Analysis of the autophagy gene expression profile of pancreatic cancer based on autophagy-related protein microtubule-associated protein 1A/1B-light chain 3

BACKGROUND Pancreatic cancer is a highly invasive malignant tumor. Expression levels of the autophagy-related protein microtubule-associated protein 1 A/1 B-light chain 3(LC3) and perineural invasion(PNI) are closely related to its occurrence and development. Our previous results showed that the high expression of LC3 was positively correlated with PNI in the patients with pancreatic cancer. In this study, we further searched for differential genes involved in autophagy of pancreatic cancer by gene expression profiling and analyzed their biological functions in pancreatic cancer, which provides a theoretical basis for elucidating the pathophysiological mechanism of autophagy in pancreatic cancer and PNI.AIM To identify differentially expressed genes involved in pancreatic cancer autophagy and explore the pathogenesis at the molecular level.METHODS Two sets of gene expression profiles of pancreatic cancer/normal tissue(GSE16515 and GSE15471) were collected from the Gene Expression Omnibus.Significance analysis of microarrays algorithm was used to screen differentially expressed genes related to pancreatic cancer. Gene Ontology(GO) analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis were used to analyze the functional enrichment of the differentially expressed genes. Protein interaction data containing only differentially expressed genes was downloaded from String database and screened. Module mining was carried out by Cytoscape software and ClusterOne plug-in. The interaction relationship between the modules was analyzed and the pivot nodes between the functional modules were determined according to the information of the functional modules and the data of reliable protein interaction network.RESULTS Based on the above two data sets of pancreatic tissue total gene expression, 6098 and 12928 differentially expressed genes were obtained by analysis of genes with higher phenotypic correlation. After extracting the intersection of the two differential gene sets, 4870 genes were determined. GO analysis showed that 14 significant functional items including negative regulation of protein ubiquitination were closely related to autophagy. A total of 986 differentially expressed genes were enriched in these functional items. After eliminating the autophagy related genes of human cancer cells which had been defined, 347 differentially expressed genes were obtained. KEGG pathway analysis showed that the pathways hsa04144 and hsa04020 were related to autophagy. In addition,65 clustering modules were screened after the protein interaction network was constructed based on String database, and module 32 contains the LC3 gene,which interacts with multiple autophagy-related genes. Moreover, ubiquitin C acts as a pivot node in functional modules to connect multiple modules related to pancreatic cancer and autophagy.CONCLUSION Three hundred and forty-seven genes associated with autophagy in human pancreatic cancer were concentrated, and a key gene ubiquitin C which is closely related to the occurrence of PNI was determined, suggesting that LC3 may influence the PNI and prognosis of pancreatic cancer through ubiquitin C.

海南黑山羊ATG16L2基因启动子区多态性研究

旨在研究海南黑山羊ATG16L2基因启动子区的结构特征及其遗传分布情况,为进一步探索该基因的表达调控机制及功能提供理论依据。本研究以200头海南黑山羊为研究对象,构建DNA混池,采用Sanger法测序对海南黑山羊ATG16L2基因启动子区的多态性进行初筛,应用PCR-RFLP技术对200头海南黑山羊个体进行基因型鉴定。对筛选到的SNP位点进行连锁不平衡分析,构建单倍型。利用生物信息学方法分析SNP位点对海南黑山羊ATG16L2基因表达的影响。在海南黑山羊ATG16L2基因启动子区共检测到3个SNPs位点,分别为SNP1(g.30667970T>C)、SNP2(g.30668540T>C)和SNP3(g.30668664C>T),且彼此连锁。SNP1和SNP2位点均表现为中度多态性,SNP3位点表现为低度多态性,且符合Hardy-Weinberg平衡(P>0.05)。单倍型分析结果显示,H1、H2、H3和H4单倍型频率分别为0.321、0.304、0.271和0.097,且H1(CGC)为优势单倍型。生物信息学分析显示,山羊ATG16L2基因共预测到3个启动子和4个CpG岛区域;存在2个重复元件LINE2(-1989~-1826 bp、-562~-426 bp)、hAT-Charlie(-1804~-1511 bp)以及5个CCAAT-Box、13个CAAT-Box、10个CGCG-Box、11个GATA-Box和2个TATA-Box。综合多种在线软件预测发现,上述SNPs可能通过影响ATG16L2基因的启动子区的顺式作用元件,从而影响海南黑山羊ATG16L2基因的转录表达。本研究在海南黑山羊ATG16L2基因启动子序列中发现3个SNPs位点,其中SNP1和SNP2表现为中度多态性,SNP3表现为低度多态性,并预测这些SNPs可能影响转录因子结合,从而调控基因表达,为进一步探究ATG16L2基因功能及其调控机制提供了理论依据。

