Gene sequencing is a great way to interpret life, and high-throughput sequencing technology is a revolutionary technological innovation in gene sequencing researches. This technology is characterized by low cost and h...Gene sequencing is a great way to interpret life, and high-throughput sequencing technology is a revolutionary technological innovation in gene sequencing researches. This technology is characterized by low cost and high-throughput data. Currently, high-throughput sequencing technology has been widely applied in multi-level researches on genomics, transcriptomics and epigenomics. And it has fundamentally changed the way we approach problems in basic and translational researches and created many new possibilities. This paper presented a general description of high-throughput sequencing technology and a comprehensive review of its application with plain, concisely and precisely. In order to help researchers finish their work faster and better, promote science amateurs and understand it easier and better.展开更多
Oviductus Ranae is the dried oviduct of female Rana tem-poraria chensinensis （David）, distributed mainly in North- eastern China. Oviductus Ranae is one of the best-known and highly valued oriental foods and medicin...Oviductus Ranae is the dried oviduct of female Rana tem-poraria chensinensis （David）, distributed mainly in North- eastern China. Oviductus Ranae is one of the best-known and highly valued oriental foods and medicines. Traditional Chinese medicine holds that Oviductus Ranae can nourish yin, moisten lung and replenish the kidney essence. Meanwhile, activities of Oviductus Ranae such as anti-aging, anti-lipemic, anti-oxidation and anti-fatigue have also been demonstrated by modern phar-macological studies. Previous studies have shown that Oviductus Ranae is mainly composed of proteins, which are up to 50% or more.展开更多
With high-throughput Solexa sequencing technology,we profiled Porphyra yezoensis transcriptomes from 8 different samples. More than 1200 megabases from 13333334 quality paired-end reads were generated,which were assem...With high-throughput Solexa sequencing technology,we profiled Porphyra yezoensis transcriptomes from 8 different samples. More than 1200 megabases from 13333334 quality paired-end reads were generated,which were assembled into 31538 unigenes. Blast analysis showed that 56.7%unigenes were novel,which represented the specific genes of Porphyra and/or rhodophytes. Several hundreds of unigenes related to stress tolerance were discovered,including genes related to desiccation-(211) and high light-tolerance(31) ,flavonoid biosynthesis(10) ,reactive oxygen scavenging(48) and other stress-tolerance processes(208) ,which indicated there existed complex and diversity modes of stress tolerance in this species.A complete set of essential genes involved in the C3-(57) and C4-(44) carbon fixation pathway(except pyruvate phosphate dikinase) were discovered,which not only proved that they were actively transcribed but also clearly outlined the panoptic view of carbon fixation in Porphyra.Moreover,by statistically analyzing the types,proportions and frequencies of the interspersed repeats(TEs) and simple sequence repeats(SSRs) ,we discovered that the top three types of TEs were all retrotransposons and the trinucleotide was the absolute predominant type among SSRs,promoting our understanding of structural characteristics of the transcriptome.This study substantially improved the global view of the Porphyra genome and provided a valuable resource for future research.展开更多
