外泌体miR-25-3p和miR-425-5p对持续ALT正常慢性乙肝患者肝脏炎症的早期诊断价值及机制研究

批准号:
81800532
项目类别:
青年科学基金项目
资助金额:
20.0 万元
负责人:
符小玉
依托单位:
学科分类:
H0309.炎性及感染性肝病
结题年份:
2021
批准年份:
2018
项目状态:
已结题
项目参与者:
谭德明、欧阳奕、易盼盼、余松蔓、李荣华、侯帅
国基评审专家1V1指导 中标率高出同行96.8%
结合最新热点,提供专业选题建议
深度指导申报书撰写,确保创新可行
指导项目中标800+,快速提高中标率
微信扫码咨询
中文摘要
慢性乙型肝炎(CHB)危害严重,大量研究证实部分持续ALT正常(PNALT)患者存在显著肝损伤,尽早发现这类患者的肝脏损伤并给予抗病毒治疗是关键。本课题前期在PNALT的CHB 患者中通过高通量测序筛选出外泌体miR-25-3p/miR-425-5p与肝脏炎症程度密切相关,并确认了miR-25-3p的下游靶基因为肝脏炎症和免疫相关基因TLR3、TRAF6和EGR2。本研究拟首先扩大临床样本量,探讨这两个miRNAs对肝脏炎症早期诊断的可行性;接着通过荧光标记的手段研究外泌体在肝细胞中的转运,共培养外泌体与受体肝细胞,分析外泌体miRNAs对肝细胞炎症损伤和免疫信号调节的影响;最后构建慢性肝损伤小鼠模型,观察外泌体miRNAs影响肝脏炎症的机制及抗乙肝病毒的免疫治疗作用。阐明外泌体miR-25-3p/miR-425-5p对PNALT的早期肝损伤的诊断价值,为该类患者抗病毒治疗提供新思路。
英文摘要
Chronic hepatitis B (CHB) is fatal. A large number of studies have shown that there are significant liver injuries in some patients with persistent normal ALT (PNALT). Early detection of liver injury and antiviral therapy is very important. Our previous study showed that exosome-derived miR-25-3p / miR-425-5p in PNALT CHB patients by high-throughput sequencing was closely related to the degree of liver inflammation, and confirmed that the downstream target gene of miR-25-3p was liver inflammation and immune-related genes such as TLR3, TRAF6 and EGR2. In this study, we first expand the clinical sample size to explore the feasibility of these two miRNAs in the diagnosis of early liver injury; then analyzed the effects of exosome miRNAs on hepatocyte inflammatory injury and immune signal regulation followed by fluorescence labeling of exosomes in liver cell transport, co-culture of exosomes and recipient hepatocytes. Finally, constructed the chronic liver injury model mice to observe the mechanism of exogenous miRNAs affecting hepatic inflammatory injury and the anti-hepatitis B virus immunotherapy in vivo. Thereby, the study elucidate the significance of exosomal miR-25-3p/miR-425-5p as novel candidate for early liver injury and treatment of PNALT.
本项目在对慢性乙型肝炎持续肝功能正常(PNALT)患者外周血外泌体进行高通量测序及qRT-PCR验证后,发现外泌体来源的miR-25-3p表达水平与肝组织炎症分级密切相关。按照研究计划,通过细胞实验验证外泌体的功能,qRT-PCR、凋亡实验、Transwell和细胞划痕试验等,详细分析外泌体miR-25-3p的功能,通过干预miR-25-3p的表达,确认其影响肝癌细胞凋亡及增殖的作用。并验证其可靶向调控FBXW7,调节PI3K/Akt通路,另一方面,发现miR-25-3p也可调控SIK1 ,调节 Wnt/β-catenin通路。动物成瘤实验也证实外泌体miR-25-3p通过上述通路影响肝癌细胞的炎症凋亡。全面系统阐述外泌体来源的miR-25-3p可能是PNALT患者由慢性乙型肝炎发展为肝癌的潜在机制miR-25-3p可以促进肝癌的细胞转移和增殖,同时抑制它们的凋亡。.本研究一方面证实外泌体来源miR-25-3p对PNALT慢性乙肝肝脏炎症早期诊断的可靠性; 同时通过体内体外实验,详细分析外泌体分泌miR-25-3p影响肝脏炎症的作用及具体分子机制;通过干预miR-25-3p的表达,分析并证实其影响肝细胞凋亡、侵袭转移等的机制,为肝脏炎症提供新的分子诊断标志,并为开发基于miRNA的新型抗HBV免疫药物提供依据。
期刊论文列表
专著列表
科研奖励列表
会议论文列表
专利列表
Exosomes secreted by chronic hepatitis B patients with PNALT and liver inflammation grade ≥ A2 promoted the progression of liver cancer by transferring miR-25-3p to inhibit the co-expression of TCF21 and HHIP
PNALT且肝脏炎症等级≥A2级的慢性乙型肝炎患者分泌的外泌体通过转染miR-25-3p抑制TCF21和HHIP的共表达促进肝癌进展
DOI:10.1111/cpr.12833
发表时间:2020-06-11
期刊:CELL PROLIFERATION
影响因子:8.5
作者:Ouyang, Yi;Tang, Yujing;Fu, Xiaoyu
通讯作者:Fu, Xiaoyu
Exosomal microRNA-25 released from cancer cells targets SIK1 to promote hepatocellular carcinoma tumorigenesis
癌细胞释放的外泌体microRNA-25靶向SIK1促进肝细胞癌肿瘤发生
DOI:10.1016/j.dld.2021.07.017
发表时间:2022-07-01
期刊:DIGESTIVE AND LIVER DISEASE
影响因子:4.5
作者:Fu, Xiaoyu;Tang, Yujing;Huang, Yan
通讯作者:Huang, Yan
Integrated Analysis of Long Noncoding RNA Expression Profiles in Acute-on-Chronic Liver Failure.
慢性慢性肝衰竭中长非编码 RNA 表达谱的综合分析
DOI:10.1155/2021/5387856
发表时间:2021
期刊:BioMed research international
影响因子:--
作者:Fu X;Cheng D;Ouyang Y;Li Y;Li R;Peng S;Fu L
通讯作者:Fu L
抗CD4自身抗体介导CD4+T细胞凋亡在艾滋病免疫无应答患者中的机制及治疗作用
- 批准号:--
- 项目类别:面上项目
- 资助金额:54万元
- 批准年份:2021
- 负责人:符小玉
- 依托单位:
国内基金
海外基金
