七氟醚调控CNCA/Ras/ERK信号通路减轻脑出血大鼠神经损害的机制研究

EFFECTS OF SEVOFLURANE POST-TREATMENT ON NERVER DAMAGE IN RATS WITH INTRACEREBRAL HEMORRHAGE

目的 研究七氟醚(sevoflurane,Sev)后处理对脑出血(intracerebral hemorrhage,ICH)大鼠CNCA/Ras/ERK信号通路的影响,探究其减轻脑出血后神经损害的可能作用机制。方法 选取健康雄性SD大鼠36只,应用胶原酶法制作大鼠脑出血模型,采用随机数字表法分为三组:假手术组(sham组)、脑出血组(ICH组)、脑出血七氟醚后处理组(ICH+Sev组),每组12只。ICH+Sev组大鼠于脑出血后立即给予体积分数2.4%七氟醚和氧气持续吸入30 min,氧流量1L/min,sham组和ICH组大鼠持续吸入同样时长及同样流量的氧气。于术后24 h应用改良神经功能评分mNSS评估大鼠神经功能,采用TUNEL法检测脑组织神经元凋亡情况、通过蛋白印迹分析测定脑组织CACNα1d、RasGRP、ERK等蛋白表达,采用ELISA法测定血清中炎症因子IL-6及TNF-α的表达情况。结果与sham组相比,ICH组mNSS评分、细胞凋亡数、IL-6和TNF-α表达以及CACNα1d和ERK的蛋白表达量均上升,而RasGRP的蛋白表达量下降(均P<0.05);与ICH组相比,ICH+Sev组mNSS评分、细胞凋亡数、IL-6和TNF-α的表达以及CACNα1d和ERK的蛋白表达量均下降,而RasGRP的蛋白表达量上升(均P<0.05)。结论 七氟醚后处理对脑出血大鼠具有脑保护作用,该作用可能通过调控CNCA/Ras/ERK信号通路起到抑制神经元凋亡、减轻炎症,从而达到改善神经功能障碍的作用。

Objective To investigate the effect of sevoflurane(Sev) post-treatment on CNCA/Ras/ERK signaling pathway in rats with intracerebral hemorrhage(ICH),and to explore its possible mechanism of reducing nerve damage after intracerebral hemorrhage.Methods Thirty-six healthy male SD rats were selected,and the rat model of intracerebral hemorrhage was made by the collagenase method.They were divided into three groups by random number table method:sham operation group(sham group),intracerebral hemorrhage group(ICH group),sevoflurane post-treatment group for intracerebral hemorrhage(ICH+Sev group),12 animals in each group.Rats in ICH+Sev group were given volume fraction of 2.4% sevoflurane and oxygen for 30 min immediately after intracerebral hemorrhage,and the oxygen flow rate was 1 L/min.Rats in sham group and ICH group continued to inhale oxygen for the same duration and flow.At 24 hours after operation,the modified neurological function score mNSS was used to evaluate the neurological function of the rats,the TUNEL method was used to detect the apoptosis of neurons in the brain tissue,the protein expressions of CACNα1 d,RasGRP,ERK in the brain tissue were determined by Western blot analysis,and the expression ofinflammatory factors IL-6 and TNF-α in serum were determined by ELISA.Results Compared with the sham group,the mNSS score,the number of apoptosis,the expressions of IL-6 and TNF-α,and the protein expressions of CACNα1 d and ERK in the ICH group all increased,while the protein expression of RasGRP decreased(P<0.05);Compared with the ICH group,the mNSS score,the number of apoptotic cells,the expressions of IL-6 and TNF-α,and the protein expressions of CACNα1 d and ERK in the ICH+Sev group all decreased,while the protein expression of RasGRP increased(P<0.05).Conclusion Post-treatment with sevoflurane has a brain protective effect on rats with intracerebral hemorrhage,which may inhibit neuronal apoptosis and reduce inflammation by regulating the CNCA/Ras/ERK signaling pathway,thereby improving neurological dysfunction.