miRNA studies in in vitro and in vivo activated hepatic stellate cells

AIM: To understand which and how different miRNAs are implicated in the process of hepatic stellate cell (HSC) activation. METHODS: We used microarrays to examine the differential expression of miRNAs during in vitro activation of primary HSCs (pHSCs). The transcriptome changes upon stable transfection of rno-miR-146a into an HSC cell line were studied using cDNA microarrays. Selected differentially regulated miRNAs were investigated by quantitative real-time polymerase chain reaction during in vivo HSC activation. The effect of miRNA mimics and inhibitor on the in vitro activation of pHSCs was also evaluated.RESULTS: We found that 16 miRNAs were upregulated and 26 were downregulated significantly in 10-d in vitro activated pHSCs in comparison to quiescent pHSCs. Overexpression of rno-miR-146a was characterized by marked upregulation of tissue inhibitor of metalloproteinase-3, which is implicated in the regulation of tumor necrosis factor-α activity. Differences in the regulation of selected miRNAs were observed comparing in vitro and in vivo HSC activation. Treatment with miR-26a and 29a mimics, and miR-214 inhibitor during in vitro activation of pHSCs induced significant downregulation of collagen type Ⅰ transcription. CONCLUSION: Our results emphasize the different regulation of miRNAs in in vitro and in vivo activated pHSCs. We also showed that miR-26a, 29a and 214 are involved in the regulation of collagen type I mRNA.

GFAP promoter directs lacZ expression specifically in a rat hepatic stellate cell line

瞄准：GFAP 传统地被认为是为神经神经胶质的一个简历标记(主要星形细胞和 non-myelinating Schwann 房间) 。遗传上，一个 2.2-kb 人 GFAP 倡导者成功地被用来指向星形细胞在试管内和在活体内。更最近， GFAP 也为识别肝的星形细胞(HSC ) 作为几个制造者之一被建立。在这个工程，为指向 HSC 的一样的 2.2-kb 人 GFAP 倡导者的可能的申请被调查。方法：GFAP-lacZ transgene 是进各种各样的房间线(HSC， hepatocyte，和另外的 non-HSC 房间类型) 的 transfected。transgene 表示特性被贝它牛乳糖活动染色的 X 女郎决定。并且 transgene 的应答的海角与典型 pro-fibrotic cytokine TGF-beta1 被测试。内长的 GFAP 基因的表示被即时 RT-PCR 估计，提供为 transgene 表示的一本参考书。结果：结果第一次证明 2.2 kb hGFAP 倡导者不仅能够指导 HSC 特定的表示，而且对已知的 pro-fibrogenic cytokine TGF-beta1 作出回应由在在一个剂量依赖者和时间依赖者举止的规定上面，类似于内长的 GFAP。结论：在结论，这些调查结果为使用 GFAP 倡导者明确地为治疗学的目的操作 HSC 建议了新奇实用程序。