大建中汤对脾阳虚疼痛大鼠COX-2mRNA及蛋白和CaMKⅡmRNA的影响

Effect of Da Jianzhong Decoction on Cyclooxygenase-2 m RNA and Protein and Calmodulin-dependent Protein Kinase Ⅱ m RNA in Spleen-yang Deficiency Pain Rats

【目的】观察大建中汤对脾阳虚疼痛大鼠脑组织环氧合酶-2(COX-2)m RNA及蛋白和钙调蛋白依赖的蛋白激酶Ⅱ(Ca MKⅡ)m RNA的影响,探讨大建中汤对脾阳虚疼痛的作用机理。【方法】将SD大鼠40只随机分为正常组,模型组,大建中汤高、低剂量组(剂量分别为3、0.75 g·kg^(-1)·d^(-1))各10只,采用综合造模法复制脾虚动物模型,根据实验分组以灌胃形式连续给药15 d后,断颈处死大鼠并立即取脑组织。采用Real-time PCR法检测脑组织COX-2 m RNA、Ca MKⅡm RNA表达的变化;免疫组织化学法检测COX-2蛋白表达变化。【结果】与正常组比较,脾阳虚模型组大鼠大脑COX-2 m RNA及蛋白表达显著增加(P<0.01),Ca MKⅡm RNA表达显著降低(P<0.01);大建中汤高、低剂量组COX-2 m RNA与蛋白表达较模型组显著下降(P<0.01),Ca MKⅡm RNA表达较模型组显著升高(P<0.01)。【结论】大建中汤可通过抑制脾阳虚疼痛大鼠脑组织中COX-2 m RNA及蛋白表达,提高Ca MKⅡm RNA表达来达到治疗疼痛的目的。

Objective To observe the effect of Da Jianzhong Decoction on cerebral cyclooxygenase-2（COX-2）m RNA and protein and calmodulin-dependent protein kinase II（Ca MKⅡ） m RNA in spleen-yang deficiency pain rats,so as to investigate the therapeutic mechanism of Da Jianzhong Decoction for spleen-yang deficiency pain. Methods Forty SD rats were randomly divided into normal group,model group,and high- and low- dose Da Jianzhong Decoction groups（3,0.75 g·kg-1·d-1）,10 in each group. Spleen deficiency rat model was established by comprehensive modeling methods. After the rats in various groups were given gastric administration of Chinese medicine for 15 continuous days, rat cervical dislocation was performed for the sampling of the brain tissue. The expression levels of COX-2 m RNA and Ca MK Ⅱ m RNA in brain tissue were detected by real time polymerase chain reaction（RT-PCR）. The COX-2 protein expression was determined by immunohistochemistry.Results Compared with the normal group,m RNA and protein expression levels of COX-2 in rat brain tissue of spleen-yang deficiency model group were increased（P﹤0.01）,and Ca MKⅡ m RNA expression was decreased（P〈0.01）. Compared with the model group,COX-2 m RNA and protein expression levels were decreased（P〈0.01）and Ca MKⅡ expression was increased（P〈0.01）in high- and low-dose Da Jianzhong Decoction groups（P〈0.01）.Conclusion Da Jianzhong Decoction can suppress cerebral COX-2 m RNA and protein expression, and up-regulate Ca MKⅡm RNA expression,which probably contribute to its therapeutic mechanism for relieving the pain.