电针辅助全身麻醉对大鼠心肌缺血再灌注损伤的保护作用及其机制
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河北省科技计划项目(162777196)


Effects and mechanism of electroacupuncture assistant general anesthesia on myocardial reperfusion injury in rats
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    摘要:

    目的 探讨电针辅助全身麻醉对大鼠心肌缺血再灌注损伤的保护作用及其机制。方法 20只9周SPF雄性SD大鼠随机分为对照组和干预组,每组10只。建立大鼠心肌缺血再灌注模型,两组大鼠手术过程均采用七氟烷诱导维持麻醉。干预组大鼠模型在诱导麻醉前30 min,使用电针对大鼠穴位百会、合谷、内关、足三里进行电刺激并维持至手术结束。分别于手术前和手术结束12 h后取各组大鼠外周血;检测外周血中内源性大麻素花生四烯酸乙醇胺(AEA)、促肾上腺皮质激素释放因子(CRF)、肌酸激酶MB型同工酶(CK-MB)、乳酸脱氢酶(LDH)浓度,心肌肌钙蛋白I(cTnI)以及炎症因子白细胞介素1b(IL-1b)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)水平变化。于手术后12 h取大鼠心脏组织,检测大鼠心脏组织缺血区IL-1b、IL-6、TNF-α mRNA表达水平,中性粒细胞浸润,心肌细胞凋亡及半胱氨酸天冬氨酸蛋白酶-3(Caspase 3)活化情况。结果 与手术前比较,对照组和干预组大鼠血浆CK-MB、cTnI、LDH、IL-1b、IL-6、TNF-α水平均显著升高,而干预组大鼠术后血浆CK-MB、cTnI、LDH、IL-1b、IL-6、TNF-α水平显著低于对照组(P<0.05);对照组大鼠血浆中AEA、CRF含量手术前后没有明显变化(P>0.05),而干预组大鼠术后血浆AEA、CRF水平相比于自身手术前及对照组术后显著升高(P<0.05);Realtime PCR显示干预组大鼠心脏组织IL-1b、IL-6、TNF-α表达水平显著低于对照组(P<0.05);流式细胞术和TUNEL染色分析显示干预组大鼠心肌缺血再灌注区中性粒细胞浸润数量及凋亡细胞数量相比于对照组显著减少(P<0.05);Western blot显示干预组活化caspase 3水平相比于对照组显著降低(P<0.05)。结论 电针辅助全身麻醉能够显著降低大鼠缺血再灌注引发的心肌损伤及炎症反应。

    Abstract:

    Objective To explore the effects of electroacupuncture assistant general anesthesia on myocardial reperfusion injury in rats.Methods Rat myocardial reperfusion mode was established, and the rats were randomly separated into control group and intervention group. The rats in two groups received general anesthesia. Besides, rats in intervention group were subjected to electroacupuncture at Baihui (DU20), Hegu (LI4), Neiguan (PC6), Zusanli (ST36) from 30 min before anesthesia induction to the end of operation. The blood of rats before surgery and 12h post-surgery was collected. The arachidonoylet-hanolamine (AEA), corticotrophin-releasing factor (CRF), CK-MB, cTnI, IL-1b, IL-6 and TNF-α in blood were measured by liquid chromatography-mass spectrometry technique or ELISA. The hearts of rats were harvest 12 h post-surgery, and the mRNA expressions of IL-1b, IL-6, TNF-α in hearts were measured by real-time PCR. Flow cytometry, TUNAL assay and western blot were used to evaluate the neutrophil counts, myocardial apoptosis and caspase 3 activation in heart tissue. Results The plasma AEA and CRF in rats of control group had no significant change before or after surgery, whereas the AEA and CRF in rats of intervention group increased significantly compared with before surgery and control group after surgery. Compared with before surgery, the CK-MB, cTnI, LDH, IL-1b, IL-6 and TNF-α increased significantly in two group, while the CK-MB, cTnI, LDH, IL-1b, IL-6 and TNF-α in intervention group post-surgery were lower than control group post-surgery. Flow cytometry and TUNEL assay showed that the infiltrated neutrophils, apoptotic cells and activated caspase 3 in rat heart tissue of intervention group decreased significantly compared with control group. Conclusion Electroacupuncture assistant general anesthesia inhibits the myocardial injury and inflammation response after myocardial ischemia reperfusion in rats.

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  • 在线发布日期: 2022-03-23
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