OBJECTIVE: To corroborate the efficacy of Jintiange capsules(JTGs)( 金天格胶囊) in the treatment of osteoarthritis(OA) by exploring the potential mechanism of action of synovial mesenchymal stem cell exosomes(SMSC-Exo...OBJECTIVE: To corroborate the efficacy of Jintiange capsules(JTGs)( 金天格胶囊) in the treatment of osteoarthritis(OA) by exploring the potential mechanism of action of synovial mesenchymal stem cell exosomes(SMSC-Exos) and articular chondrocytes(ACs) through transcriptome sequencing(RNA-seq). METHODS: Type Ⅱ collagenase was used to induce OA in rats. The efficacy of JTGs was confirmed by macroscopic observation of articular cartilage, micro-CT observation, and safranin fast green staining. After SMSC-Exos and ACs were qualified, RNA-seq was used to screen differentially expressed mi RNAs and m RNAs. The target genes of differentially expressed mi RNAs in Synovial mesenchymal stem cells(SMSCs) were predicted based on the multi Mi R R package. The codifferentially expressed genes of SMSC-Exos and ACs were obtained by venny 2.1.0. The mi RNA-m RNA regulatory network was constructed by Cytoscape software. Based on the Omic Share platform, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis was performed on the m RNA regulated by key mi RNAs. Expression trend analysis was performed for co-differentially expressed genes. Correlation analysis was performed on micro-CT efficacy indicators, co-differentially expressed genes mRNA and miRNA. RESULTS: The efficacy of each administration group of JTGs was significant compared with the model group. SMSC-Exos and ACs were identified by their characteristics. The expression of rno-mi R-23a-3p, rnomi R-342-3p, rno-miR-146b-5p, rno-miR-501-3p, rnomiR-214-3p was down-regulated in OA pathological state, and the expression of rno-mi R-222-3p, rno-mi R-30e-3p, rno-mi R-676, and rno-miR-192-5p expression was upregulated, and the expression of all these mi RNAs was reversed after the intervention with JTGs containing serum. The co-differentially expressed genes were enriched in the interleukin 17 signaling pathway, tumor necrosis factor signaling pathway, transforming growth factor-β signaling pathway, etc. The expression trends of Ccl7, Akap12, Grem2, Egln3, Arhgdib, Ccl20, Mmp12, Pla2g2a, and Nr4a1 were significant. There was a correlation between micro-CT pharmacodynamic index, m RNA, and mi RNA. CONCLUSION: JTGs can improve the degeneration of joint cartilage and achieve the purpose of cartilage protection, which can be used for the treatment of OA. SMSCs-related mi RNA expression profiles were significantly altered after the intervention with JTGs containing serum. The 9 co-differentially expressed genes may be the key targets for the efficacy of JTGs in the treatment of OA rats, which can be used for subsequent validation.展开更多
Objective:Ovarian hyperstimulation syndrome(OHSS)is a frequent iatrogenic complication that arises during assisted reproduction and accounts for approximately 30%of all in vitro fertilization cycles.Using high-through...Objective:Ovarian hyperstimulation syndrome(OHSS)is a frequent iatrogenic complication that arises during assisted reproduction and accounts for approximately 30%of all in vitro fertilization cycles.Using high-throughput sequencing,we investigated the peripheral blood transcriptome of patients with OHSS.Methods:Peripheral blood samples were obtained from 15 patients in each of the OHSS high-risk and low-risk groups on the ovum pick-up day.Subsequently,high-throughput sequencing was used to obtain the peripheral blood transcriptomes of five patients each from the high-and low-risk groups.Bioinformatic tools were used for mRNA expression profile mapping and screening of differentially expressed genes(DEGs).Bioinformatics techniques were also implemented in the Kyoto Encyclopedia of Genes Genomes(KEGG)signal pathway,Gene Ontology(GO)function,and protein-protein interaction(PPI)network analyses of DEGs.Results:A total of 20,031 genes were identified and 148 were found to be differentially expressed(P<0.05,|log_(2)FC|>0.58),with 52 upregulated and 96 downregulated genes.GO and KEGG analyses indicated that these genes were involved in extracellular corpuscles(GO:0070062),plasma membrane(GO:0005886),extracellular regions(GO:0005576),immune system response(GO:0006955),PI3K-Akt signaling pathways(hsa04151),cell adhesion molecules(CAMs,hsa04514),focal adhesion(hsa04510),and complement and coagulation cascades(hsa04610).The PPI network and realtime fluorescence quantitative polymerase chain reaction(qPCR)verification predicted that complement C3,von Willebrand factor,and vascular cell adhesion protein 1 proteins are highly implicated in OHSS and may serve as potential biomarkers for future OHSS studies.Conclusion:Transcriptome analysis revealed several DEGs related to OHSS risk factors in the peripheral blood,indicating that these DEGs may be novel players in OHSS development.展开更多
