首页|香豆素通过调控组蛋白去甲基化酶3和骨髓分化蛋白2改善脓毒症相关AKI的机制研究

香豆素通过调控组蛋白去甲基化酶3和骨髓分化蛋白2改善脓毒症相关AKI的机制研究

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目的 探讨香豆素(Sco)通过调控组蛋白去甲基化酶3(JMJD3)和骨髓分化蛋白2(MD-2)改善脓毒症相关急性肾损伤(AKI)的作用机制.方法 24只C57BL6小鼠根据随机数字表法分为对照组、脂多糖(LPS)组、LPS+40 mg/kg Sco组和LPS+80 mg/kg Sco组.除对照组外,其余3组小鼠腹腔注射LPS 10 mg/kg建立脓毒症相关AKI模型,LPS+40 mg/kg Sco组和LPS+80 mg/kg Sco组小鼠在造模成功后分别腹腔注射Sco 40 mg/kg和80 mg/kg,对照组小鼠腹腔注射0.9%氯化钠溶液2 mL/kg.采用HE染色和原位末端转移酶标记法染色检测小鼠肾脏组织损伤程度和细胞凋亡程度,ELISA法检测小鼠血清血肌酐(Scr)、血尿素氮(BUN)水平,Western blot法检测小鼠肾脏组织中JMJD3和MD-2蛋白表达水平.使用LPS(1 μg/mL)处理人肾小管上皮细胞系HK-2 24 h,建立脓毒症细胞模型,随机分为对照组、LPS组、LPS+30 μmol/L Sco组、LPS+60 μmol/L Sco组、LPS+60 μmol/L Sco+过表达对照组和LPS+60 μmol/L Sco+过表达JMJD3组.采用流式细胞术检测细胞凋亡情况,Western blot法检测细胞中JMJD3和MD-2蛋白表达水平.结果 与对照组小鼠相比,LPS组小鼠肾组织出现明显病理变化,部分肾小管变形,肾小管上皮细胞水肿,肾间质周围炎症细胞浸润,凋亡信号(棕褐色)增强,JMJD3和MD-2蛋白表达水平升高,血清中Scr、BUN水平均升高.而与LPS组相比,LPS+40 mg/kg Sco组和LPS+80 mg/kg Sco组小鼠肾脏组织以上病理变化明显改善,JMJD3和MD-2蛋白表达水平均降低,血清中Scr、BUN水平也均降低,并且LPS+80 mg/kg Sco组变化更明显.与对照组细胞相比,LPS组细胞凋亡水平升高,JMJD3和MD-2蛋白表达水平升高.而相较于LPS组,使用Sco处理后以上变化趋势发生逆转,且Sco浓度越高,趋势变化越显著.此外,过表达JMJD3后细胞凋亡水平升高,JM-JD3和MD-2蛋白表达水平均升高.结论 Sco通过抑制肾小管上皮细胞JMJD3和MD-2表达,减少细胞凋亡,从而改善脓毒症相关AKI.
Scoparone alleviates sepsis-related acute kidney injury mediated by jumonji domain-containing protein 3 and myeloid differ-entiation protein-2
Objective To investigate the mechanism of scoparone(Sco)alleviating sepsis-related acute kidney injury(AKI).Methods Twenty-four C57BL6 mice were randomly divided into control group,LPS group,low-Sco group and high-Sco group with 6 mice in each group.The sepsis-related AKI was induced by intraperitoneal injection of LPS 10 mg/kg in mice of last 3 groups.Mice in low-and high-Sco groups were intraperitoneally injected 40 mg/kg and 80 mg/kg Sco group after successful modeling,respectively;while mice in the control group were intraperitoneally injected with 0.9%sodium chloride solution(2 mL/kg).HE staining and TUNEL staining were used to detect the degree of renal tissue injury and apoptosis.Serum creatinine(Scr)and blood urea nitrogen(BUN)levels were detected by ELISA.The expression levels of JMJD3 and MD-2 protein in renal tissue were detected by Western blot.Human renal tubular epithelial cells HK-2 were treated with LPS(1 μg/mL)for 24 h to establish a sepsis cell model.They were randomly divided into control group,LPS group,LPS+30 μmol/L Sco group,LPS+60 μmol/L Sco group,LPS+60 μmol/L Sco+overexpression control group and LPS+60 μmol/L Sco+JMJD3 overexpression group.Apoptosis was detected by flow cytometry,and the expression levels of JMJD3 and MD-2 protein were detected by Western blot.Results Compared with the control group,the renal tissue of mice in LPS group showed obvious pathological changes,including partial renal tubule deformation,edema of renal tubule epithelial cells,infiltration of perirenal interstitial inflammatory cells,enhanced apoptosis signal(brown),increased expression of JMJD3 and MD-2 protein,and increased serum Scr and BUN levels.Compared with LPS group,the above pathological changes in kidney tissues of mice in LPS+40 mg/kg Sco group and LPS+80 mg/kg Sco group were significantly improved,the expression levels of JMJD3 and MD-2 protein were decreased,and the serum levels of Scr and BUN were also decreased.The change of LPS+80 mg/kg Sco group was more obvious.Compared with the control group,the apoptosis level and the expression level of JMJD3 and MD-2 protein were increased in LPS group.Compared with LPS group,the above change trend was reversed after Sco treatment,and the higher the Sco concentration was,the more significant the change was.In addition,after overexpression of JMJD3,the apoptosis level and the expression levels of JMJD3 and MD-2 protein were significantly increased.Conclusion Sco can alleviate the sepsis-related AKI by inhibiting the expression of JMJD3 and MD-2 in renal tubular epithelial cells,and reducing cell apoptosis.

SepsisAcute kidney injuryScoparoneJumonji domain-containing protein 3Myeloid differentiation protein-2

金光军、赵金凯、徐步海、张建成、潘旭鸣、季明霞

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310005 杭州,浙江中医药大学附属第二医院医务部

浙江中医药大学第二临床医学院

310005 杭州,浙江中医药大学附属第二医院全科医学科

310005 杭州,浙江中医药大学附属第二医院急诊医学科

义乌市中心医院重症医学科

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脓毒症 急性肾损伤 香豆素 蛋白去甲基化酶3 骨髓分化蛋白2

浙江省中医药科技计划

2022ZA079

2024

浙江医学
浙江省医学会

浙江医学

CSTPCD
影响因子:0.428
ISSN:1006-2785
年,卷(期):2024.46(7)
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