盐酸戊乙奎醚通过NF-κB信号通路对胎鼠脑缺血-再灌注损伤的影响

Penehyclidine hydrochloride induces cerebral ischemia-reperfusion injury of fetal rats through inhibition of NF-κB signaling pathway

目的研究盐酸戊乙奎醚对宫内窘迫致胎鼠脑缺血-再灌注（IR）损伤的保护作用。方法足月胎鼠80只,体重4.52~4.81 g,采用随机数字表法分为四组：假手术组（S组）、盐酸戊乙奎醚对照组（S＋P组）、IR组和盐酸戊乙奎醚治疗组（IR＋P组）,每组20只。采用钳夹孕鼠两侧子宫角血管的方法建立宫内窘迫模型,IR＋P组孕鼠建模前30 min肌注盐酸戊乙奎醚2 mg/kg,S组在假手术前予孕鼠肌注等量生理盐水,S＋P组在假手术前予孕鼠肌注等量盐酸戊乙奎醚。再灌注12 h后,剖宫取出胎鼠并断头处死,行外周血血气分析;采用TTC染色法观察胎鼠脑梗死体积并计算脑梗死体积百分比;采用HE染色观察胎鼠脑组织病理改变;采用ELISA法检测脑组织中TNF-α及IL-6的浓度;采用RT-PCR法检测脑组织中NF-κB mRNA表达;采用Western blot法检测胎鼠脑组织中NF-κB p65蛋白表达。结果 IR组和IR＋P组p H和Pa O2明显低于S组和S＋P组,IR＋P组p H和Pa O2明显高于IR组（P〈0.05）。IR组和IR＋P组Pa CO2、Lac、脑梗死体积及梗死体积百分比、TNF-α及IL-6浓度、NF-κB mRNA表达水平、NF-κB p65蛋白水平均明显高于S组和S＋P组（P〈0.05）,IR＋P组上述各指标均明显低于IR组（P〈0.05）。IR＋P组脑组织病理学损伤明显轻于IR组。结论盐酸戊乙奎醚预处理可减轻宫内窘迫致胎鼠脑缺血-再灌注损伤,其作用机制可能与抑制脑组织NF-κB信号通路活性有关。

Objective To investigate the effects of penehyclidine hydrochloride（ PHCD） on cerebral ischemia-reperfusion injury induced by intrauterine distress in fetal rats. Methods Eighty mature fetal rats weighing 4. 52-4. 81 g were randomly divided into four groups（ n = 20） ： sham operation group（ group S）,PHCD control group（ group S ＋ P）,cerebral IR group（ group IR）,PHCD treatment group（ group IR ＋P）. Fetal rat intrauterine distress model was set up by clamping bilateral uterine horn vessels of pregnant rats. PHCD 2 mg / kg was injected in pregnant rat＇s gluteus at 30 min before intrauterine distress model was set up in group IR ＋ P,the same volume saline was injected in pregnant rat＇s gluteus before shame operation in group S,the same volume PHCD was injected in pregnant rat＇s gluteus before shame operation in group S ＋ P. Fetal rats were decapitated at 12 h after the reperfusion,the peripheral blood of fetal rats was detected by blood gas analysis（ including PH,Pa O2,Pa CO2,Lac）; the infarct volume and the infarct volume fraction were detected by TTC staining; pathological changes in lung tissue were observed by HE staining; the TNF-α,IL-6 content in the brain were detected by ELISA; the expression of NF-κB mRNA was detected by quantitative Real-time PCR,the expression of NF-κB p65 protein was detected by Western-blotting. Results The blood PH,Pa O2 in group IR and IR ＋ P were lower than group S and S ＋ P,the blood PH,Pa O2 in group IR ＋ P was higher than group IR. Compared with group S and group S ＋ P,the blood Pa CO2,Lac,the infarct volume and the infarct volume fraction,the concentration of TNF-α and IL-6,the expression of NF-κB mRNA and protein were significantly increased in group IR and IR ＋ P（ P〈0. 05）,and those in group IR ＋ P were lower than group IR（ P〈0. 05）. The pathological changes in brain tissue were significantly attenuated in group IR ＋ P（ P〈0. 05）. Conclusion Pretreatment with PHCD could attenuate cerebral ischemia-reperfusion injury of fetal rats induced by intrauterine distress. The mechanisms could relate to the inhibition of NF-κB signaling pathway in brain tissues.