吴 宾,张 亮,邓龙翔,尤 菲,樊苗苗,李聪叶,袁 铭.双氢杨梅素对缺氧诱导的心肌细胞炎症反应和凋亡的影响及机制[J].,2017,17(34):6632-6636 |
双氢杨梅素对缺氧诱导的心肌细胞炎症反应和凋亡的影响及机制 |
Effects of Dihydromyricetin on Hypoxia-induced Inflammatory Response and Apoptosis in Cardiomyocytes and Underlying Mechanism |
投稿时间:2017-06-14 修订日期:2017-07-10 |
DOI:10.13241/j.cnki.pmb.2017.34.006 |
中文关键词: 双氢杨梅素素 炎症反应 凋亡 CD40 miRNA-145-5p |
英文关键词: Dihydromyricetin Inflammatory response Apoptosis CD40 miRNA-145-5p |
基金项目:陕西省科技攻关项目(S2016YFSF0666) |
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中文摘要: |
摘要 目的:探讨双氢杨梅素对缺氧诱导的心肌细胞炎症和凋亡的影响及其可能调控机制。方法:将心肌细胞(H9C2细胞系)缺氧处理建立细胞模型,随机分为4组:对照组、对照+双氢杨梅素组、缺氧组、缺氧+双氢杨梅素组。对照+双氢杨梅素组和缺氧+双氢杨梅素组分别加入5 μmol/L的双氢杨梅素孵育24 h,对照组和缺氧组分别加入同等体积的DMSO孵育24 h。实时PCR检测心肌细胞CD40、IL-1β、IL-6、TNF-α、miRNA-145-5p的mRNA水平;ELISA检测IL-1β、IL-6和TNF-α的浓度;MTT实验检测心肌细胞生存率;Tunel法检测心肌细胞凋亡;Western blot检测心肌细胞Cleaved Caspase3的表达。结果:与对照组和对照+双氢杨梅素组相比,缺氧组CD40、IL-1β、IL-6、TNF-α的mRNA水平升高(P<0.05),IL-1β、IL-6、TNF-α浓度显著升高(P<0.05),miRNA-145-5p的mRNA水平显著降低(P<0.05),心肌细胞生存率显著降低(P<0.05),心肌细胞凋亡显著增加(P<0.05),Cleaved Caspase3的蛋白表达显著增加;与缺氧组相比,缺氧+双氢杨梅素组CD40、IL-1β、IL-6、TNF-α的mRNA水平显著降低(P<0.05),IL-1β、IL-6、TNF-α浓度显著降低(P<0.05),miRNA-145-5p的mRNA水平显著增加(P<0.05),心肌细胞生存率显著增加(P<0.05),心肌细胞凋亡显著降低(P<0.05),Cleaved Caspase3的蛋白表达显著降低。结论:双氢杨梅素可抑制缺氧诱导的心肌细胞炎症和凋亡,其机制可能与双氢杨梅素上调miRNA-145-5p进一步抑制CD40介导的炎症反应和凋亡有关。 |
英文摘要: |
ABSTRACT Objective: To explore the effects and mechanism of dihydromyricetin on hypoxia-induced inflammatory response and apoptosis in cardiomyocytes. Methods: H9C2 cells were randomly divided to four groups: control group, control + dihydromyricetin group, hypoxia group, hypoxia + dihydromyricetin group. Control + dihydromyricetin group and hypoxia + dihydromyricetin group were treated with dihydromyricetin (5 μmol/L), while control group and hypoxia group were treated with DMSO in the absence or presence of hypoxia for 24 h. The mRNA levels of CD40, IL-1β, IL-6, TNF-α and miRNA-145-5p were evaluated by real-time PCR. The concentrations of IL-1β, IL-6 and TNF-α were measured by ELISA assay. The MTT assay was used to analyze the living cells. Apoptotic index of cardiomyocytes was examined by Tunel. Western blot was used to determine the expression of Cleaved Caspase3. Results: Compared with control group and control + dihydromyricetin group, the mRNA levels of CD40, IL-1β, IL-6 and TNF-α and the concentrations of IL-1β, IL-6 and TNF-α increased significantly (P<0.05), with attenuated miRNA-145-5p mRNA and cell viability (P<0.05), as well as increased apoptotic index (P<0.05), and the expressions of Cleaved Caspase3 were upregulated (P<0.05) in hypoxia group. Compared with those in hypoxia group, inflammatory cytokines reduced significantly (P<0.05) and miRNA-145-5p mRNA increased significantly (P<0.05), with restored cell viability (P<0.05), as well as decreased apoptotic index (P<0.05), and there was a reduction of level of Cleaved Caspase3 (P<0.05) by the treatment with dihydromyricetin in hypoxia + dihydromyricetin group. Conclusion: Dihydromyricetin can significantly decrease hypoxia-induced inflammatory response and apoptosis in cardiomyocytes, possibly by upregulating miRNA-145-5p and further inhibiting CD40-mediated inflammatory response and apoptosis. |
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