Drug resistance via drug-resistant mutations in the human immunodeficiency virus(HIV)genome is the primary cause of antiviral therapy failure.Consequently,HIV drug resistance genotyping has become a critical approach ...Drug resistance via drug-resistant mutations in the human immunodeficiency virus(HIV)genome is the primary cause of antiviral therapy failure.Consequently,HIV drug resistance genotyping has become a critical approach in HIV prevention and control.Compared to the Sanger sequencing technology,high-throughput sequencing(HTS)technology has superior sensitivity and timeliness,with strong detection capabilities for low-frequency mutations.With the continued advancement of HTS technologies,their prominence in HIV drug resistance detection techniques has increased accordingly.This article will review the latest developments in HTS technology and its applications in HIV drug resistance testing.展开更多
Ovarian endometrioma(OE),also known as“chocolate cysts,”is a cystic mass that develops in the ovaries due to endometriosis and is a common gynecological condition characterized by the growth of endometrial tissue ou...Ovarian endometrioma(OE),also known as“chocolate cysts,”is a cystic mass that develops in the ovaries due to endometriosis and is a common gynecological condition characterized by the growth of endometrial tissue outside the uterus,leading to symptoms such as dysmenorrhea,pelvic pain,and infertility.However,the precise molecular and cellular mechanisms driving this pathophysiology remain largely unknown,posing challenges for diagnosis and treatment.Here,we employed integrated single-cell transcriptomic profiling of over 52,000 individual cells from endometrial tissues of OE patients and healthy donors and identified twelve major cell populations.We identified notable alterations in cell type-specific proportions and molecular signatures associated with OE.Notably,the activation of IGFBP5^(+) macrophages with pro-inflammatory properties,NK cell exhaustion,and aberrant proliferation of IQCG^(+) and KLF2^(+) epithelium are key features and may be the potential mechanisms underlying the pathogenesis of OE.Collectively,our data contribute to a better understanding of OE at the single cell level and may pave the way for the development of novel therapeutic strategies.展开更多
As one of the key technologies in biomedical research,DNA sequencing has not only improved its productivity with an exponential growth rate but also been applied to new areas of application over the past few years.Thi...As one of the key technologies in biomedical research,DNA sequencing has not only improved its productivity with an exponential growth rate but also been applied to new areas of application over the past few years.This is largely due to the advent of newer generations of sequencing platforms,offering ever-faster and cheaper ways to analyze sequences.In our previous review,we looked into technical characteristics of the nextgeneration sequencers and provided prospective insights into their future development.In this article,we present a brief overview of the advantages and shortcomings of key commercially available platforms with a focus on their suitability for a broad range of applications.展开更多
目的:基于转录组测序(RNA-seq)技术分析益肾散结复方抑制阿霉素肾病肾小球硬化的作用机制。方法:将40只SD大鼠随机分为4组,分别为正常组、阿霉素模型组、盐酸贝那普利对照组和益肾散结复方组(以下统称正常组、模型组、对照组和复方组),...目的:基于转录组测序(RNA-seq)技术分析益肾散结复方抑制阿霉素肾病肾小球硬化的作用机制。方法:将40只SD大鼠随机分为4组,分别为正常组、阿霉素模型组、盐酸贝那普利对照组和益肾散结复方组(以下统称正常组、模型组、对照组和复方组),连续2次尾静脉注射盐酸阿霉素制备阿霉素大鼠肾病模型,对照组和复方组分别按10mg/kg/d盐酸贝那普利和6.24g/kg/d益肾散结复方煎剂灌胃6周,其余组灌胃予等剂量生理盐水,灌胃前后对比大鼠一般情况、24小时尿蛋白定量(24h UTP)、血尿素氮(BUN)及血肌酐(SCr)指标变化,电镜下观察大鼠肾脏病理改变;同时将复苏的HBZY-1大鼠肾小球系膜细胞分为以上4组,以40μM浓度阿霉素诱导大鼠肾脏系膜细胞凋亡模型,分别对应处理各组细胞,观察大鼠肾小球系膜细胞凋亡情况。在此基础上,对正常组、模型组、复方组细胞进行RNA测序,筛选差异表达基因(DEG),对共同DEG进行基因本体论(GO)和京都基因和基因组百科全书(KEGG)富集分析。结果:动物实验研究显示益肾散结复方可改善阿霉素肾病大鼠便溏、反应迟钝、活动减少等精神状态,减少24h UTP、BUN和SCr水平( P <0.05),减少阿霉素肾病大鼠肾小球系膜区基质沉积,改善基底膜缺血、皱缩情况,从而抑制肾小球硬化程度;细胞实验研究显示:益肾散结复方可延缓阿霉素诱导的HBZY-1大鼠肾小球系膜细胞凋亡过程,从而对肾小球硬化起抑制作用。根据细胞RNA-seq结果,益肾散结复方抗阿霉素肾病肾小球硬化的共同DEG有863个,542个上调, 321个下调;GO分析显示这些DEG主要富集在生物过程、细胞成分和分子功能三个方面,其中与肾小球硬化相关的GO功能有:正向调节细胞迁移、细胞粘附、细胞外基质、整合素结合、CXCR趋化因子受体结合、趋化因子活动等;KEGG分析显示上述DEG富集在19条通路( P <0.05)中,其中细胞外基质(ECM)-受体相互作用、核转录因子κB(NF-κB)信号通路、钙信号通路与肾小球硬化关系密切。结论:益肾散结复方可通过抑制肾小球系膜细胞凋亡和减少系膜区基质沉积来抑制阿霉素肾病肾小球硬化过程,其作用可能通过正向调节肾小球系膜细胞迁移、参与有丝分裂细胞周期、促进细胞粘附等生物过程,作用于细胞外基质,发挥整合素结合、CXCR趋化因子受体结合和促进趋化因子活动等分子功能来实现,与调控ECM-受体相互作用、NF-κB信号通路、钙信号通路密切相关。展开更多
