摘要
目的观察含血必净注射液的供心保存灌注液对体外膜肺氧合(ECMO)支持下离体空跳猪心的心肌保护作用,并探讨其可能的机制。方法选取健康成年广西巴马小型猪12只,随机分成血必净组和对照组,每组6只。血必净组和对照组均采用ECMO支持猪离体空跳心脏,分别用含50 mL血必净注射液、生理盐水的去白细胞氧合血连续灌注,维持心脏空跳8 h。灌注0 h(T1)、2 h(T2)、4 h(T3)、6 h(T4)、8 h(T5),记录两组心率、灌注压、灌注流量,采用ELISA法检测各时间点灌注液炎症相关指标白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)水平。使用全自动生化分析仪检测两组全身肝素化后且灌注前(T0)及T1~T5时间点灌注液心肌损伤标志物cTnT(cTnT)、肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)水平。透射电镜下观察两组T5时间点的心肌组织超微结构。结果两组T1~T5时间点心率、灌注压、灌注流量比较差异均无统计学意义(P均>0.05)。与同组T1时间点比较,对照组T3~T5时间点灌注液IL-1β、IL-6、TNF-α、cTnT、CK、CK-MB、LDH水平均升高,血必净组T4~T5时间点灌注液cTnT、CK、CK-MB、LDH水平均升高(P均<0.05);血必净组T3~T5时间点灌注液IL-1β、IL-6、TNF-α、cTnT水平均低于对照组,T2~T5时间点灌注液CK、CK-MB、LDH水平均低于对照组(P均<0.05)。心肌组织超微结构:对照组心肌组织中重度退行性变、脂肪变,细胞内基质大面积溶解,肌纤维结构排列紊乱、大量断裂,线粒体大多肿胀严重、大小不均,细胞膜溶解、破损;与对照组比较,血必净组细胞结构损伤相对较轻,心肌细胞轻度肥大,线粒体轻度肿胀、大小均一,肌纤维粗细均匀、排列整齐。结论含血必净注射液的供心保存灌注液对ECMO支持下离体空跳猪心具有心肌保护作用,其机制可能与抑制炎症反应、减轻心肌细胞线粒体损伤有关。
Objective To observe the myocardial protective effect of donor heart preservation perfusate containing Xuebijing injection on the isolated beating pig heart preserved by extracorporeal membrane oxygenation(ECMO),and to explore its possible mechanism.Methods Twelve healthy adult Guangxi Bama miniature pigs were randomly divided in⁃to the control group and Xuebijing group,with 6 pigs in each group.The ECMO-preserved beating heart models were estab⁃lished in the control group and Xuebijing group.The models were continuously perfused with leukocyte-depleted oxygenat⁃ed blood containing 50 mL Xuebijing injection and normal saline,respectively,to maintain the beating heart for 8 h.Heart rate,perfusion pressure and perfusion flow were recorded at 0(T1),2(T2),4(T3),6(T4)and 8 h(T5)after perfusion.The levels of interleukin-1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were detected by ELISA.The myocardial injury markers troponin I(cTnI),creatine kinase(CK),creatine kinase isoenzyme(CK-MB)and lactate dehydrogenase(LDH)were detected by automatic biochemical analyzer after heparinization and before perfu⁃sion(T0)and at T1-T5 time points.The ultrastructure of myocardial tissue in the two groups at T5 was observed under transmission electron microscope.Results There were no significant differences in heart rate,perfusion pressure or per⁃fusion flow between the two groups at T1-T5(all P>0.05).The levels of serum IL-1β,IL-6,TNF-αand perfusion fluid cTnT,CK,CK-MB and LDH at T3-T5 in the control group increased,and the levels of perfusion fluid cTnT,CK,CK-MB and LDH at T4-T5 in Xuebijing group increased in comparison with those at T1 time point in the same group(all P<0.05).The levels of serum IL-1β,IL-6,TNF-αand cTnT in the Xuebijing group at T3-T5 were lower than those in the control group,and the levels of CK,CK-MB and LDH in the perfusion fluid at T2-T5 were lower than those in the control group(all P<0.05).Ultrastructure of myocardial tissue:In the control group,the ventricular muscle tissue showed moder⁃ate to severe degeneration and steatosis,the intracellular matrix was dissolved in a large area,the muscle fiber structure was disordered and broken in a large amount,the mitochondria were mostly swollen and uneven in size,and the cell mem⁃brane was dissolved and damaged.Compared with the control group,the cell structure damage in the Xuebijing group was relatively mild,the myocardial cells were slightly hypertrophic,the mitochondria were slightly swollen and uniform in size,and the muscle fibers were uniform in thickness and neatly arranged.Conclusions The addition of Xuebijing injec⁃tion to the donor heart preservation perfusate has the myocardial protective effect on the isolated beating pig heart preserved by ECMO,and its mechanism may be related to the inhibition of inflammatory response and the reduction of mitochondrial damage in cardiomyocytes.
作者
岳晓
王春华
武伟
覃冠斌
罗兰
黄强信
银世杰
YUE Xiao;WANG Chunhua;WU Wei;QIN Guanbin;LUO Lan;HUANG Qiangxin;YIN Shijie(The First Clinical Medical College,Guangxi University of Chinese Medicine,Nanning 530000,China;不详)
出处
《山东医药》
CAS
2024年第23期26-30,共5页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81960799)
广西中医药大学B类“高层次人才培育创新团队”项目(2022B007)。
关键词
血必净注射液
心脏移植
巴马小型猪
体外膜肺氧合
离体心脏
心肌保护
Xuebijing injection
heart transplantation
bama miniature pig
extracorporeal membrane oxygen⁃ation
isolated heart
myocardial protection