蒙药朝伦雄胡-5对癫痫持续状态大鼠模型中VEGF表达影响的研究

Study on the Effect of Mongolian Medicine Zhaolun Xionghu-5 on VEGF Expression in Rat Model with Status Epilepticus

摘要: 目的：通过氯化锂–毛果芸香碱建立大鼠癫痫持续状态模型，探究癫痫持续状态大鼠模型中不同时间点VEGF的表达变化，并探讨蒙药朝伦雄胡-5对VEGF表达的影响。方法：第一部分：选取90只6~8周龄健康雄性SD大鼠，随机分为造模成功组、给药对照组和生理盐水对照组，每组30只。造模成功组与给药对照组通过腹腔注射氯化锂–毛果芸香碱制作大鼠癫痫持续状态模型，生理盐水对照组给予同等剂量的生理盐水处理，造模完成后应用qRT-PCR和Western blot方法检测VEGF转录水平及蛋白水平的表达情况。第二部分：选取40只6~8周龄健康雄性SD大鼠，随机分为蒙药组和对照组。制作癫痫大鼠模型前3 d，蒙药组给予蒙药朝伦雄胡-5灌胃处理，对照组给予同等剂量的生理盐水灌胃，3 d后对蒙药组和对照组通过腹腔注射氯化锂–毛果芸香碱制作大鼠癫痫持续状态模型。造模完成后应用qRT-PCR和Western blot方法检测VEGF转录水平及蛋白水平的表达情况。结果：第一部分：造模成功组VEGF mRNA表达较给药对照组和生理盐水对照组相比显著升高(P < 0.05)；给药对照组与生理盐水对照组VEGF mRNA表达水平无明显差异。且造模成功组VEGF mRNA随着时间的增长而逐渐升高，给药对照组、生理盐水对照组VEGF mRNA不随时间而变化。造模成功组较另外2组相比，VEGF蛋白表达水平显著增高(P < 0.05)；给药对照组与生理盐水对照组VEGF蛋白表达水平无明显差异。而且3组的VEGF蛋白变化均无明显的时间依赖性。第二部分：蒙药组VEGF mRNA与对照组相比，表达显著降低(P < 0.05)，并且随着时间的增长，VEGF不断降低；蒙药组较造模成功组VEGF蛋白表达显著降低(P < 0.05)，但并不存在时间依赖性。结论：1) 在氯化锂–毛果芸香碱诱导的癫痫持续状态大鼠海马中的VEGF mRNA的表达是升高的。2) 在氯化锂–毛果芸香碱诱导的癫痫持续状态大鼠海马中的VEGF蛋白的表达是升高的。3) 蒙药朝伦雄胡-5可诱导癫痫持续状态大鼠海马中的VEGF mRNA表达下降，并且随着时间的增长而不断降低。4) 蒙药朝伦雄胡-5在癫痫持续状态模型中从蛋白水平上可部分抑制VEGF表达升高。

Abstract: Objective: To explore the changes in VEGF expression at different time points in the rat model of status epilepticus by lithium chloride-rutin and explore the effect of Mongolian medicine Zhaolun Xionghu-5 on VEGF expression. Methods: Part I: 90 healthy male rats aged 6~8 weeks were randomly divided into successful modeling group, drug administration control group and normal saline control group, with 30 rats in each group. The rats in the successful modeling group and the control group were intraperitoneally injected with lithium chloride-rutin to establish the rat model of status epilepticus. The rats in the normal saline control group were treated with the same dose of normal saline. After the modeling, qRT-PCR and Western blot were used to detect the expression of VEGF transcription and protein levels. Part II: 40 healthy male rats aged 6~8 weeks were randomly divided into Mongolian medicine group and control group. Three days before the establishment of the rat model of status epilepticus, the Mongolian medicine group was given Mongolian medicine Zholun Xionghu-5 by gavage, and the control group was given the same dose of normal saline by gavage. Three days later, the Mongolian medicine group and the control group were intraperitoneally injected with lithium chloride-rutin to establish the rat model of status epilepticus. After the modeling, qRT-PCR and Western blot were used to detect the expression of VEGF transcription and protein levels. Results: Part I: The mRNA expression of VEGF in the successful modeling group was significantly higher than that in the administration control group and the normal saline control group (P < 0.05). There was no significant difference in VEGF mRNA expression between the administration control group and the normal saline control group. Moreover, VEGF mRNA in the successful modeling group increased gradually with the increase of time, while VEGF mRNA in the administration control group and normal saline control group did not change with time. Compared with the other two groups, the protein expression of VEGF in the successful modeling group was significantly increased (P < 0.05). There was no significant difference in VEGF protein expression between the administration control group and the normal saline control group. Moreover, the changes in VEGF protein in the three groups were no time dependence. Part II: Compared with the control group, the expression of VEGF mRNA in the Mongolian medicine group was significantly decreased (P < 0.05), and with the growth of time, VEGF continued to decrease; the expression of VEGF protein in Mongolian medicine group was significantly lower than that in successful modeling group (P < 0.05), but there was no time dependence. Conclusion: 1) VEGF mRNA expression was increased in hippocampus of rats with status epilepticus induced by lithium chloride-rutin. 2) The expression of VEGF protein in hippocampus of rats with status epilepticus induced by lithium chlorine-rutin was increased. 3) Mongolian medicine Zhaolun Xionghu-5 can induce the decrease of VEGF mRNA expression in hippocampus of rats with status epilepticus, and it decreases with time. 4) Mongolian medicine Zhaolun Xionghu-5 can partially inhibit elevated VEGF expression at the protein level in the model of status epilepticus.