依达拉奉对急性阿维菌素中毒大鼠脑损害的保护作用及机制研究

Research about protective effects and mechanism of edaravone on brain injury in rats with acute avermectin poisoning

目的研究依达拉奉对急性阿维菌素中毒大鼠脑损害的保护作用及机制研究。方法将36只雄性SD大鼠随机分为依达拉奉治疗组(治疗组)、阿维菌素中毒组(中毒组)和对照组。采用经口灌胃方式染毒制备急性阿维菌素中毒模型,染毒后给予0.9%氯化钠注射液或0.9%氯化钠注射液+依达拉奉的不同治疗,观察三组大鼠生存状况。在染毒后72 h,以0.4%戊巴比妥钠深度麻醉后,以4%多聚甲醛灌注固定后取脑,行脑组织苏木精-伊红(HE)染色及免疫组化检测三组大鼠脑组织病理变化,观察脑组织凋亡调节蛋白Bcl-2相关X蛋白(Bax)、细胞淋巴瘤/白血病-2蛋白(Bcl-2)的表达及脑神经细胞凋亡率的变化。结果中毒组、治疗组大鼠均有病理损害;治疗组主要表现为脑神经细胞肿胀,间质水肿,血管充血,纤维紊乱,炎症细胞浸润;中毒组除上述表现外,并见多发性,局灶性神经组织破坏断裂、溶解坏死灶;对照组未见脑神经细胞形态学改变。与对照组大鼠比较,中毒组和治疗组的大鼠脑组织Bax蛋白表达增加,Bcl-2蛋白表达减少,Bax/Bcl-2比值升高,脑神经细胞凋亡率(AI)升高,差异均有统计学意义(t分别=4.15、3.34、5.07、2.06、2.96、3.61、2.54、1.95,P均<0.05);与中毒组大鼠比较,治疗组大鼠脑组织Bax蛋白表达减少,脑组织Bcl-2蛋白表达增多,Bcl-2蛋白表达减少,Bax/Bcl-2比值降低,脑神经细胞AI降低,差异均有统计学意义(t分别=3.29、4.17、2.85、2.34,P均<0.05)。结论依达拉奉可减轻急性阿维菌素中毒大鼠中毒症状,明显地减轻脑组织病理损害,通过抑制Bax蛋白表达,促进Bcl-2蛋白表达增多,明显地抑制脑神经细胞凋亡,进而对急性阿维菌素中毒大鼠的脑损害起保护作用。

Objective To study the protective effects and mechanism of edaravone on the brain injury in rats with acute avermectin（AVM） poisoning. Methods Thirty-six male sprague-dawley（SD）rats were randomly divided into the edaravone treatment group（treatment group）,the avermectin poisoning group（poisoning group） and the control group. Acute abamectin poisoning models were infected and producted by filling the stomach through the mouth. After exposure,the rats treated with saline or saline ＋ edaravone respectively. The survivals of the rats in each group for 72 hours were observed. At the 72 hours after exposure,pathological changes of brain tissue of rats were detected by HE staining and immunohisto-chemical detection after deep narcotizing with 0.4% pentobarbital sodium and perfuseing with 4% paraformaldehyde. The changes of brain nerve cell apoptosis,apoptosis regulating protein Bcl-2 and Bax expressions were analyzed. Results There were pathological damage in the poisoning group and the treatment group rats. The treatment group mainly character-ized by swelling in the brain nerve cells,interstitial edema,vascular engorgement,fiber disorder and inflammatory cells infiltration. The poisoning group showed multiple,focal neural tissue damage fracture and dissolve focal necrosis besides the above performances.The control group did not find cranial nerve cell morphological changes. Compared with the control group,the Bax proteins in brain tissue of poisoning group and treatment group were significantly increased,Bcl-2 protein expressions were significantly decreased,Bax/Bcl-2 ratio were significantly increased,the brain cells apoptosis rate（AI） were significantly increased（t =4.15,3.34,5.07,2.06,2.96,3.61,2.54,1.95,P〈0.05）. But compared with the rats in the poisoning group,the expression of Bax protein of treatment group was significantly decreased in brain tissue,the expression of Bcl-2 protein was significantly increased,Bax/Bcl-2 ratio was significantly decreased,the nerve cells AI was significantly decreased（t =3.29,4.17,2.85,2.34,P〈0.05）. Conclusion Edaravone could reduce the poisoning symptoms of the rats with AVM poisoning and significantly reduce pathological damage by inhibiting the expression of Bax protein. It promotes the expression of Bcl-2 protein through inhibiting Bax protein,significantly inhibit the apoptosis of nerve cells and protects brain that damage of abamectin acute poisoning.