Screening and Identification of Key Genes for Activation of Islet Stellate Cell.
Screening and Identification of Key Genes for Activation of Islet Stellate Cell.
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胰岛星状细胞激活关键基因的筛选与鉴定
DOI：
10.3389/fendo.2021.695467
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发表时间：
2021
影响因子：
5.2
通讯作者：
Sun Z
中科院分区：
文献类型：
--
作者：
Wang X;Carvalho V;Wang Q;Wang J;Li T;Chen Y;Ni C;Liu L;Yuan Y;Qiu S;Sun Z
背景研究表明，活化的胰岛星状细胞（ISCs）在胰岛纤维化中起着重要作用，并在2型糖尿病的发生发展中起重要作用。然而，负责ISCs激活的关键分子尚未确定。本研究的目的是确定参与糖尿病诱导的ISCs激活的潜在关键基因。方法分离10周龄健康雄性Wistar大鼠和Goto-Kakizaki（GK）大鼠各3只的星状细胞。在相同条件下对每只大鼠的细胞进行原代培养。使用Hiseq 3000平台生成星状细胞的全基因组转录序列。采用实时荧光定量PCR和蛋白质印迹法对GK大鼠、高脂饮食（HFD）大鼠及其对照组的差异表达基因进行验证。结果共获得204条GK间差异表达基因（DEG）。ISCs和Wistar ISCs（W.ISCs）占检测到的35，362个基因的0.58%。在基因本体（GO）和基因和基因组的京都百科全书（KEGG）富集分析之后，通过实时PCR进一步证实这些基因在培养的ISCs中的mRNA水平。然后我们选择Fos、Pdpn、Bad作为糖尿病诱导ISCs活化的潜在关键基因。最后，我们通过免疫印迹法和免疫荧光法证实了GK大鼠、HFD大鼠及其对照组中FOS、podoplanin和Bad的蛋白表达水平。结果显示，与对照组相比，GK、ISC和HFD大鼠FOS的表达水平明显降低，而podoplanin和Bad的表达水平明显升高，这与α-平滑肌肌动蛋白的表达一致。结论GK.ISCs与W. ISCs之间共检出204个DEG。在对GK大鼠和HFD大鼠的潜在关键基因表达进行验证后，Fos、Pdpn和Bad可能是参与糖尿病诱导的ISCs激活的潜在关键基因。
Background It has been demonstrated that activated islet stellate cells (ISCs) play a critical role in islet fibrogenesis and significantly contribute to the progression of type 2 diabetes mellitus. However, the key molecules responsible for ISCs activation have not yet been determined. This study aimed to identify the potential key genes involved in diabetes-induced activation of ISCs. Method Stellate cells were isolated from three 10-week-old healthy male Wistar rats and three Goto-Kakizaki (GK) rats. Cells from each rat were primary cultured under the same condition. A Genome-wide transcriptional sequence of stellate cells was generated using the Hiseq3000 platform. The identified differentially expressed genes were validated using quantitative real-time PCR and western blotting in GK rats, high fat diet (HFD) rats, and their controls. Results A total of 204 differentially expressed genes (DEGs) between GK. ISCs and Wistar ISCs (W.ISCs) were identified, accounting for 0.58% of all the 35,362 genes detected. After the Gene Ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses, the mRNA levels of these genes were further confirmed by real-time PCR in cultured ISCs. We then selected Fos, Pdpn, Bad as the potential key genes for diabetes-induced activation of ISCs. Finally, we confirmed the protein expression levels of FOS, podoplanin, and Bad by western blotting and immunofluorescence in GK rats, HFD rats, and their controls. The results showed that the expression level of FOS was significantly decreased, while podoplanin and Bad were significantly increased in GK.ISCs and HFD rats compared with controls, which were consistent with the expression of α-smooth muscle actin. Conclusions A total of 204 DEGs were found between the GK.ISCs and W.ISCs. After validating the expression of potential key genes from GK rats and HFD rats, Fos, Pdpn, and Bad might be potential key genes involved in diabetes-induced activation of ISCs.
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影响因子：
37.3
作者：
Shindo K;Aishima S;Ohuchida K;Fujiwara K;Fujino M;Mizuuchi Y;Hattori M;Mizumoto K;Tanaka M;Oda Y
通讯作者：
Oda Y
DOI：
10.1186/1755-1536-2-1
发表时间：
2009-01-30
期刊：
Fibrogenesis & tissue repair
影响因子：
--
作者：
Gentilini, Alessandra;Lottini, Benedetta;Brogi, Marco;Caligiuri, Alessandra;Cosmi, Lorenzo;Marra, Fabio;Pinzani, Massimo
通讯作者：
Pinzani, Massimo
影响因子：
4.3
作者：
Li FF;Chen BJ;Li W;Li L;Zha M;Zhou S;Bachem MG;Sun ZL
通讯作者：
Sun ZL
DOI：
10.1016/j.bbrc.2011.09.087
发表时间：
2011-10-22
影响因子：
3.1
作者：
Lee, Esder;Ryu, Gyeong Ryul;Song, Ki-Ho
通讯作者：
Song, Ki-Ho
影响因子：
7.7
作者：
Chiasson, JL;Rabasa-Lhoret, M
通讯作者：
Rabasa-Lhoret, M