Background Myocardial ischemia-reperfusion injury is an important clinical problem.Hypoxia-reperfusion(HR)triggers a series of complex cellular reactions and pathological changes.In recent years,the role of N-6-methyl...Background Myocardial ischemia-reperfusion injury is an important clinical problem.Hypoxia-reperfusion(HR)triggers a series of complex cellular reactions and pathological changes.In recent years,the role of N-6-methyladenosine(m6A)modification in gene expression regulation has received extensive attention.New evidence suggests that m6A modifications may play a key role in the regulation of cellular responses to HR-damaged cardiomyocytes.Inflammation is an important part of the pathogenesis of myocardial ischemia-reperfusion injury.In the HR response,inflammatory factors such as interleukin(IL),tumor necrosis factor(TNF-α)and other cytokines are released and activated,which can trigger a series of intracellular signaling pathways leading to myocardial cell damage,apoptosis,and inflammation.In the context of cardiomyocyte H/R injury,it is important to study the relationship between m6A modification and the regulation of inflammatory cytokines expression.It is of great significance to fully understand the underlying mechanism of myocardial ischemia-reperfusion injury and develop new therapeutic strategies.Methods The hypoxic-reoxygenation model of H9C2 cells was established,and the overexpression and knockout model of fat mass and obesity associated gene(FTO)was established.The cells were divided into 6 groups:control group(CG),hypoxic-reperfusion group(HRG),FTO overexpression group(OG),FTO overexpression hypoxic-reperfusion group(OHG),FTO knockout group(KG)and FTO knockout hypoxic-reperfusion group(KHG).The changes of M6A were detected by colorimetry,FTO expression was detected by Western Blot,and the changes of FTO and phosphorylated nuclear factor kappa B(P-NF-κB)were observed by immunofluorescence.IL-6 was detected by enzymelinked immunosorbent assay(ELISA).Results The level of m6A in HRG was significantly higher than that in CG(P<0.05).The same pattern was observed in OG and KG(P<0.05).The level of m6A in KHG were higher than that in HRG,while lower in OHG than that in HRG(P<0.05).The expression of FTO decreased during hypoxia and reoxygenation(P<0.05).The expression of FTO in KHG was lower than that in KG(P<0.05).There was no significant difference in FTO expression between OG and OHG(P>0.05).Compared with HRG,FTO in OHG was decreased(P<0.01).Compared with HRG,FTO in KHG was decreased(P<0.01).The changes of P-NF-κB and IL-6 were consistent before and after hypoxia reoxygenation.The expression of these two inflammatory factors in HRG was significantly higher than that in CG(P<0.05).OHG compared with HRG,the two groups of inflammatory factors decreased;KHG was significantly higher than HRG(P<0.05).Conclusions Overexpression of FTO can significantly inhibit the increase of m6A,reduce inflammatory factors,and protect damaged cells during hypoxia and re-oxidation processes.[S Chin J Cardiol 2024;25(3):169-178]展开更多
本研究在明确姜黄素有效治疗肥胖小鼠溃疡性结肠炎的基础上,评价其对结肠炎肥胖小鼠记忆性滤泡性T细胞(memory follicular T cells,mTf)的调控作用。经江西中医药大学实验动物伦理委员会审议同意并批准后,将40只瘦素突变型(ob/ob)雄性...本研究在明确姜黄素有效治疗肥胖小鼠溃疡性结肠炎的基础上,评价其对结肠炎肥胖小鼠记忆性滤泡性T细胞(memory follicular T cells,mTf)的调控作用。经江西中医药大学实验动物伦理委员会审议同意并批准后,将40只瘦素突变型(ob/ob)雄性小鼠随机分为对照组、对照+姜黄素组、模型组和模型+姜黄素组,每组10只。采用葡聚糖硫酸钠(dextran sodium sulfate,DSS)诱导模型组和模型+姜黄素组小鼠建立慢性溃疡性结肠炎模型后,对照+姜黄素组和模型+姜黄素组小鼠灌胃给予姜黄素(200 mg·kg^(-1)·d^(-1))。麻醉处死小鼠,观察各组小鼠结肠质量、结肠质量指数、结肠长度等情况;HE染色后进行病理损伤评分;流式细胞术检测小鼠脾脏记忆性滤泡性辅助性T细胞(memory follicular helper T cells,mTfh)和记忆性滤泡性调节性T细胞(memory follicular regulatory T cells,mTfr)细胞亚群水平;酶联免疫吸附法检测结肠组织中白介素-10(interleukin-10,IL-10)和白介素-17A(interleukin-17A,IL-17A)的表达量。结果表明,姜黄素可明显增加结肠炎肥胖小鼠的体重和结肠长度,降低结肠重量、结肠质量指数和病理评分(P<0.05);并且姜黄素可显著降低中枢型记忆性滤泡性T细胞(central memory follicular T cells,cmTf)、mTfh1、mTfh17细胞水平和促炎细胞因子IL-17A含量(P<0.01),升高效应型记忆性滤泡性T细胞(effector memory follicular T cells,emTf)、mTfr细胞水平及抑炎细胞因子IL-10含量(P<0.05,P<0.01)。因此,姜黄素治疗肥胖小鼠结肠炎,可能是通过调控mTf细胞亚群平衡来实现的。展开更多
