美洲大蠊提取物对脓毒症小鼠...-1α、UCP2表达的影响_李洁.pdf

美洲大蠊提取物对脓毒症小鼠心肌的保护作用及 PGC 1、UCP2 表达的影响李洁1，杜金穗2，唐品清2，吴冬雪2，董辰辰2(1 东南大学附属中大医院，江苏 南京 210009;2 东南大学医学院，江苏 南京210009)摘要 目的探讨美洲大蠊提取物(心脉隆注射液)对脓毒症小鼠心肌损伤及心肌线粒体过氧化物酶体增殖物激活受体 共激活因子 1(PGC 1)、解耦联蛋白 2(UCP2)蛋白表达的影响。方法取8 10 周雄性 C57BL/6J 小鼠30 只，随机分为空白组、脓毒症组、心脉隆注射液组，每组 10 只。空白组和脓毒症组连续 3 d 腹腔注射生理盐水，心脉隆注射液组连续 3 d 腹腔注射心脉隆注射液 200 mg/(kgd)，脓毒症组和心脉隆注射液组第 3 天同时腹腔注射脂多糖 10 mg/kg。干预结束后，应用全自动生化分析仪检测血清乳酸脱氢酶 1(LDH 1)及肌酸磷酸激酶同工酶(CK MB)水平，应用黄嘌呤氧化酶法检测血清超氧化物歧化酶(SOD)活力，应用分光光度法检测血清丙二醛(MDA)水平，HE 染色观察心肌组织病理表现，酶联免疫吸附法检测心肌组织中肿瘤坏死因子 (TNF )含量，Western blot 法检测心肌线粒体中 PGC 1、UCP2 蛋白表达情况。结果与空白组比较，脓毒症组小鼠血清 LDH 1、CK MB、MDA 水平均明显升高(P 均 0 05)，SOD 水平明显降低(P 0 05);与脓毒症组比较，心脉隆注射液组小鼠血清 LDH 1、CK MB、MDA 水平均明显降低(P 均 0 05)，SOD 水平明显增高(P 0 05)。HE 染色显示，空白组小鼠心肌组织结构正常;脓毒症组小鼠心肌细胞排列紊乱，有炎性细胞浸润;心脉隆注射液组小鼠心肌组织中炎性细胞浸润较脓毒症组少。与空白组比较，脓毒症组心肌组织中 TNF 含量明显增高(P 0.05)，心肌线粒体中 PGC 1、UCP2 蛋白表达量均明显降低(P 均 0 05);与脓毒症组比较，心脉隆注射液组心肌组织中 TNF 含量明显降低(P 0 05)，心肌线粒体中 PGC 1、UCP2 蛋白表达量均明显升高(P 均 0 05)。结论美洲大蠊提取物对于脓毒症小鼠心肌具有保护作用，其机制与 PGC 1/UCP2 介导的抗炎、抑制氧化应激作用密切相关。关键词 美洲大蠊提取物;脓毒症;心肌损伤;过氧化物酶体增殖物激活受体 共激活因子 1;解耦联蛋白 2doi:10 3969/j issn 1008 8849 2022 22 003 中图分类号 332 文献标识码 A 文章编号 1008 8849(2022)22 3078 05 作者简介 李洁，女，博士，主任医师，硕士研究生导师，研究方向为中西医结合防治心血管疾病。基金项目 江苏省中医药科技发展计划项目(YB201991)Protective effect of Periplaneta americana extract on myocardium of septic mice and its effect onPGC-1，UCP2 expressionLI Jie1，DU Jinsui2，TANG Pinqing2，WU Dongxue2，DONG Chenchen2(1 Zhongda Hospital Affiliated to Southeast University，Nanjing 210009，Jiangsu，China;2 School of Medicine of South-east University，Nanjing 210009，Jiangsu，China)Abstract:Objective It is to explore the effects of Periplaneta americana extract(Xinmailong injection)on myocardialdamage and expression of myocardial mitochondrial peroxisome proliferator activated receptor coactivator-1(PGC-1)，uncoupling protein 2(UCP2)in septic mice Methods Thirty male C57BL/6J mice aged 8-10 weeks were selected andrandomly divided into blank group，sepsis group and Xinmailong injection group，with 10 mice in each group The blankgroup and sepsis group were intraperitoneally injected with normal saline for 3 consecutive days，the Xinmailong injectiongroup was intraperitoneally injected with Xinmailong injection 200 mg/(kgd)for 3 consecutive days，and the sepsis8703现代中西医结合杂志 Modern Journal of Integrated Traditional Chinese and Western Medicine 2022 Nov，31(22)group and Xinmailong injection group were intraperitoneally injected with lipopolysaccharide 10 mg/kg at the same time onthe third day After the intervention，the levels of