Evaluation of autophagy-related genes and lncRNAs signature for prognositic prediction in thyroid carcinoma via bioinformatics analysis

Autophagy plays a significant role in the pathogenesis and prognosis of thyroid carcinoma.The role of autophagy-related genes and long non-coding RNAs,as well as the risk model of thyroid carcinoma patients were investigated to predict clinical outcome of thyroid carcinoma.Different expression of autophagy-related genes and long non-coding RNAs in thyroid carcinoma patients was identified in The Cancer Genome Atlas database.Functional enrichment analysis and gene set enrichment analysis was used to hint the mechanism that autophagy might act in thyroid carcinoma.Univariate and multivariate Cox regression analyses were performed for screening the prognostic autophagy-related genes and long non-coding RNAs to construct prognostic related risk model.thyroid carcinoma patients were divided into the low-risk and high-risk groups.The overall survival time was both shorter in the high-risk groups than that in the low-risk groups.As for autophagy-related genes prognostic risk model,age and autophagy-related genes risk score are independent prognostic factors that affect the survival of thyroid carcinoma.ATIC and CDKN2A expression was closely related to pathological stage and T status,DNAJB1 expression was closely related to M status,age and gender.While autophagy-associated long non-coding RNA related prognostic risk model consequently demonstrated that the long non-coding RNA risk score could significantly predict the survival rate of thyroid carcinoma patients with areas under the curve of 0.972.gene set enrichment analysis presented that a total of 16 gene sets including 10 up-regulated and 6 down-regulated gene sets were significantly enriched.The autophagy-related genes and long non-coding RNAs based prognostic risk models are a reliable forecasting tool for thyroid carcinoma patients.

ATG3 rs2638029、rs3732817基因多态性与广西地区抗中性粒细胞胞质抗体相关性小血管炎的相关性

目的探讨广西地区人群自噬相关基因(ATG)3 rs2638029、rs3732817位点多态性与原发抗中性粒细胞胞质抗体相关性小血管炎(AAV)的关联关系。方法采用多重PCR结合高通量测序技术检测194例AAV患者(AAV组)和195例健康成人(对照组)ATG3 rs2638029、ATG3 rs3732817位点的基因型,并收集受试者相关临床资料,分析其基因多态性与AAV易感性、临床症状的关系。结果ATG3 rs2638029、ATG3 rs3732817等位基因和基因型在AAV组和对照组分布差异无统计学意义(P>0.05),两位点存在连锁不平衡(D′=0.969,r^(2)=0.249),两位点可构建3个常见单体型,在AAV组和对照组中,各单体型分布差异无统计学意义(P>0.05)。ATG3 rs2638029及rs3732817不同基因型之间,水肿、血尿、蛋白尿症状发生率比较差异无统计学意义(P>0.05)。ATG3 rs2638029位点不同基因型IgG水平分布差异有统计学意义(P<0.05)。ATG3 rs3732817位点不同基因型白蛋白水平分布差异有统计学意义(P<0.05)。结论广西人群中的AAV患者ATG3基因多态性可能与IgG水平升高、低白蛋白血症相关联,但可能与AAV患者的遗传易感性无明显关联性。