基金Supported by the National Natural Science Foundations of China(3127218631301791)
文摘Gene sequencing is a great way to interpret life, and high-throughput sequencing technology is a revolutionary technological innovation in gene sequencing researches. This technology is characterized by low cost and high-throughput data. Currently, high-throughput sequencing technology has been widely applied in multi-level researches on genomics, transcriptomics and epigenomics. And it has fundamentally changed the way we approach problems in basic and translational researches and created many new possibilities. This paper presented a general description of high-throughput sequencing technology and a comprehensive review of its application with plain, concisely and precisely. In order to help researchers finish their work faster and better, promote science amateurs and understand it easier and better.
基金supported by the National Key Technology Research and Development Program of the Ministry of Science and Technology of China (No. 2011BAI03B00)the National Science and Technology Major Project of the Ministry of Science and Technology of China (No. 2011ZX09401-305)
文摘Oviductus Ranae is the dried oviduct of female Rana tem-poraria chensinensis （David）, distributed mainly in North- eastern China. Oviductus Ranae is one of the best-known and highly valued oriental foods and medicines. Traditional Chinese medicine holds that Oviductus Ranae can nourish yin, moisten lung and replenish the kidney essence. Meanwhile, activities of Oviductus Ranae such as anti-aging, anti-lipemic, anti-oxidation and anti-fatigue have also been demonstrated by modern phar-macological studies. Previous studies have shown that Oviductus Ranae is mainly composed of proteins, which are up to 50% or more.
基金supported by the National Natural Science Foundation of China(30972247,30700621)Special Fund for Agro-scientific Research in the Public Interest(200903030)+2 种基金National High Technology Research and Development Program of China(2006AA10A402,2006AA10A413)Program for New Century Excellent Talents in University(NCET-06-0596)Promotive Research Fund for Young and Middle-aged Scientisits of Shandong Province(2008BS06002)
文摘With high-throughput Solexa sequencing technology,we profiled Porphyra yezoensis transcriptomes from 8 different samples. More than 1200 megabases from 13333334 quality paired-end reads were generated,which were assembled into 31538 unigenes. Blast analysis showed that 56.7%unigenes were novel,which represented the specific genes of Porphyra and/or rhodophytes. Several hundreds of unigenes related to stress tolerance were discovered,including genes related to desiccation-(211) and high light-tolerance(31) ,flavonoid biosynthesis(10) ,reactive oxygen scavenging(48) and other stress-tolerance processes(208) ,which indicated there existed complex and diversity modes of stress tolerance in this species.A complete set of essential genes involved in the C3-(57) and C4-(44) carbon fixation pathway(except pyruvate phosphate dikinase) were discovered,which not only proved that they were actively transcribed but also clearly outlined the panoptic view of carbon fixation in Porphyra.Moreover,by statistically analyzing the types,proportions and frequencies of the interspersed repeats(TEs) and simple sequence repeats(SSRs) ,we discovered that the top three types of TEs were all retrotransposons and the trinucleotide was the absolute predominant type among SSRs,promoting our understanding of structural characteristics of the transcriptome.This study substantially improved the global view of the Porphyra genome and provided a valuable resource for future research.
