同病异治对戊四氮点燃大鼠海马区谷氨酸代谢通路的影响

Effects of Heterotherapy for Homopathy on the Metabolism Path of Glutamate in the Pentylenetetrazol-kindled Seizure Rats′ Hippocampus

目的观察并比较癫痫"从肝论治"复方柴胡疏肝汤和"从痰论治"定痫丸对戊四氮(pentylenetetrazole,PTZ)点燃大鼠海马区谷氨酸代谢通路的影响,探讨同病异治理论的分子机制。方法以亚惊厥剂量PTZ腹腔注射大鼠建立慢性点燃癫痫模型。完全点燃大鼠24只,随机分为4组,分别为模型组、丙戊酸钠组、定痫丸组和柴胡疏肝汤组,并分别给予生理盐水、丙戊酸钠、定痫丸和柴胡疏肝汤灌胃干预;对照组大鼠给予生理盐水腹腔注射和灌胃干预。连续灌胃4周后,应用HPLC荧光法检测大鼠海马谷氨酸(gluta-mate,Glu)含量;Western blot技术观察谷氨酸转运体-1(glutamate transporter-1,GLT-1)蛋白表达水平;谷氨酰胺合成酶检测试剂盒检测谷氨酰胺合成酶(glutamine synthetase,GS)活性。结果与对照组比较,模型组大鼠海马区Glu含量显著升高、GLT-1蛋白表达及GS活性显著降低(P<0.01);与模型组比较,各用药组大鼠海马区Glu含量均显著降低,GLT-1蛋白表达及GS活性均显著升高(P<0.01);两中药复方组比较,柴胡疏肝汤组大鼠海马区Glu含量降低及GS活性升高较定痫丸组更显著(P<0.01),而两组GLT-1蛋白表达,差异无统计学意义(P>0.05)。结论 "从肝论治"复方柴胡疏肝汤和"从痰论治"定痫丸均能降低PTZ点燃大鼠海马区Glu含量,提高GLT-1蛋白表达及GS活性,说明两复方均能通过调节Glu代谢通路影响脑内Glu水平;而柴胡疏肝汤降低癫痫大鼠海马区Glu含量、上调GS活性效果优于定痫丸,提示柴胡疏肝汤和定痫丸对Glu代谢通路的调节存在不同作用靶点,这可能是癫痫同病异治理论的分子机制之一。

Objective To investigate and compare the effects of Compound Chaihu Shugan Decoction （CHSGD, ＂treatment from Gan＂ ） and Dingxian Pill （ DXP, ＂treatment from the sputum＂ ） on the metabolism path of glutamate in the pentylenetetrazol-kindled seizure rats＇ hippocampus, thus exploring the molecular mechanism of ＂heterotherapy for homopathy＂. Methods A chronic kindling seizures rat model was established by intraperitoneal injecting pentylenetetrazol （PTZ）. Totally 24 fully kindled seizure rats were randomized into four groups, i.e., the model control group, the Sodium Valproate （VPA）group, the DXP group, and the CHSGD group. They were respectively treated with normal saline, VPA, CHSGD, and DXP, respectively. Rats in the control group were treated with normal saline by peritoneal injection and by gastrogavage. After intragastric administration for 4 successive weeks, the glutamate （Glu）levels in the hippocampus were detected by high performance liquid chromatography （HPLC）. The expressions of glutamate transporter-1 （GLT-1）proteins were detected by Western blot. The activity of glutamine syn- thetase （GS）was detected by using GS detection kit. Results Compared with the control group, the content of Glu in the model group significantly increased, and the expression of GLT-1 and the activity of GS significantly decreased （ P 〈0.01 ）. Compared with the model group, the content of Glu in each medication group significantly decreased, and the protein expression of GLT-1 as well as the activity of GS significantly increased （P 〈0.01 ）. But when compared between the CHSGD group and the DXP group, the content of Glu was lower and the activity of GS was higher in the CHSGD group than in the DXP group （P 〈0.01 ）, while there was no statistical difference in the expression of GLT-1 between the two groups （ P 〉0.05）. Conclusions CHSGD （＂treatment from Gan＂ ）and DXP （＂treatment from the sputum＂ ）could both decrease the level of Glu and raise the expression of GLT-1 and the activity of GS, indicating that CHSGD and DXP both could regulate the metabolism path of Glu to affect the level of the Glu in the brain. But the effects of CHSGD were superior to those of DXP in decreasing the content of Glu and up-regulating the activity of GS, suggesting that there were some different effects targets between the two compounds on the metabolism path of Glu, which may be one of possible molecular mechanisms for treating epilepsy by heterotherapy for homopathy.