Deficiency of Autophagy-Related Gene 5 in Keratinocytes Leads to Aggravation of Epidermal Damage in 2,4-Dinitrochlorobenzene-Induced Allergic Contact Dermatitis

Objective:The interrelationship between apoptosis and autophagy plays an important role in many pathophysiological processes,however,whether their interplay is involved in allergic contact dermatitis(ACD)has not yet been elucidated.So,we conducted this study to determine whether keratinocyte-specific autophagy-related gene 5(ATG5)deficiency can regulate apoptosis to inhibit skin damage in mice with 2,4-dinitrochlorobenzene(DNCB)-induced ACD.Methods:This study involved keratinocyte-specificAtg5 conditional knockout(cKO)mice(Krt14cre/+-Atg5flox/flox)and control mice(Krt14+/+-Atg5flox/flox).We painted DNCB on the right ear of each mouse to induce ACD.Dermatitis scoring and measurements of ear weight and thickness were performed to evaluate inflammation levels.An immunohistochemical assay was performed to analyze immune cell infiltration.Histological study and TUNEL staining were performed to compare the differences in skin lesions betweenAtg5 cKO mice and control mice.Immunofluorescence and western blotting were used to examine the levels of ATG5 and apoptosis-related protein.The results were statistically analyzed byt test.Results:After DNCB stimulation of mice ears,we observed a more severe phenotype inAtg5 cKO mice than in control mice(dermatitis score:7.500±2.588vs.3.250±0.822,P=0.003).Further analysis of ATG5 protein confirmed keratinocyte-specific ablation ofAtg5 in cKO mice and showed that DNCB did not influence ATG5 expression.Immunohistochemistry assay revealed that the infiltrated immune cells were not involved in aggravation of the phenotype of DNCB-stimulatedAtg5 cKO mice.However,the histological study(P=0.024),TUNEL staining(P=0.024),immunofluorescence(P=0.036),and western blotting showed that the increase in keratinocyte death,especially apoptosis,contributed to aggravation of the phenotype of DNCB-stimulatedAtg5 cKO mice.Conclusion:Deficiency ofAtg5 in keratinocytes increases apoptosis,aggravating skin damage in DNCB-induced ACD mice.This has no relationship with the involvement of immune cells.

石榴ATG基因家族鉴定及其在非生物胁迫下的表达模式分析

利用生物信息学方法,从石榴(Punica granatum Linn.)品种‘突尼斯’(‘Tunisia’)基因组中鉴定自噬相关基因(ATG)家族成员,并对其蛋白质理化性质、染色体定位、基因结构、系统进化、共线关系和表达模式进行了分析。结果表明:从石榴基因组中共鉴定出58个ATG基因,定位在8条染色体上。58个石榴ATG家庭成员可分为6个亚族。58个石榴ATG基因中3对基因存在共线关系;石榴与拟南芥[Arabidopsis thaliana(Linn.)Heynh.]和葡萄(Vitis vinifera Linn.)ATG基因家族成员间分别存在45和41对共线关系。转录组数据挖掘显示:PgVTI12b-1、PgATG1b-1、PgATG8h-1、PgATG8c-2和PgATG8c-35个基因在ABA缓解干旱胁迫的响应中差异表达;NaCl胁迫下,石榴根和叶中分别有11和8个ATG基因差异表达,且PgATG6b、PgTORb和PgATG1b-2基因在根和叶中均差异表达。实时荧光定量PCR验证发现石榴ATG基因的表达模式与转录组结果基本一致。综合分析结果表明:PgVTI12b-1、PgATG1b-1、PgATG8h-1、PgATG8c-2和PgATG8c-3基因参与石榴叶对ABA缓解干旱胁迫的响应,而PgATG6b、PgTORb和PgATG1b-2基因则同时参与石榴根和叶对NaCl胁迫的响应。

Autophagy and neurodegenerative disorders

Accumulation of aberrant proteins and inclusion bodies are hallmarks in most neurodegenerative diseases. Consequently, these aggregates within neurons lead to toxic effects, overproduction of reactive oxygen species and oxidative stress. Autophagy is a significant intracellular mechanism that removes damaged organelles and misfolded proteins in order to maintain cell homeostasis. Excessive or insufficient autophagic activity in neurons leads to altered homeostasis and influences their survival rate, causing neurodegeneration. The review article provides an update of the role of autophagic process in representative chronic and acute neurodegenerative disorders.

