Objective： To study the effect of the extract of Prunella vulgaris L. on multiple drugs resistant bacillus tuberculosis （MDR-TB）. Methods： Experimental animal model in rats was induced by MDR-TB. Normal group mode...Objective： To study the effect of the extract of Prunella vulgaris L. on multiple drugs resistant bacillus tuberculosis （MDR-TB）. Methods： Experimental animal model in rats was induced by MDR-TB. Normal group model group and Prunella vulgaris L. group were set up. The contents of IFN-7, IL-4, IL-10 and IL-12 were examined by ELISA. Their genome mRNAs were extracted, the target genes were amplified by PCR. RT-PCR was used to detect the mRNA levels of them. Results： The content of IFN-q, of the extract of Prunella vulgaris L. group was 1.98±0.67 pg/ml, IL-4 was 6.47±1.46 pg/ml, IL-10 was 12.13±3.43 pg/ml and IL-12 was 3.02±0.86 pg/ml. Compared with the model group, Prunella vulgaris L. group was notable difference in serum IFN-γ, IL-12 and IL-10 （P〈0.05）. The mRNA levels of IFN-γ, IL-12 increased and IL-10 decreased obviously, the differences were quite significant （P〈0.05）, but IL-4 had no obvious change. Conclusion： The extract of Prunella vulgaris L. can enhance the cellar immunological function in rats from up-regulation of the level of genetic transcription, accordingly provide the theory basis of healing of tuberculosis with it.展开更多
Objective To study the regulatory roles of SIRT1 on EZH2 expression and the further ef-fects on EZH2's repression of target gene expression. Methods The stable SIRT1 RNAi and Control RNAi HeLa cells were establish...Objective To study the regulatory roles of SIRT1 on EZH2 expression and the further ef-fects on EZH2's repression of target gene expression. Methods The stable SIRT1 RNAi and Control RNAi HeLa cells were established by in-fection with retroviruses expressing shSIRT1 and shLuc respectively followed by puromycin selection. EZH2 protein level was detected by Western blot in either whole cell lysate or the fractional cell extract. Reverse transcription-polymerase chain reaction was performed to detect the mRNA level of EZH2. Cycloheximide was used to treat SIRT1 RNAi and Control RNAi cells for protein stability assay. Chromatin immunoprecipitation (ChIP) assay was applied to measure enrichment of SIRT1, EZH2, and trimethylated H3K27 (H3K27me3) at SATB1 promoter in SIRT1 RNAi and Control RNAi cells. Results Western blot results showed that EZH2 protein level increased upon SIRT1 de-pletion. Fractional extraction results showed unchanged cytoplasmic fraction and increased chromatin fraction of EZH2 protein in SIRT1 RNAi cells. The mRNA level of EZH2 was not affected by knockdown of SIRT1. SIRT1 recruitment was not detected at the promoter region of EZH2 gene locus. The protein stability assay showed that the protein stability of EZH2 increases upon SIRT1 knockdown. Upon SIRT1 depletion, EZH2 and H3K27me3 recruitment at SATB1 promoter increases and the mRNA level of SATB1 decreases. Conclusions Depletion of SIRT1 increases the protein stability of EZH2. The regulation of EZH2 protein level by SIRT1 affects the repressive effects of EZH2 on the target gene expres-sion.展开更多
基金Supported by the Natural Science Foundation for Universities in Anhui Province (KJ2010A087 and KJ2008A152)
文摘Objective： To study the effect of the extract of Prunella vulgaris L. on multiple drugs resistant bacillus tuberculosis （MDR-TB）. Methods： Experimental animal model in rats was induced by MDR-TB. Normal group model group and Prunella vulgaris L. group were set up. The contents of IFN-7, IL-4, IL-10 and IL-12 were examined by ELISA. Their genome mRNAs were extracted, the target genes were amplified by PCR. RT-PCR was used to detect the mRNA levels of them. Results： The content of IFN-q, of the extract of Prunella vulgaris L. group was 1.98±0.67 pg/ml, IL-4 was 6.47±1.46 pg/ml, IL-10 was 12.13±3.43 pg/ml and IL-12 was 3.02±0.86 pg/ml. Compared with the model group, Prunella vulgaris L. group was notable difference in serum IFN-γ, IL-12 and IL-10 （P〈0.05）. The mRNA levels of IFN-γ, IL-12 increased and IL-10 decreased obviously, the differences were quite significant （P〈0.05）, but IL-4 had no obvious change. Conclusion： The extract of Prunella vulgaris L. can enhance the cellar immunological function in rats from up-regulation of the level of genetic transcription, accordingly provide the theory basis of healing of tuberculosis with it.
基金Supported by National Natural Science Foundation of China (30721063)National Basic Research Program of China (973 Program) (2005CB522402, 2006CB910403)+1 种基金National Laboratory of Medical Molecular Biology grant (2060204)Beijing municipal government grant (YB20081002301)
文摘Objective To study the regulatory roles of SIRT1 on EZH2 expression and the further ef-fects on EZH2's repression of target gene expression. Methods The stable SIRT1 RNAi and Control RNAi HeLa cells were established by in-fection with retroviruses expressing shSIRT1 and shLuc respectively followed by puromycin selection. EZH2 protein level was detected by Western blot in either whole cell lysate or the fractional cell extract. Reverse transcription-polymerase chain reaction was performed to detect the mRNA level of EZH2. Cycloheximide was used to treat SIRT1 RNAi and Control RNAi cells for protein stability assay. Chromatin immunoprecipitation (ChIP) assay was applied to measure enrichment of SIRT1, EZH2, and trimethylated H3K27 (H3K27me3) at SATB1 promoter in SIRT1 RNAi and Control RNAi cells. Results Western blot results showed that EZH2 protein level increased upon SIRT1 de-pletion. Fractional extraction results showed unchanged cytoplasmic fraction and increased chromatin fraction of EZH2 protein in SIRT1 RNAi cells. The mRNA level of EZH2 was not affected by knockdown of SIRT1. SIRT1 recruitment was not detected at the promoter region of EZH2 gene locus. The protein stability assay showed that the protein stability of EZH2 increases upon SIRT1 knockdown. Upon SIRT1 depletion, EZH2 and H3K27me3 recruitment at SATB1 promoter increases and the mRNA level of SATB1 decreases. Conclusions Depletion of SIRT1 increases the protein stability of EZH2. The regulation of EZH2 protein level by SIRT1 affects the repressive effects of EZH2 on the target gene expres-sion.