基金Shaanxi Province Key R&D Program (2022SF-238)Chinese Medicine Pharmaceutical Key Discipline of Shaanxi province (303061107)+1 种基金Discipline Innovation team Project of Shaanxi University of Chinese Medicine (2019-YL11)Shaanxi Province Key subject of pharmacy engineering of Shaanxi Provincial Traditional Chinese Medicine administration (2017001)。
文摘OBJECTIVE: To corroborate the efficacy of Jintiange capsules(JTGs)( 金天格胶囊) in the treatment of osteoarthritis(OA) by exploring the potential mechanism of action of synovial mesenchymal stem cell exosomes(SMSC-Exos) and articular chondrocytes(ACs) through transcriptome sequencing(RNA-seq). METHODS: Type Ⅱ collagenase was used to induce OA in rats. The efficacy of JTGs was confirmed by macroscopic observation of articular cartilage, micro-CT observation, and safranin fast green staining. After SMSC-Exos and ACs were qualified, RNA-seq was used to screen differentially expressed mi RNAs and m RNAs. The target genes of differentially expressed mi RNAs in Synovial mesenchymal stem cells(SMSCs) were predicted based on the multi Mi R R package. The codifferentially expressed genes of SMSC-Exos and ACs were obtained by venny 2.1.0. The mi RNA-m RNA regulatory network was constructed by Cytoscape software. Based on the Omic Share platform, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis was performed on the m RNA regulated by key mi RNAs. Expression trend analysis was performed for co-differentially expressed genes. Correlation analysis was performed on micro-CT efficacy indicators, co-differentially expressed genes mRNA and miRNA. RESULTS: The efficacy of each administration group of JTGs was significant compared with the model group. SMSC-Exos and ACs were identified by their characteristics. The expression of rno-mi R-23a-3p, rnomi R-342-3p, rno-miR-146b-5p, rno-miR-501-3p, rnomiR-214-3p was down-regulated in OA pathological state, and the expression of rno-mi R-222-3p, rno-mi R-30e-3p, rno-mi R-676, and rno-miR-192-5p expression was upregulated, and the expression of all these mi RNAs was reversed after the intervention with JTGs containing serum. The co-differentially expressed genes were enriched in the interleukin 17 signaling pathway, tumor necrosis factor signaling pathway, transforming growth factor-β signaling pathway, etc. The expression trends of Ccl7, Akap12, Grem2, Egln3, Arhgdib, Ccl20, Mmp12, Pla2g2a, and Nr4a1 were significant. There was a correlation between micro-CT pharmacodynamic index, m RNA, and mi RNA. CONCLUSION: JTGs can improve the degeneration of joint cartilage and achieve the purpose of cartilage protection, which can be used for the treatment of OA. SMSCs-related mi RNA expression profiles were significantly altered after the intervention with JTGs containing serum. The 9 co-differentially expressed genes may be the key targets for the efficacy of JTGs in the treatment of OA rats, which can be used for subsequent validation.