Genetic polymorphisms of autophagy-related gene 5 (ATG5) rs473543 predict different disease-free survivals of triple-negative breast cancer patients receiving anthracycline- and/or taxane-based adjuvant chemotherapy

Background:Autophagy plays a crucial role in chemotherapy resistance of triple-negative breast cancer(TNBC).Hence,autophagy-related gene 5(ATG5),an essential molecule involved in autophagy regulation,is presumably associated with recurrence of TNBC.This study was aimed to investigate the potential influence of single-nucleotide polymorphisms in ATG5 on the disease-free survival(DFS)of early-stage TNBC patients treated with anthracycline-and/or taxane-based chemotherapy.Methods:We genotyped ATG5 SNP rs473543 in a cohort of 316 TNBC patients treated with anthracycline-and/or taxane-based chemotherapy using the sequenom’s MassARRAY system.Kaplan-Meier survival analysis and Cox proportional hazard regression analysis were used to analyze the association between ATG5 rs473543 genotypes and the clinical outcome of TNBC patients.Results:Three genotypes,AA,GA,and GG,were detected in the rs473543 of ATG5 gene.The distribution of ATG5 rs473543 genotypes was significantly different between patients with and without recurrence(P=0.024).Kaplan-Meier survival analysis showed that patients carrying A allele of ATG5 rs473543 had an increased risk of recurrence and shorter DFS compared with those carrying the variant genotype GG in rs473543(P=0.034).In addition,after adjust-ing for clinical factors,multivariate Cox regression analyses revealed that the AA/GA genotype of rs473543 was an independent predictor for DFS(hazard risk[HR],1.73;95%confidence interval[CI],1.04-2.87;P=0.034).In addition,DFS was shorter in node-negative patients with the presence of A allele(AA/GA)than in those with the absence of A allele(P=0.027).Conclusion:ATG5 rs473543 genotypes may serve as a potential marker for predicting recurrence of early-stage TNBC patients who received anthracycline-and/or taxane-based regimens as adjuvant chemotherapy.

Screening differentially expressed genes from cyclin B1 down-regulated by high-throughput gene chip

Objective: To study differentially expressed genes by silencing cyclin B1, and to sift out autophagy-related genes. Methods: Double thymidine deoxyribonucleoside blocking was used to synchronize nasopharyngeal carcinoma cell (CNE-2) to S phase, then flow cytometry was applied to test transfection efficiency. The mRNA and protein expression level of cyclin B1 was assessed by q-PCR and western blot, respectively. Differentially expressed genes were screened by high-throughput gene chip. Results: Double thymidine deoxyribonucleoside (2.5 mmol/L) blocking was used to synchronize the cell cycle to S phase. The transfection efficiency of CNE-2 cells was 85.6%. Compared with negative group,cyclin B1-siRNA treated group significantly down-regulated mRNA expression of cyclin B1 (80%) and protein level (75.3%). Totally, 2408 differentially expressed genes were found in CNE-2, including 1245 up-regulated genes and 1163 down-regulated genes. Moreover, PTEN, an autophagy-related gene, was preliminarily sifted out. Conclusions: Cyclin B1-siRNA significantly down-regulated the expression of cyclin B1 and yielded a total of 2408 differentially expressed genes, including PETN (an autophagy-related gene).

