隔药饼灸对功能性胃肠病(肝郁脾虚证)大鼠海马和杏仁核AMPA受体亚基表达的影响

Influence of cake-separated moxibustion on expression of AMPA receptor subunits in the amygdala and hippocampus in a rat model of functional gastrointestinal disorder (liver-stagnation and spleendef ciency syndrome)

目的:探讨隔药饼灸治疗功能性胃肠病(functional gastrointestinal disorders,FGIDs)(肝郁脾虚证)中枢-氨基羟甲基恶唑丙酸(amino-3-hydroxy-5-methyl-4-isoxazole propionic acid,AMPA)受体基因的调节机制.方法:72只SD大鼠随机等分为6组,即空白组、模型组、隔药饼灸组、6-氰基-7-硝基喹喔啉-2,3-二酮(6-cyano-7-nitroquinoxaline-2,3-dione,CNQX)组、隔药饼灸+CNQX组、假手术组.除空白组外,其余5组均采用慢性束缚应激+过度疲劳+饮食失节方法造模,隔药饼灸组与隔药饼灸+CNQX组在造模前30min隔药饼灸5壮.造模结束后,运用脑立体定位仪于杏仁核(双)上微量注射-氨基羟甲基恶唑丙酸(amino-3-hydroxy-5-methyl-4-isoxazole propionic acid,AMPA)受体拮抗剂造CNQX组,加造假手术组为了评估手术创伤对模型的影响程度.采用Western blot方法检测海马CA1区和杏仁核AMPA受体亚型GluR1、GluR2的表达变化,比较各组在各区指标变化.结果:在海马CA1区和杏仁核BLA区,与模型组相比,隔药饼灸组、隔药饼灸+CNQX组和CNQX组GluR1、GluR2表达差异均有统计学意义(GluR1表达分别为1.05±0.13vs0.59±0.14,1.05±0.13vs0.33±0.08,1.05±0.13vs0.49±0.14,0.95±0.22vs0.46±0.09,0.95±0.22vs0.31±0.18,0.95±0.22vs0.47±0.13,均P<0.05;GluR2表达分别为0.33±0.08vs0.76±0.13,0.33±0.08vs1.13±0.15,0.33±0.08vs0.60±0.08,0.29±0.04vs0.46±0.08,0.29±0.04vs0.85±0.13,0.29±0.04vs0.48±0.09,均P<0.05);隔药饼灸+CNQX组GluR1表达最低,但GluR2表达最高,与CNQX组相比,隔药饼灸+CNQX组GluR2表达在海马CA1区和杏仁核区差异均有统计学意义(0.60±0.08vs1.13±0.15,0.48±0.09vs0.85±0.13,均P<0.05).结论:隔药饼灸可能通过调节FGIDs(肝郁脾虚证)模型大鼠杏仁核和海马AMPA受体,使杏仁核和海马的"兴奋-抑制"失衡得到调整,从而达到治疗肝郁脾虚证FGIDs的目的.

AIM： To investigate the influence of cake-separated moxibustion on central α-amino-hydroxymethyl oxazole propionic acid （AMPA） receptor expression in rats with functional gastrointestinal disorder （FGID） （liver-depression and spleen-deficiency syndrome）. METHODS： Seventy-two SD rats were randomly divided into six groups： a blank group, a model group, a cake-separated moxibustion group, a 6-cyano-7-nitroquinoxaline-2,3-dione （CNQX） group, a cake-separated moxibustion plus CNQX group, and a sham-operation group. Except for the blank group, the other five groups underwent chronic restraint stress ± over-fatigue ± diet disloyal modeling. Thirty minutes before modeling, cake-separated moxibustion for 5 Zhuang was applied for the cake-separated moxibustion group and cake-separated moxibustion ± CNQX group. After modeling, microinjection of AMPA receptor antagonist in the amygdala （double） was performed in the CNQX group and sham-operation group using the stereotactic positioning system to assess the effect of surgical trauma on model rats. Western blot was used to detect the expression of AMPA receptor subunits GluR1 and GluR2 in the hippocampal CA1 region and amygdala. RESULTS： Compared to the model group, GluR1 and GluR2 expression in the hippocam- pal CA1 region and amygdala BLA region wassignificantly lower in the cake-separated moxi- bustion group, cake-separated moxibustion ± CNQX group and CNQX group （GluR1 in the CA1 region： 1.05 ± 0.13 vs 0.59 ±_ 0.14, 0.33 ± 0.08, 0.49 ± 0.14, all P 〈 0.05; GluR1 in the BLA region： 0.95 ± 0.22 vs 0.46 ± 0.09, 0.31 ± 0.18, 0.47 ± 0.13, all P 〈 0.05; GluR2 in the CA1 region： 0.33 ± 0.08 vs 0.76 ± 0.13, 1.13 ± 0.15, 0.60 ± 0.08, all P 〈 0.05; G1UR2 in the BLA region： 0.29 ± 0.04 vs 0.46 ± 0.08, 0.85 ± 0.13, 0.48 ± 0.09, all P 〈 0.05）. The Cake-separated moxibustion ＋ CNQX group had the lowest expression of GluR1 and highest expression of GluR2. GluR2 expression in the hippocampal CA1 region and amygdala BLA region differed significantly between the CNQX group and cake-separated moxibustion group （0.60 ± 0.08 vs 1.13 ± 0.15, 0.48 ± 0.09 vs 0.85 ± 0.13, both P 〈 0.05）. CONCLUSION Cake-separated moxibustion can regulate amygdala and hippocampal AMPA receptor subunit gene expression in rats with FGID.