摘要
目的探讨β受体阻滞剂(艾司洛尔)对脓毒症大鼠的心肌保护作用及对Toll样受体4(Toll-like receptor4,TLR4)炎症通路的影响。方法选取雄性Wistar大鼠60只,采用随机数字法随机分为假手术组(Shame组)、脓毒症组(CLP组)、艾司洛尔组(CLP+ES组)、TLR4抑制剂组(CLP+TAK-242组)每组15只。Shame组实施盲肠探查术,CLP组、CLP+ES组与CLP+TAK-242组通过盲肠结扎穿孔法(CLP法)建立脓毒症模型;CLP+ES组于CLP术后12 h给予腹腔注射艾司洛尔稀释液20 mg/kg;CLP+TAK-242组于上述相同时间点给予腹腔注射TAK-2423 mg/kg;Shame组与CLP组给予等量生理盐水。各组均于术后24 h处死大鼠,采集并处理标本。取心肌组织行苏木精-伊红染色观察大鼠心肌病理学变化;采用免疫组织化学法观察心肌组织中TLR4、髓样分化蛋白88(myeloid differentiation factor88,MyD88)及核转录因子-κB(nuclear factor-κB,NF-κB)的表达;采用马松染色(masson染色)观察心肌组织纤维化程度的表达;采用蛋白质免疫印迹试验(western blot)检测TLR4、MyD88、NF-κB以及天冬氨酸特异性半胱氨酸蛋白酶-1(aspartic acid specifi c cysteine protease 1,caspase-1)蛋白表达;取腹主动脉血采用免疫酶联免疫吸附试验(enzyme-linked immuno sorbent assay,ELISA)检测血清心肌肌钙蛋白(I cardiac troponin-I,cTn-I)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)的水平。结果与Shame组相比,CLP组心肌病理损伤、纤维化以及炎症细胞浸润明显加重,心肌损伤指标cTn-I与炎症介质TNF-α、IL-6、IL-1β水平明显升高[(8.70±0.22)vs.(4.41±0.31),(445.57±9.13)vs.(219.60±5.52),(165.55±2.18)vs.(93.47±3.37),(124.12±2.59)vs.(67.63±6.04),均P<0.05];与CLP组相比,CLP+ES组与CLP+TAK=242组心肌损伤显著减轻,炎症递质水平明显降低[(5.38±0.18)与(5.37±0.13)vs.(8.70±0.22),(322.73±7.63)与(300.58±17.47)vs.(445.57±9.13),(121.28±5.4
Objective To investigate the protective effect ofβ-blocker(esmolol)on myocardia and toll-like receptor 4(TLR4)infl ammatory pathway in septic rats.Methods Sixty male Wistar rats were randomly(random number)divided into the shame group,sepsis group(CLP group),esmolol group(CLP+ES group)and TLR4 inhibitor group(CLP+TAK-242 group)with 15 rats in each group.Cecal exploration was performed in the shame group,and cecal ligation and perforation(CLP)was performed in the CLP group,CLP+ES group and CLP+TAK-242 group.The CLP+ES group received intraperitoneal injection of esmolol diluent 20 mg/kg 12 h after CLP.The CLP+TAK-242 group was given intraperitoneal injection of TAK-2423 mg/kg at the same time point as above.The shame group and CLP group were given the same amount of normal saline.Rats in all groups were sacrificed 24 h after operation,and the samples were collected and processed.The pathological changes of myocardium were observed by hematoxylin-eosin staining.The expression of TLR4,myeloid differentiation protein 88(MyD88)and nuclear factor-κB(NF-κB)in myocardial tissue were observed by immunohistochemistry.Masson staining was used to observe the expression of fi bers and infl ammatory factors in myocardial tissue.The protein expressions of TLR4,MyD88,NF-κB and aspartic acid specifi c cysteine protease 1(caspase-1)were detected by Western blot.Serum levels of cardiac troponin I(cTn-I),tumor necrosis factorα(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β)were detected by enzyme-linked immunosorbent assay(ELISA).Results Compared with the shame group,myocardial injury,fi brosis and infl ammatory cell infiltration were significantly aggravated in the CLP group,and the levels of myocardial injury index cTn-I and inflammatory mediators TNF-α,IL-6 and IL-1βwere significantly increased[(8.70±0.22)vs.(4.41±0.31),(445.57±9.13)vs.(219.60±5.52),(165.55±2.18)vs.(93.47±3.37),(124.12±2.59)vs.(67.63±6.04),all P<0.05].Compared with the CLP group,myocardial injury was signifi cantly reduced in the CLP+ES group
作者
迪丽热巴·吐尔逊
杨春波
丁琼莉
于湘友
Dilireba·tuerxun;Yang Chunbo;Ding Qiongli;Yu Xiangyou(Department of Critical Care Medicine,The First Affiliated Hospital of Xinjiang Medical University,Xinjiang,830054,China)
出处
《中华急诊医学杂志》
CAS
CSCD
北大核心
2022年第10期1353-1360,共8页
Chinese Journal of Emergency Medicine
基金
国家自然科学基金(82160360)。