MiR-3653 blocks autophagy to inhibit epithelial-mesenchymal transition in breast cancer cells by targeting the autophagy-regulatory genes ATG12 and AMBRA1

Background:Metastasis is the main cause of tumor-associated death and mainly responsible for treatment failure of breast cancer.Autophagy accelerates tumor metastasis.In our work,we aimed to investigate the possibility of microRNAs(miRNAs)which participate in the regulation of autophagy to inhibit tumor metastasis.Methods:MiRNA array and comprehensive analysis were performed to identify miRNAs which participated in the regulation of autophagy to inhibit tumor metastasis.The expression levels of miR-3653 in breast cancer tissues and cells were detected by quantitative real-time polymerase chain reaction.In vivo and in vitro assays were conducted to determine the function of miR-3653.The target genes of miR-3653 were detected by a dual luciferase reporter activity assay and Western blot.The relationship between miR-3653 and epithelial-mesenchymal transition(EMT)was assessed by Western blot.Student’s t-test was used to analyze the difference between any two groups,and the difference among multiple groups was analyzed with one-way analysis of variance and a Bonferroni post hoc test.Results:miR-3653 was downregulated in breast cancer cells with high metastatic ability,and high expression of miR-3653 blocked autophagic flux in breast cancer cells.Clinically,low expression of miR-3653 in breast cancer tissues(0.054±0.013 vs.0.131±0.028,t=2.475,P=0.014)was positively correlated with lymph node metastasis(0.015±0.004 vs.0.078±0.020,t=2.319,P=0.023)and poor prognosis(P<0.001).miR-3653 ameliorated the malignant phenotypes of breast cancer cells,including proliferation,migration(MDA-MB-231:0.353±0.013 vs.1.000±0.038,t=16.290,P<0.001;MDA-MB-468:0.200±0.014 vs.1.000±0.043,t=17.530,P<0.001),invasion(MDA-MB-231:0.723±0.056 vs.1.000±0.035,t=4.223,P=0.013;MDA-MB-468:0.222±0.016 vs.1.000±0.019,t=31.050,P<0.001),and colony formation(MDA-MB-231:0.472±0.022 vs.1.000±0.022,t=16.620,P<0.001;MDA-MB-468:0.650±0.040 vs.1.000±0.098,t=3.297,P=0.030).The autophagy-associated genes autophagy-related gene 12(ATG12)and activating molecule in beclin 1-regulated autophagy protein 1(AMBRA1)are target genes of miR-3653.Further studies showed that miR-3653 inhibited EMT by targeting ATG12 and AMBRA1.Conclusions:Our findings suggested that miR-3653 inhibits the autophagy process by targeting ATG12 and AMBRA1,thereby inhibiting EMT,and provided a new idea and target for the metastasis of breast cancer.

Autophagy-related protein ATG5 regulates histone H2B mono-ubiquitylation by translational control of RNF20

Autophagy is an evolutionarily conserved lysosome-mediated catabolic process（Klionsky,2007）.Autophagy is believed to be essential for cell survival,especially when cells were exposed to stresses,such as nutrient starvation.

PC12细胞在缺氧条件下的自噬现象

目的:考察PC12细胞内自噬发生与缺氧时间的关系,探讨自噬对缺氧细胞的影响作用。方法:以PC12细胞为模型,将对数生长期的细胞加入96孔培养板,37℃、5%CO_2培养24 h后,放入0.5%O_2、94.5%N_2和5%CO_2的培养箱缺氧1 h、3 h、6h、9 h、12 h、24 h、36 h和48 h,用MTT法检测细胞存活率,透射电镜观察细胞内自噬体,Westen blot法检测自噬相关蛋白(LC3B、Atg5和Beclin1)的表达,用试剂盒检测细胞内乳酸脱氢酶(LDH)活性、活性氧自由基(ROS)和线粒体膜电位(MNP)水平,探究缺氧不同时间自噬对PC12细胞的作用。结果:在缺氧3~12 h,PC12细胞自噬明显增加,自噬相关蛋白(LC3B、Atg5和Beclin1)表达增加,尤其是缺氧9 h,PC12细胞存活明显增加,表明短时间缺氧自噬对细胞起保护作用,而随着缺氧时间的延长细胞存活明显降低,自噬相关蛋白表达水平减少,细胞LHD和ROS水平明显增加。MMP水平显著下降,细胞凋亡明显增加。结论:在缺氧早期自噬对PC12细胞起保护作用,但缺氧时间较长,超过了细胞自身调节能力导致细胞死亡。

