蜜蜂蜂毒涂膜剂经皮给药对实验性脑血栓大鼠的改善作用

Improvement Effects of Bee Venom Plastics on Experimental Cerebral Thrombosis in Rats

目的:研究蜜蜂蜂毒(BV)涂膜剂经皮给药对实验性脑血栓大鼠的改善作用。方法:取SD大鼠96只,随机分为假手术组(生理盐水)、模型组(涂膜剂空白基质)、尼莫地平组(阳性药物,4.00 mg/kg)和BV涂膜剂低、中、高剂量组(1.67、3.33、6.67 mg/kg),每组16只,假手术组和尼莫地平组大鼠灌胃给药,模型组和BV涂膜剂各剂量组大鼠经皮涂抹给药。连续给药5 d后,采用结扎右颈外动脉和翼颚动脉,同时颈内动脉注射复合血栓诱导剂的方法建立实验性脑血栓模型。分别测定大鼠右脑/左脑的湿质量比以考察梗死侧脑组织水肿程度;采用紫外分光光度法测定大鼠左右半脑中伊文思蓝(EB)含量以考察脑血管通透性;检测大鼠血液流变学和凝血功能指标;采用苏木精-伊红染色法观察大鼠脑组织病理学变化,并计数存活神经细胞数。结果:经与假手术组各指标比较显示,大鼠脑血栓模型成功建立。与模型组比较,BV涂膜剂各剂量组大鼠右侧(梗死侧)脑组织蓝染面积明显缩小,右脑/左脑湿质量比和右脑组织中EB含量均显著降低(P<0.05或P<0.01);BV涂膜剂各剂量组大鼠全血黏度和卡松黏度,以及BV涂膜剂中、高剂量组大鼠血浆黏度均显著降低(P<0.01);BV涂膜剂中剂量组大鼠凝血酶原时间和活化部分凝血活酶时间显著延长(P<0.01);BV涂膜剂各剂量组大鼠脑组织病理学变化明显减轻,神经细胞排列较为整齐,结构清晰,核仁清晰,胞膜完整,存活神经细胞数显著增加(P<0.01)。结论:BV涂膜剂能减轻脑血栓模型大鼠的脑水肿程度,抑制其脑血管通透性,改善脑血栓形成后大鼠血液流变学和凝血功能指标,减轻其缺血后神经细胞损伤。

OBJECTIVE:To study the improvement effects of Bee venom(BV)plastics on experimental cerebral thrombosis in rats.METHODS:Totally 96 SD rats were randomly divided into sham operation group(normal saline),model group(plastics blank matrix),Nimodipine group(positive drug,4.00 mg/kg)and BV plastics low-dose,medium-dose,high-dose groups(1.67,3.33,6.67 mg/kg),with 16 rats in each group.Rats in sham operation group and Nimodipine group were given medicine intragastrically,while rats in model group and BV plastics groups were given medicine by transdermal smearing.After 5 days of continuous administration,the experimental cerebral thrombosis model was established by ligating the right external carotid artery and pterygomandibular artery,and injecting compound thrombus inducer into the internal carotid artery.The wet mass ratio of right brain to left brain was measured to investigate the degree of brain edema on the infarcted side.The content of Evans blue(EB)in the left and right hemispheres of rats was determined by ultraviolet spectrophotometry to investigate the cerebral vascular permeability.Blood rheology and coagulation function indicators of rats were measured.The pathological changes of brain tissue in rats were observed by HE staining,and the number of survival neuron cells was counted.RESULTS:Compared with the indexes of sham operation group,the cerebral thrombosis model was established successfully.Compared with model group,the area of blue staining in the right brain(infarcted side)of rats in BV plastics groups was significantly reduced,and the right brain/left brain wet mass ratio and the content of EB in the right brain tissue were significantly reduced(P<0.05 or P<0.01).The whole blood viscosity and Casson viscosity of rats in BV plastics groups,and the plasma viscosity of rats in BV plastics medium-dose and high-dose groups decreased significantly(P<0.01).PT and APTT of rats were prolonged significantly in BV plastics medium-dose group(P<0.01).The pathological changes of brain tissue in rats in BV plastics groups were significantly alleviated.The arrangement of neuron cells was more orderly,the shape and structure of cells were clear,the nucleolus was clear,the membrane was intact,and the number of survival neuron cells was significantly increased(P<0.01).CONCLUSIONS:BV plastics can alleviate brain edema,inhibit cerebral vascular permeability,improve hemorheology and coagulation function indicators of rats after the formation of cerebral thrombosis,and alleviate nerve cell injury after ischemia.