serum lactate dehydrogenase-1(LDH-1)and creatine phosphokinaseisoenzyme(CK-MB)were detected by automatic biochemical analyzer，the activity of serum superoxide dismutase(SOD)was detected by xanthine oxidase method，the level of serum malondialdehyde(MDA)was detected by spectrophotometry，the pathological manifestation of myocardial tissue was observed by HE staining，and the content of tumor necrosis factor-(TNF-)in myocardial tissue was detected by enzyme linked immunosorbent assay，the expression of PGC-1，UCP2 pro-tein in myocardial mitochondria was detected by Western blot esults Compared with the blank group，the levels of LDH-1，CK-MB and MDA in serum of mice in sepsis group were significantly increased(all P 0 05)，while the level of SODwas significantly decreased(P 0 05);Compared with the sepsis group，the levels of LDH-1，CK-MB and MDA in serumof mice in Xinmailong injection group were significantly decreased(all P 0 05)，and the level of SOD was significantlyincreased(P 0 05)HE staining showed that the myocardial tissue structure was normal in the blank group;The myo-cardial cells were disorderly arranged with inflammatory cell infiltration in the sepsis group;The inflammatory cell infiltra-tion in myocardial tissue of mice in Xinmailong injection group was less than that in sepsis group Compared with the blankgroup，the content of TNF-in myocardial tissue of sepsis group was significantly increased(P 0 05)，the expression ofPGC-1 and UCP2 protein in myocardial mitochondria were significantly decreased(all P 0 05);Compared with thesepsis group，the content of TNF-in myocardial tissue of sepsis group was significantly decreased(P 0 05)，the ex-pression of PGC-1 and UCP2 protein in myocardial mitochondria were significantly increased(all P 0 05)ConclusionPeriplaneta americana extract could protect myocardium of septic mice，and its mechanism is closely related to PGC-1/UCP2-mediated anti-inflammatory and inhibition of oxidative stressKeywords:Periplaneta americana extract;sepsis;myocardial injury;peroxisome proliferator-activated receptor-co-activator factor-1;decoupling protein 2脓毒症是由感染诱发的机体系统性炎症反应，通过激活免疫系统释放多种炎症因子，免疫失衡的炎症反应会导致全身多系统、多器官功能障碍，是临床重症监护病房危重患者的死亡常见原因1。脓毒症导致的心肌损伤累及心脏收缩和舒张功能，一旦并发休克出现血流动力学异常，通常预后较差，故早期干预脓毒症心肌损伤具有重要的临床意义2。脓毒症心肌损伤的发病机制与机体炎症反应、氧化应激、一氧化氮及能量代谢失衡等因素密切相关。线粒体是合成三磷酸腺苷(ATP)的主要细胞器，其还参与钙储备调控、细胞信息传递以及细胞凋亡调控，对各种损伤极为敏感，是防治脓毒症心肌损伤的主要干预靶点。脓毒症发生时，活性氮和氧自由基的生成增加，线粒体内外钙稳态失衡，线粒体嵴被破坏，引起线粒体发生肿胀、碎裂，损伤线粒体有氧呼吸功能，导致合成 ATP 功能障碍3。目前研究证实，中医药可从线粒体能量代谢、线粒体融合与分裂、线粒体氧化应激等多个方面调控机体五脏阴阳平衡，阐明了中医药以细胞