Osteonecrosis is a harmful musculoskeletal disease. We aim to detect the effects of icariin (ICA) in MC3T3‐E1 cell. MC3T3‐E1 cell was pretreated with ICA and was subjected to hypoxia stimuli. The tumor‐associated long noncoding RNA expressed on chromosome 2 (TALNEC2) overexpression or silencing vectors (pTALNEC2 or si‐TALNEC2) was utilized for MC3T3‐E1 cell transfection. Viability and apoptosis rate were individually tested by cell counting kit‐8 and Annexin V‐fluorescein isothiocyanate/propidium iodide kit untied with flow cytometry. The alkaline phosphatase activity (ALP) activity was tested through ALP assay. The quantitative reverse transcription PCR or Western blot was performed for elements detection at the RNA or protein level. Hypoxia treatment induced viability inhibition and CyclinD1 reduction, but elevation of p53 and p16. It also promoted apoptosis by increasing apoptotic cells, Bax, and cleaved‐poly ADP‐ribose polymerase but decreasing Bcl‐2. Also, hypoxia stimuli restrained ALP activity, and osteopontin, osteocalcin, and Runt‐related transcription factor 2 expression. Those effects caused by hypoxia stimuli were all reversed by ICA. TALNEC2 was downregulated by ICA, whose impacts were subsequently abolished by pTALNEC2. Silencing TALNEC2 displayed similar effects with ICA. But the apoptosis was not affected by si‐TALNEC2. ICA blocked ste20‐related proline/alanine‐rich kinase/c‐Jun N‐terminal kinase (SPAK/JNK) but triggered phosphatidylinositol 3‐kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) pathway in MC3T3‐E1 cell by suppressing TALNEC2. ICA relieved hypoxia‐stimulated damage by restraining TALNEC2 through blocking SPAK/JNK and triggering PI3K/AKT/mTOR in the MC3T3‐E1 cell.

中文翻译：

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鹰嘴豆素通过下调TALNEC2减轻MC3T3-E1细胞缺氧诱导的损伤。

骨坏死是一种有害的肌肉骨骼疾病。我们旨在检测icariin（ICA）对MC3T3-E1细胞的作用。MC3T3-E1细胞经过ICA预处理，并受到缺氧刺激。在2号染色体（TALNEC2）过表达或沉默载体（pTALNEC2或si-TALNEC2）上表达的与肿瘤相关的长非编码RNA被用于MC3T3-E1细胞转染。活力和凋亡率通过细胞计数试剂盒-8和膜联蛋白V-异硫氰酸荧光素/碘化丙啶试剂盒单独检测，未与流式细胞仪捆绑。通过ALP测定法测试碱性磷酸酶活性（ALP）活性。进行定量逆转录PCR或蛋白质印迹以在RNA或蛋白质水平上进行元素检测。缺氧处理可诱导活力抑制和CyclinD1降低，但p53和p16升高。它还通过增加凋亡细胞，Bax和裂解的ADP-核糖聚合酶，但降低Bcl-2来促进细胞凋亡。此外，缺氧刺激会抑制ALP活性，并抑制骨桥蛋白，骨钙蛋白和Runt相关转录因子2的表达。由缺氧刺激引起的那些作用全部被ICA逆转。ICA下调了TALNEC2的功能，随后pTALNEC2取消了其影响。使TALNEC2静音对ICA表现出相似的效果。但是细胞凋亡不受si-TALNEC2的影响。ICA阻断了MC3T3‐中与Ste20相关的脯氨酸/富含丙氨酸的激酶/ c‐Jun N末端激酶（SPAK / JNK），但触发了磷脂酰肌醇3-激酶/蛋白激酶B /雷帕霉素的哺乳动物靶点（PI3K / AKT / mTOR）通过抑制TALNEC2来激活E1细胞。