基金Gansu Provincial Science and Technology Department Grant(18YF1WA045,20YF8FA093,and 21JR11RA174)Gansu Health Industry Research Program(GSWSKY2018-16)
文摘Objective:Ovarian hyperstimulation syndrome(OHSS)is a frequent iatrogenic complication that arises during assisted reproduction and accounts for approximately 30%of all in vitro fertilization cycles.Using high-throughput sequencing,we investigated the peripheral blood transcriptome of patients with OHSS.Methods:Peripheral blood samples were obtained from 15 patients in each of the OHSS high-risk and low-risk groups on the ovum pick-up day.Subsequently,high-throughput sequencing was used to obtain the peripheral blood transcriptomes of five patients each from the high-and low-risk groups.Bioinformatic tools were used for mRNA expression profile mapping and screening of differentially expressed genes(DEGs).Bioinformatics techniques were also implemented in the Kyoto Encyclopedia of Genes Genomes(KEGG)signal pathway,Gene Ontology(GO)function,and protein-protein interaction(PPI)network analyses of DEGs.Results:A total of 20,031 genes were identified and 148 were found to be differentially expressed(P<0.05,|log_(2)FC|>0.58),with 52 upregulated and 96 downregulated genes.GO and KEGG analyses indicated that these genes were involved in extracellular corpuscles(GO:0070062),plasma membrane(GO:0005886),extracellular regions(GO:0005576),immune system response(GO:0006955),PI3K-Akt signaling pathways(hsa04151),cell adhesion molecules(CAMs,hsa04514),focal adhesion(hsa04510),and complement and coagulation cascades(hsa04610).The PPI network and realtime fluorescence quantitative polymerase chain reaction(qPCR)verification predicted that complement C3,von Willebrand factor,and vascular cell adhesion protein 1 proteins are highly implicated in OHSS and may serve as potential biomarkers for future OHSS studies.Conclusion:Transcriptome analysis revealed several DEGs related to OHSS risk factors in the peripheral blood,indicating that these DEGs may be novel players in OHSS development.
基金supported by the National Science and Technology Major Project(Grant 2018ZX10101002)the National Natural Science Foundation of China(Grants 81871694)the National Major Project of the State Key Laboratory of Infectious Diseases Prevention and Control(Grant 2011SKLID102).
文摘Drug resistance via drug-resistant mutations in the human immunodeficiency virus(HIV)genome is the primary cause of antiviral therapy failure.Consequently,HIV drug resistance genotyping has become a critical approach in HIV prevention and control.Compared to the Sanger sequencing technology,high-throughput sequencing(HTS)technology has superior sensitivity and timeliness,with strong detection capabilities for low-frequency mutations.With the continued advancement of HTS technologies,their prominence in HIV drug resistance detection techniques has increased accordingly.This article will review the latest developments in HTS technology and its applications in HIV drug resistance testing.
基金supported from Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences(CI2023D003,CI2021B014)the National Key Research and Development Program of China(2022YFC2303600,2020YFA0908000)+8 种基金the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTD-C-202002)the CACMS Innovation Fund(CI2023E002,CI2021A05101,CI2021A05104)the Science and Technology Foundation o f Shenzhen(JCYJ20210324115800001)the Science and Technology Foundation of Shenzhen(Shenzhen Clinical Medical Research Center for Geriatric Diseases)the Shenzhen Medical Research Fund(B2302051)the National Natural Science Foundation of China(82201786)Guangdong Basic and Applied Basic Research Foundation(2021A1515110646)Guangdong-Dongguan Joint Fund Regional Cultivation Project(2021B1515140033)Dongguan Science and Technology of Social Development Program(20211800904742,20221800905732,20221800904462).
文摘Ovarian endometrioma(OE),also known as“chocolate cysts,”is a cystic mass that develops in the ovaries due to endometriosis and is a common gynecological condition characterized by the growth of endometrial tissue outside the uterus,leading to symptoms such as dysmenorrhea,pelvic pain,and infertility.However,the precise molecular and cellular mechanisms driving this pathophysiology remain largely unknown,posing challenges for diagnosis and treatment.Here,we employed integrated single-cell transcriptomic profiling of over 52,000 individual cells from endometrial tissues of OE patients and healthy donors and identified twelve major cell populations.We identified notable alterations in cell type-specific proportions and molecular signatures associated with OE.Notably,the activation of IGFBP5^(+) macrophages with pro-inflammatory properties,NK cell exhaustion,and aberrant proliferation of IQCG^(+) and KLF2^(+) epithelium are key features and may be the potential mechanisms underlying the pathogenesis of OE.Collectively,our data contribute to a better understanding of OE at the single cell level and may pave the way for the development of novel therapeutic strategies.