猪瘟病毒促进细胞自噬并利于病毒增殖

细胞自噬在病毒的增殖、释放中有着重要的作用,目前国内仍然没有猪瘟病毒与细胞自噬相互作用关系的报道。本研究旨在探讨猪瘟病毒Shimen株感染宿主细胞(ST)后对细胞自噬的影响,以及细胞自噬对病毒增殖的作用。通过透射电镜和自噬体标记分子LC3的荧光聚点的观察,直观判断病毒对自噬的影响;并通过Western blot,检测LC3蛋白和P62蛋白的表达量,以及LC3蛋白转化分析,确定猪瘟病毒感染促进细胞自噬发生。利用自噬促进剂雷帕霉素、抑制剂3-MA、自噬体与溶酶体融合阻断剂氯喹(CQ),以及自噬基因Beclin 1、LC3蛋白的干扰,调控自噬来研究自噬对病毒增殖的影响。结果表明,病毒感染细胞促进了细胞自噬的发生,且能形成完整的自噬过程,同时病毒利用细胞自噬来促进自身增殖。本研究为探索猪瘟病毒与宿主细胞相互作用关系增加了新的数据。

自体吞噬在植物生长发育中的功能

自体吞噬是一种细胞内自我降解系统,它能将植物细胞内溶物运输至液泡并降解。自体吞噬可划分为内溶物的包裹、运输至液泡、内溶物的降解和降解产物的重新利用几个连续步骤。关于细胞自体吞噬的认识主要来源于酵母、人类、小鼠、果蝇和线虫等生物,以拟南芥等为代表的植物细胞自体吞噬的研究虽然刚刚开始,但也取得了一些标志性的成果,且近十几年来已迅速成为植物研究领域的热点之一。自体吞噬在植物体内具有多种生理和病理作用,如对饥饿的适应、细胞内蛋白质和细胞器的清除、种子中贮藏蛋白的积累、抵制微生物、细胞死亡和胁迫响应等。本文在介绍自体吞噬形成过程的基础上,着重探讨了自体吞噬在植物生长发育中的功能,并对植物中自体吞噬的研究方向进行了展望。

细胞自噬的基因调控及其与稻瘟病的关系

细胞自噬是真核生物中广泛存在的过程,并且在进化上十分保守。在真核生物分化和发育的过程中,它参与胞内细胞器和蛋白质的周转,被认为在细胞的形态建成方面发挥重要作用。现就细胞自噬的分子机制和功能做一介绍,并对稻瘟病菌细胞自噬的研究现状进行了回顾。

显微镜技术在植物细胞自噬研究中的应用

细胞自噬是真核生物对细胞内物质进行循环利用的重要途径。在植物中已发现的细胞自噬有两种形式,即微自噬和巨自噬。自噬在植物体的生长发育、免疫应答、胁迫反应、衰老等过程中具有非常重要的作用。显微镜技术是研究细胞自噬的重要手段,与生物化学和分子生物学技术相结合,荧光显微镜,激光扫描共聚焦显微镜,电子显微镜等技术的相互印证,大大地推动了对细胞自噬过程和机理的研究。本文扼要概述了显微镜技术在植物细胞自噬研究中的应用。

结核分枝杆菌ESAT6-CFP10融合蛋白对小鼠巨噬细胞自噬功能的影响

目的研究结核分枝杆菌(MTB)ESAT6-CFP10融合蛋白对小鼠巨噬细胞自噬功能的影响。方法H37Rv菌株感染小鼠巨噬细胞后加入纯化的重组ESAT6-CFP10融合蛋白,通过透射电镜检测自噬体的形成。提取细胞总RNA和蛋白,以实时定量RT-PCR及Western blot方法检测自噬相关基因(atg)分子水平和蛋白表达水平。结果ESAT6-CFP10融合蛋白可抑制小鼠巨噬细胞自噬体的形成,并导致atg分子表达水平下降,其中atg8表达量下降最为明显。结论MTB ESAT6-CFP10融合蛋白通过调控atg分子表达水平影响小鼠巨噬细胞自噬功能。

