黄连解毒汤合定痫丸治疗阳痫大鼠的实验研究

Experimental study of Huanglian Jiedu decoction combined with Dingxian Pill on Yangxian Epileptic Rats

目的观察黄连解毒汤合定痫丸(汤剂)对阳痫大鼠的抗痫作用。方法以高脂饲料和制附子煎液预饲养并采用匹罗卡品注射法制成阳痫大鼠模型,以黄连解毒汤合定痫丸(汤剂)灌胃治疗(阳痫中药组),同时设正常组、阳痫组和阳痫中药反治组进行对照,每组各9只观测癫痫发作潜伏期、到达癫痫持续状态(SE)时间等症状学指标,高效液相色谱法检测脑内谷氨酸(Glu)、天门冬氨酸(Asp)、γ-氨基丁酸(GABA)、甘氨酸(Gly)、丙氨酸(Ala)等5种氨基酸类神经递质的含量,伊红-苏木素(HE)切片观察肺、肾、肝、海马、脑皮质的病理变化,免疫组化法测定脑内caspase-3表达。结果阳痫组、阳痫中药反治组、阳痫中药组大鼠均出现4级以上癫痫发作。癫痫发作潜伏期以阳痫组最长,阳痫中药组次之,阳痫中药反治组最短;到达SE时间以阳痫中药组最长,阳痫中药反治组次之,阳痫组最短。各致痫组大鼠脑内5种氨基酸含量较正常组均降低。阳痫组Glu、Asp与正常组比较差异有统计学意义(P<0.05,P<0.01),GABA、Gly、Ala与正常组比较差异无统计学意义(P>0.05);阳痫中药反治组中各种氨基酸与正常组比较差异有统计学意义(P<0.05,P<0.01);阳痫中药组Glu、Asp、GABA、Gly与正常组比较差异有统计学意义(P<0.05,P<0.01),Ala与正常组比较差异无统计学意义(P>0.05)。HE切片显示,正常组大鼠各组织基本正常,其余组大鼠肺、肾、肝、海马、脑皮质等组织均出现明显病理损害。阳痫组和阳痫中药反治组的脑内caspase-3水平高于正常组,其中,阳痫中药反治组升高较明显,而阳痫中药组则略低于正常组,但比较差异均无统计学意义(P>0.05)。结论黄连解毒汤合定痫丸可能能推迟阳痫大鼠SE的到达时间,改变部分脑内氨基酸神经递质水平,并减少神经元凋亡,但作用程度较轻。

Objective To explore anti - epileptic effects of Huanglian Jiedn decoction combined with Dingxian Pill on pilocarpine -yangxian epileptic rats. Methods The yangxian epileptic rat models were established by intraperitoneal injection of pilocarpine after being pretreated with high fat diet and Radix Aconiti Lateralis Preparata decoction. Then, they were treated with Huanglian Jiedu decoction and Dingxian Pill, which were referred as the yangxian Chinese medicine （CM） group. At the same time, the normal group, the yangxian group, and the yangxian CM converse therapy group were established to serve as controls. Symptomatological indexes like time spots of seizure latency period and status epilepticus （SE） were recorded; concentrations of five amino acid neurotransmitters in the brains including glutamic acid （Glu）, aspartie acid （Asp）, gamma- aminobutyric acid（GABA） , glycine （Gly）, and alanine （Ala） were detected by high- performance liquid chromatography; pathological changes including the lung, the kidney, the liver, the hippocampus and the cortex were observed by HE staining ; the expression of caspase- 3 in brains were examined by immunohistochemisty. Results Class 4 - 5 seizures were observed in all the rats in the yangxian group, the yangxian CM converse therapy group, and the yangxian CM group. The seizure latency period of the yangxian group was the longest, followed by the yangxian CM group, and the yangxian CM converse therapy group was the shortest. The latency time of SE of the yangxian CM group was the longest, and the yangxian group wasthe shortest, whereas the yangxian CM converse therapy group was in the midst. All the concentrations of the five amino acids in brains of epileptic rats decreased when compared with the normal group. Compared with the nor- mal group, there were significant decrease in the concen-trations of Glu and Asp of the yangxian group, all the amino acids of the yangxian CM converse therapy group, and Glu, Asp, GABA and Gly of the yangxian CM group （ P 〈0.05, P 〈0.01）. However, GABA, Gly and Ala of the yangxian group, and Ala of the yangxian CM group were no statistically significant decline （ P 〉 0.05）. HE stain in- dicated severe damage in the lung, the kidney, the liver, the hippocampus and the cortex of all the epileptic rats, while all the tissues mentioned above of the normal group were normal. The expression of caspase - 3 of the yangxian group and the yangxian CM converse therapy group, which had more obvious elevation, ascended, while the yangxian CM group mildly descended as compared with the normal group, but there was no statistical difference among all the groups. Conclusiou Huanglian Jiedu decoction combined with Dingxian Pill would delay the occurrence of SE of the yangxian epileptic rats, regulate part of the concentrations of the amino acid neurotransmitters in brains, and prohibit apoptosis of neurons.