基金supported by grants from Shenzhen Science and Technology Project(No.JCYJ20220531091817039)。
文摘Background Myocardial ischemia-reperfusion injury is an important clinical problem.Hypoxia-reperfusion(HR)triggers a series of complex cellular reactions and pathological changes.In recent years,the role of N-6-methyladenosine(m6A)modification in gene expression regulation has received extensive attention.New evidence suggests that m6A modifications may play a key role in the regulation of cellular responses to HR-damaged cardiomyocytes.Inflammation is an important part of the pathogenesis of myocardial ischemia-reperfusion injury.In the HR response,inflammatory factors such as interleukin(IL),tumor necrosis factor(TNF-α)and other cytokines are released and activated,which can trigger a series of intracellular signaling pathways leading to myocardial cell damage,apoptosis,and inflammation.In the context of cardiomyocyte H/R injury,it is important to study the relationship between m6A modification and the regulation of inflammatory cytokines expression.It is of great significance to fully understand the underlying mechanism of myocardial ischemia-reperfusion injury and develop new therapeutic strategies.Methods The hypoxic-reoxygenation model of H9C2 cells was established,and the overexpression and knockout model of fat mass and obesity associated gene(FTO)was established.The cells were divided into 6 groups:control group(CG),hypoxic-reperfusion group(HRG),FTO overexpression group(OG),FTO overexpression hypoxic-reperfusion group(OHG),FTO knockout group(KG)and FTO knockout hypoxic-reperfusion group(KHG).The changes of M6A were detected by colorimetry,FTO expression was detected by Western Blot,and the changes of FTO and phosphorylated nuclear factor kappa B(P-NF-κB)were observed by immunofluorescence.IL-6 was detected by enzymelinked immunosorbent assay(ELISA).Results The level of m6A in HRG was significantly higher than that in CG(P<0.05).The same pattern was observed in OG and KG(P<0.05).The level of m6A in KHG were higher than that in HRG,while lower in OHG than that in HRG(P<0.05).The expression of FTO decreased during hypoxia and reoxygenation(P<0.05).The expression of FTO in KHG was lower than that in KG(P<0.05).There was no significant difference in FTO expression between OG and OHG(P>0.05).Compared with HRG,FTO in OHG was decreased(P<0.01).Compared with HRG,FTO in KHG was decreased(P<0.01).The changes of P-NF-κB and IL-6 were consistent before and after hypoxia reoxygenation.The expression of these two inflammatory factors in HRG was significantly higher than that in CG(P<0.05).OHG compared with HRG,the two groups of inflammatory factors decreased;KHG was significantly higher than HRG(P<0.05).Conclusions Overexpression of FTO can significantly inhibit the increase of m6A,reduce inflammatory factors,and protect damaged cells during hypoxia and re-oxidation processes.[S Chin J Cardiol 2024;25(3):169-178]
文摘本研究在明确姜黄素有效治疗肥胖小鼠溃疡性结肠炎的基础上,评价其对结肠炎肥胖小鼠记忆性滤泡性T细胞(memory follicular T cells,mTf)的调控作用。经江西中医药大学实验动物伦理委员会审议同意并批准后,将40只瘦素突变型(ob/ob)雄性小鼠随机分为对照组、对照+姜黄素组、模型组和模型+姜黄素组,每组10只。采用葡聚糖硫酸钠(dextran sodium sulfate,DSS)诱导模型组和模型+姜黄素组小鼠建立慢性溃疡性结肠炎模型后,对照+姜黄素组和模型+姜黄素组小鼠灌胃给予姜黄素(200 mg·kg^(-1)·d^(-1))。麻醉处死小鼠,观察各组小鼠结肠质量、结肠质量指数、结肠长度等情况;HE染色后进行病理损伤评分;流式细胞术检测小鼠脾脏记忆性滤泡性辅助性T细胞(memory follicular helper T cells,mTfh)和记忆性滤泡性调节性T细胞(memory follicular regulatory T cells,mTfr)细胞亚群水平;酶联免疫吸附法检测结肠组织中白介素-10(interleukin-10,IL-10)和白介素-17A(interleukin-17A,IL-17A)的表达量。结果表明,姜黄素可明显增加结肠炎肥胖小鼠的体重和结肠长度,降低结肠重量、结肠质量指数和病理评分(P<0.05);并且姜黄素可显著降低中枢型记忆性滤泡性T细胞(central memory follicular T cells,cmTf)、mTfh1、mTfh17细胞水平和促炎细胞因子IL-17A含量(P<0.01),升高效应型记忆性滤泡性T细胞(effector memory follicular T cells,emTf)、mTfr细胞水平及抑炎细胞因子IL-10含量(P<0.05,P<0.01)。因此,姜黄素治疗肥胖小鼠结肠炎,可能是通过调控mTf细胞亚群平衡来实现的。