新风胶囊通过调节自噬相关蛋白的表达改善佐剂关节炎大鼠的肺功能

目的观察新风胶囊(XFC)对佐剂关节炎(AA)大鼠滑膜、肺组织自噬相关基因(ATG)、微管相关蛋白1轻链3Ⅱ(LC3-Ⅱ)和beclin 1的影响。方法将大鼠随机分为正常对照(NC)组、模型对照(MC)组、来氟米特(LEF)组、XFC组,每组10只,除NC组外均将Freund完全佐剂(0.1 m L/只)注射于每只大鼠右后足跖皮内以致炎,第10天在尾根部注射Freund完全佐剂0.05 m L加强免疫,复制AA模型。致炎后第13天给药。NC组、MC组给予生理盐水灌胃,每天1次;LEF、XFC组给予相应药物灌胃,每天1次,连续给药30 d。观察大鼠足跖肿胀度、关节炎指数、肺功能,ELISA检测血清细胞因子B细胞刺激因子(BAFF)、白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、IL-4、IL-10表达,透射电镜观察滑膜、肺组织自噬小体,反转录PCR检测大鼠滑膜、肺组织ATG5、ATG7、ATG12 mRNA,Western blot法检测大鼠滑膜、肺组织LC3-Ⅱ、beclin 1的蛋白表达。结果 XFC组大鼠肺功能参数、血清IL-4、IL-10表达升高;滑膜、肺组织自噬体及自噬溶酶体较MC组增加;血清IL-1β、BAFF表达及滑膜组织ATG7、ATG12 mRNA降低,肺组织ATG5、ATG7 mRNA降低,ATG12 mRNA升高;滑膜组织、肺组织LC3-Ⅱ、beclin 1升高。结论 XFC通过调节AA大鼠滑膜、肺组织细胞自噬,改善肺功能。

番茄红素对心肌细胞缺氧损伤的作用机制

目的探讨番茄红素对心肌细胞沉默信息调节因子1(Sirt1)-自噬通路的作用。方法原代培养乳鼠心肌细胞予以无血清、无糖培养及缺氧干预,模拟心肌缺血,用番茄红素和Sirt1的抑制剂尼克酰胺干预。采用Western blot法检测Sirt1、自噬相关基因6(Atg6)Beclin1及自噬相关基因8LC3蛋白的表达;RT-PCR技术检测心肌细胞Sirt1的mRNA表达。结果①在心肌缺氧中,番茄红素提高了细胞Sirt1蛋白和mRNA的表达水平;②番茄红素增加了心肌缺氧时自噬相关基因Beclin1和LC3蛋白的表达;③Sirt1的抑制剂尼克酰胺阻断了番茄红素对自噬蛋白的作用。结论番茄红素可能通过Sirt1激活自噬,从而减少心肌缺氧损伤。

心肌细胞缺氧中番茄红素对自噬的作用

目的探讨在心肌缺氧损伤中,番茄红素的作用及对自噬的影响。方法原代培养乳鼠心肌细胞予以无血清、无糖培养及缺氧干预,模拟心肌缺血,用番茄红素干预。在光学显微镜下观察缺氧后心肌细胞的形态;利用CCK-8法测量心肌细胞的存活率;及免疫印迹方法检测自噬相关基因LC3蛋白表达。结果 1番茄红素提高了心肌缺氧时细胞的存活率。2番茄红素增加了LC3蛋白的表达。3自噬的阻断剂3-甲基腺嘌呤(3-MA)削弱了番茄红素保护心肌细胞的作用。结论番茄红素可能通过激活自噬来提高心肌细胞抗缺血缺氧的能力。