用模式动物大鼠肝再生和对照与假手术10个时间点的样品经高通量测序，得到microRNAs的动态变化表达谱并鉴定，有78万多条小分子RNA，有注释的约有9600条，与已知序列比对，共有425条可以在库中723条microRNAs找到对应并一致。筛选出11条microRNAs，发现了特异表达差别最明显的两条:miR-182和miR-142，经分析miR-182上调作用促进细胞增殖及分化，而miR-142上调作用抑制细胞增殖及分化并促进细胞凋亡，两者分析结果均与MTT数据一致。合成microRNAs模拟物或抑制物，使microRNAs上调或下调，检测基因表达水平。本项目拟研究miR-182和miR-142对大鼠肝再生的重要调控功能，建立一整套完整的实验体系来研究模式生物microRNAs与肝再生的关系。miR-142可使细胞凋亡，有望鉴定microRNAs类似抑癌基因的功能，筛到治疗癌症的靶标。

The samples in animal models of liver regeneration in rats and their controls were compared with sham operation in 10 time points by high throughput sequencing,we got the dynamic changes of microRNAs expression profiling and identified that there are more than 780 thousands small molecule RNAs in which about 9600 are annotated. Compared with the known sequences, a total of 425 microRNAs can be found consistent with 723 microRNAs in the corresponding database. Then, we screened of 11 microRNAs and found that miR-182 and miR-142 are most obvious difference in specific expression.Baseon our analysis, we found tha the upregulation of miR-182 promotes cell proliferation and differentiation while the upregulation of miR-142 inhibits cell proliferation and differentiation and promote cell apoptosis.This is really consistent with MTT data. Chemically synthesized microRNAs mimics will be performed to detect increase levels of microRNAs expression; as well as chemically synthesized microRNAs inhibitor will be performed to detect decrease levels of microRNAs expression in liver cells. The project is expected to establish a completed set of experimental system to study the relationship between model organisms microRNAs and liver regeneration by studying the important regulatory function of miR-182 and miR-142 on liver regeneration in rats. Because the miR-142 can promote the cell apoptosis, microRNAs is expected to find similar tumor suppressor gene function which may be able to treat cancer.

项目背景：肝脏是机体重要的器官，有代谢、合成、分泌、储存、转运、凝血、解毒和免疫等功能。由于多因会导致肝病，如肝肿瘤、各种肝炎、脂肪肝、酒精肝、肝硬化、急性肝衰竭等。miR-182 在肝切除及肝再生（MAPK细胞增殖信号通路)中参与调控功能并很关键，相关研究未见报道。 研究内容：1.大鼠肝再生相关miR-182 的调控功能验证，合成miR-182 模拟物和miR-182 抑制物可上调或下调和抑制其表达水平， miR-182在体外过表达后促进细胞生长增殖。2.在项目原有研究内容基础上，运用蛋白质+细胞因子芯片，对大鼠肝再生过程中microRNAs（miRNAs）与蛋白质、细胞因子间调控关系进行研究，筛选miRNAs相关的蛋白质和细胞因子，从中选出miR-182到底与哪些蛋白质和细胞因子相关。3.采用生物素双抗体夹心酶联免疫吸附法（ELISA）测定样品中Rela、Casp3、MAP2K1、FGF9、TGFBR、PPP3R1、MAPK9、 MAP3K12、TNFR2、MRAS、BCL2、MEF2C的表达水平。重要结果：1.经高通量测序，得到miRNAs的动态变化表达谱并鉴定，发现了特异表达差别最明显的两条:miR-182和miR-142，进一步研究miR-182促进细胞增殖及分化的调控功能，筛选出一些基因和酶以及蛋白质。从中选出一个caspase-3做功能研究，已投稿一篇英文论文（见附件1）。2．大鼠肝再生中miRNAs与蛋白质、细胞因子间调控关系的研究：运用芯片，筛选miRNAs相关的蛋白质和细胞因子，发现了后两者有一些与miR-182相关。3．ELISA结果：颜色的深浅与大鼠的Rela、Casp3、MAP2K1、FGF9、TGFBR、PPP3R1、MAPK9、 MAP3K12、TNFR2、MRAS、BCL2、MEF2C相关。关键数据：（见正文及附件1）1.英文论文中的数据。2. 经miRNAs与蛋白质、细胞因子间调控关系的研究，分析与挑选出：与miR-182相关的蛋白质+细胞因子芯片结果（【注】1.）。3. ELISA测定样品结果。科学意义：MiRNAs 与肝细胞分化、增殖与凋亡紧密相关。对其调控机制的研究，将有助于发现其调控肝病及治疗的新靶点，具有对肝癌病灶切除后的患者以及献肝者的治疗、康复的临床价值和重要意义。