| 刘亚东,卢小伟.miR-495-3p联合黄氏总皂苷对高糖诱导的成骨细胞增殖、凋亡的影响及其机制研究[J].安徽医药,2021,25(4):834-839. |
| miR-495-3p联合黄氏总皂苷对高糖诱导的成骨细胞增殖、凋亡的影响及其机制研究 |
| EffectofmiR-495-3pcombinedwithtotalsaponinsof Astragalusmembranaceus onproliferation and apoptosis of osteoblasts induced by high glucose and its mechanism |
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| DOI：10.3969/j.issn.1009-6469.2021.04.049 |
| 中文关键词: 微 RNAs 中草药 miR-495-3p 黄芪总皂苷 高血糖症 成骨细胞 增殖 凋亡 |
| 英文关键词: MicroRNAs Drugs, Chinese herbal miR-495-3p Total saponins of Astragalus membranaceus Hyperglycemia Osteoblasts Proliferation Apoptosis |
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| 中文摘要: |
| 目的探讨微小 RNA-495-3p(miR-495-3p)联合黄芪总皂苷(TSA)在高糖诱导的成骨细胞增殖、凋亡中的作用及其机制。方法 2018年 2月至 2019年 4月，将商购的小鼠成骨细胞 MC3T3-E1用随机数字表法分为对照(NC)组，高糖组，高糖 +不同浓度 TSA组，高糖 +miRNA阴性对照(miR-con)组，高糖 +miR-495-3p组，高糖 +TSA+抗 miRNA阴性对照(anti-miR-con)组，糖+TSA+抗 miR-495-3p(anti-miR-495-3p)。噻唑蓝(MTT)法检测细胞增殖，流式细胞仪检测细胞凋亡，实时定量 PCR(qRT高PCR)检测 miR-495-3p表达， Western blotting检测细胞周期蛋白 D1(CyclinD1)、活化的含半胱氨酸的天冬氨酸蛋白水解酶 3(Cleave-caspase-3)蛋白水平。结果与 NC组比较，高糖组 MC3T3-E1细胞存活率［(52.26±5.26)%比(100.26±10.03)%］和 miR495-3p［(0.25±0.03)比(1.00±0.10)］、 CyclinD1表达量明显降低，细胞凋亡率［(35.33±3.53)%比(10.06±1.51)%］和 Cleave-caspase-3蛋白水平显著升高(P<0.05)。与高糖组比较， 0.5 μg/mL、1.0 μg/mL、2.0 μg/mL、5.0 μg/mL、10 μg/mL浓度 TSA显著提高 MC3T3-E1细胞存活率［(63.26±6.02)%、(72.56±7.25)%、(90.46±8.88)%、(100.04±10.01)%、(102.56±10.23)%比(52.26±5.26)%］，2.0 μg/mL TSA显著降低细胞凋亡率［(18.56±1.66)%比(35.33±3.53)%］和 Cleave-caspase-3水平，明显提高 miR-4953p［(0.95±0.09)比(0.25±0.03)］、 CyclinD1表达量(P<0.05)，这与高糖 +miR-495-3p组促进 MC3T3-E1增殖，抑制凋亡的作用相同。低表达 miR-495-3p可以逆转 TSA促进高糖诱导的 MC3T3-E1细胞增殖作用和抑制细胞凋亡作用。结论黄芪总皂苷通过 miR-495-3p促进高糖诱导的成骨细胞增殖，并抑制其凋亡。 |
| 英文摘要: |
| Objective To investigate the effects of microRNA-495-3p (miR-495-3p) combined with total saponins of Astragalus membranaceus (TSA) on the proliferation and apoptosis of osteoblasts induced by high glucose and its mechanism.Methods From February 2018 to April 2019, purchased mouse osteoblasts MC3T3-E1 were randomly assigned into control (NC) group, high glucose group, high-glucose + different concentration TSA group, high glucose +miRNA negative control (miR-con) group, high glucose+miR495-3p group, high glucose+TSA+anti-miRNA negative control (anti-miR-con) group, high glucose+TSA+anti-miR-495-3p (anti-miR495-3p) group. Methyl thiazolyl tetrazolium (MTT) assay was used to detect cell proliferation, flow cytometry was applied to evaluatecell apoptosis, Quantitative real-time PCR (qRT-PCR) was used to determine the expression of miR-495-3p, and Western blotting was employed to analyze the levels of CyclinD1 (CyclinD1) and active cysteine-containing Cleave-caspase-3 protein.Results Compared with NC group, the survival rate [(52.26±5.26)% vs (100.26±10.03)%] of MC3T3-E1 cells and the expressions of miR-495-3p [(0.25± 0.03) vs (1.00±0.10)] and CyclinD1 in high glucose group were evidently decreased, while the apoptotic rate [(35.33±3.53)% vs (10.06± 1.51)%] and the level of Cleave-caspase-3 protein were greatly increased (P < 0.05). Compared with the high glucose group, 0.5 μg/mL, |
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