基金This work was supported by the Chinese Academy of Sciences Scientific Research Equipments(Grant No.YZ200823)the Institutional Director’s Initiative Fund awarded to Jun Yu.
文摘As one of the key technologies in biomedical research,DNA sequencing has not only improved its productivity with an exponential growth rate but also been applied to new areas of application over the past few years.This is largely due to the advent of newer generations of sequencing platforms,offering ever-faster and cheaper ways to analyze sequences.In our previous review,we looked into technical characteristics of the nextgeneration sequencers and provided prospective insights into their future development.In this article,we present a brief overview of the advantages and shortcomings of key commercially available platforms with a focus on their suitability for a broad range of applications.
文摘目的:基于转录组测序(RNA-seq)技术分析益肾散结复方抑制阿霉素肾病肾小球硬化的作用机制。方法:将40只SD大鼠随机分为4组,分别为正常组、阿霉素模型组、盐酸贝那普利对照组和益肾散结复方组(以下统称正常组、模型组、对照组和复方组),连续2次尾静脉注射盐酸阿霉素制备阿霉素大鼠肾病模型,对照组和复方组分别按10mg/kg/d盐酸贝那普利和6.24g/kg/d益肾散结复方煎剂灌胃6周,其余组灌胃予等剂量生理盐水,灌胃前后对比大鼠一般情况、24小时尿蛋白定量(24h UTP)、血尿素氮(BUN)及血肌酐(SCr)指标变化,电镜下观察大鼠肾脏病理改变;同时将复苏的HBZY-1大鼠肾小球系膜细胞分为以上4组,以40μM浓度阿霉素诱导大鼠肾脏系膜细胞凋亡模型,分别对应处理各组细胞,观察大鼠肾小球系膜细胞凋亡情况。在此基础上,对正常组、模型组、复方组细胞进行RNA测序,筛选差异表达基因(DEG),对共同DEG进行基因本体论(GO)和京都基因和基因组百科全书(KEGG)富集分析。结果:动物实验研究显示益肾散结复方可改善阿霉素肾病大鼠便溏、反应迟钝、活动减少等精神状态,减少24h UTP、BUN和SCr水平( P <0.05),减少阿霉素肾病大鼠肾小球系膜区基质沉积,改善基底膜缺血、皱缩情况,从而抑制肾小球硬化程度;细胞实验研究显示:益肾散结复方可延缓阿霉素诱导的HBZY-1大鼠肾小球系膜细胞凋亡过程,从而对肾小球硬化起抑制作用。根据细胞RNA-seq结果,益肾散结复方抗阿霉素肾病肾小球硬化的共同DEG有863个,542个上调, 321个下调;GO分析显示这些DEG主要富集在生物过程、细胞成分和分子功能三个方面,其中与肾小球硬化相关的GO功能有:正向调节细胞迁移、细胞粘附、细胞外基质、整合素结合、CXCR趋化因子受体结合、趋化因子活动等;KEGG分析显示上述DEG富集在19条通路( P <0.05)中,其中细胞外基质(ECM)-受体相互作用、核转录因子κB(NF-κB)信号通路、钙信号通路与肾小球硬化关系密切。结论:益肾散结复方可通过抑制肾小球系膜细胞凋亡和减少系膜区基质沉积来抑制阿霉素肾病肾小球硬化过程,其作用可能通过正向调节肾小球系膜细胞迁移、参与有丝分裂细胞周期、促进细胞粘附等生物过程,作用于细胞外基质,发挥整合素结合、CXCR趋化因子受体结合和促进趋化因子活动等分子功能来实现,与调控ECM-受体相互作用、NF-κB信号通路、钙信号通路密切相关。
