L.Casuarinoides.RESULTS Comparing with AA model group,the volume of paw swelling and the arthritis index were increased significantly in the AA rat model group(P<0.01),suggesting that the AA model rats were prepared properly.Compared with the AA model group,the volume of paw swelling of D.dentiger decoction group,L.Casuarinoides decoction group was decreased by 25.2%and 10.3%,respectively,and the arthritis index was decreased by 27.2%and 18.3%,respectively.Compared with AA model group,TNF-αprotein expression of D.dentiger decoction group and L.Casuarinoides decoction groups were decreased by 16.3%and 14.7%,and IL-1βprotein expression was decreased by 23.6%,18.9%(P<0.05,P<0.01),respectively.Besides,we found that some plant-derived homologous mi RNAs(such as mi RNA192 and mi RNA30a)associated with cell apoptosis processing have been screened out via comparative transcriptome analysis.But the underlying mechanisms about two mi RNAs function needs much more investigate.CONCLUSION Results showed significant anti-inflammatory effect of aqueous extracts of D.dentiger and L.Cauarinoides and justifying their therapeutic role in inflammatory condition.Furthermore,anti-inflammatory effect of D.dentiger and L.Cauarinoides may be attribute to the herb-derived mi RNAs cross-kingdom regulation.
基金supported by Jiangxi′s Major International Science and Technology Cooperation Projects(20151BDH80020)
文摘OBJECTIVE o facilitate the basic acquaintance with the bioactive lycodine-type alkaloids biosynthetic pathways,we conducted the transcriptome analysis of L.casuarinoides by illumina sequencing.METHODS The plant of L.casuarinoides was collected and was subjected to RNA isolation,cDNA library construction,and illumina sequencing before bioinformatics analysis.After sequencing,the clean reads were obtained for de novo assembly by using Trinity software,and then further processed with TGICL sequencing clustering software to generate unigenes,The unigenes are aligned by Blast X alignment to six public protein database.In addition,all unigenes are functionally annotated by GO,KEGG and characterized putative genes involved in lycopodium alkaloids biosynthesis.RESULTS In total,124,524 high-quality unigenes were obtained with an average sequence length of 601 bp.Among the L.casuarinoides transcripts,61,304 showed significant similarity(E-value<1 e-5) to the known proteins in the public database.Among the total 124 524 unigenes,47,538 open reading frame(ORFs) were predicted.Based on the bioinformatics analysis,all possible enzymes involved in the Lycodine-type alkaloids biosynthetic pathway of L.casuarinoides were identified,including primary amine oxidase(PAO),and Malonly-CoA decarboxylase.In addition,a total of 64 putative cytochrome P450(CYP450) and 827 transcription factors were selected as the candidates of Lycodine-type alka.loids modifiers.Furthermore,a total of 13 352 simple sequence repeats(SSRs) were identified from the 124,524 unigenes,of which dinucleotide motifs AG/CT(50.1%),were the most abundant.CONCLU.SION This transcriptome analysis of L.casuarinoides,provides many valuable candidate genes involv.ing in the biosynthesis of novel secondary metabolites but also lays the foundation for genetic diversity analysis via SSRs markers in L.casuarinoides.
基金supported by Jiangxi Foreign Science and Technology Cooperation Major Projects(Grant no.20151BDH80020)Science and Technology Project of Jiangxi Education Department(Grant no.GJJ180846)Jiangxi Postdoctoral Fund Project(Grant no.JX2016RC40)。
文摘Objective:Lycopodiastrum casuarinoides,a fern of the Lycopodiaceae family,is a traditional Chinese medicine,which has similar efficacy to that of Huperzia serrata in treating rheumatoid arthritis(RA).However,they are different in the contents and compositions of lycopodium alkaloids.In this study,the biosynthesis related genes of lycopodium alkaloids and genetic markers are discovered in L.casuarinoides transcriptome.Methods:The plant of L.casuarinoides was collected and was subjected to the RNA isolation,c DNA library construction,high throughput RNA sequencing and bioinformatics analysis.Results:Totally 124,524 high-quality unigenes were assembled from RNA sequencing reads,with an average sequence length of 601 bp.Among the L.casuarinoides transcripts,61,304 shared the significant similarity(E-value<10-5)with existing protein sequences in the public databases.From 124,524 unigenes,47,538 open reading frames(ORFs)were predicted.Based on the bioinformatics analysis,all possible enzyme genes involved in the lycodine-type alkaloids biosynthetic pathway of L.casuarinoides were identified,including lysine decarboxylase(LDC),primary amine oxidase(PAO),malonyl-Co A decarboxylase,etc.Sixty-four putative cytochrome p450(CYP)and 827 putative transcription factors were selected from the transcriptome unigenes as the candidates of lycodine-type alkaloids biosynthesis modifiers.Furthermore,13,352 simple sequence repeats(SSRs)were identified from 124,524 unigenes,of which dinucleotide motifs AG/CT were the most abundant(50.1%).Meanwhile,we confirmed the amplification effectiveness of25 PCR primer pairs for randomly selected SSRs.Conclusion:We obtained the comprehensive transcriptomic information from the high throughput RNA sequencing and bioinformatics analysis,which provided a valuable resource of transcript sequences of L.casuarinoides in public databases.