平性药桃仁对寒热不同血瘀证大鼠cAMP-PKA信号通路的影响

Study on the Regulation Role of Semen Persicae to cAMP-PKA Signal Pathway in the Rats with Cold and Heat Blood Stasis Syndrome

目的:观察平性药桃仁对瘀热互结和寒凝血瘀两种证型大鼠cAMP-PKA信号通路的影响,探讨其双向适用的作用机理。方法:60只雄性SD大鼠随机分为正常对照组、模型组、桃仁组、丹参组、川芎组,每组12只。各中药组灌胃给予相应药液,正常对照组和模型组灌胃给水,2次/d,每次20 mL/kg,连续7 d。瘀热互结证和寒凝血瘀证分2次进行实验。瘀热互结证除正常对照组外,其余各组大鼠先经腹腔注射10%角叉菜胶5 mL/kg,1次/d,连续给药3d;末次注射角叉菜胶24 h后,再于大鼠皮下注射20%干酵母悬液10 mL/kg造成瘀热互结证大鼠模型。寒凝血瘀证除正常对照组外,其余各组大鼠置于特制鼠笼内,然后置于-(18±2)℃冰柜,2 h/次,2次/d,连续7 d,造成寒凝血瘀证大鼠模型。瘀热互结证和寒凝血瘀证分别于末次造模后6、12 h,腹主动脉取血5 mL和并取腹主动脉。采用Elisa法检测血浆中cAMP浓度采用Westernblot法检测腹主动脉PKA蛋白表达。结果:平性药桃仁在瘀热互结证中能降低瘀热互结证大鼠血浆cAMP浓度(P<0.01),抑制PKA蛋白表达(P<0.05);在寒凝血瘀证中能升高寒凝血瘀证大鼠血浆cAMP浓度(P<0.01),增强PKA蛋白表达(P<0.01),对cAMP-PKA信号通路具有双向调节的作用。结论:平性药桃仁在瘀热互结证和寒凝血瘀证两种不同的内环境下,通过对cAMP-PKA信号通路抑制和增强的双向调节作用,以实现对以上寒、热两种证型的双向适用。

Objective：To study the regulation role of with neutral property to cAMP-PKA pathway in the rats with cold and heat blood stasis syndromes and it＇s mechanism. Methods ：60 rats were randomly divided into normal control group, model control group ,Se-men Persicae group, radix salvia mihiorrhiza group, rhizoma chuanxiong group, 12 rats per group. The three herb groups were orally giv-en relative herbs decoction, whose dosages were equal to 10 times the human clinical dose, normal and model control groups were orally given water,2 Limes/day ,20 mL/kg, for 7 days. Experiments in rats with cold and heat blood stasis syndromes were carried on respec-tiverly. In heat blood stasis syndromes, except normal control group, the other groups were intraperitoneally injected 10% carrageenan, 5 mL/kg, 1 Limes/day,for 3 days;24 hours after the last injection, subcutaneously injected 20% dry yeast suspension, 10 mL/kg. In cold blood stasis syndromes ,except normal control group, the other groups were put into fridge , temperature - （18 ± 2）℃ ,2 hours/ times,2 times/day,for 7 days. Separately draw 5ml abdominal aortic blood and taken abdominal aorta,6 hours and 12 hours after fin-ishing the model in the two syndromes. Tested the cAMP content by elisa,tested the PKA protein expression by Western blot. Results： Semen Persicae with neutral property, could decrease the content of cAMP in plasma（ P 〈 0.01 ） ,inhibit the expression of protein PKA （P 〈 O. 05 ）in rats with heat stagnation and blood stasis syndrome, increase the plasma content of cAMP（ P 〈 0.01 ） and increase the expression of protein PKA （P 〈 0.01 ）in rats with cold stagnation and blood stasis syndrome. Semen Persicae had two-way adjustment action on CAMP-PKA signal pathway. Conclusion：In different internal environment of heat stagnation and blood stasis syndrome, cold stagnation and blood stasis syndrome,Semen Persicae with neutral property has two-way adjustment to cAMP-PKA signaling channel, which may be one of the mechanism of it＇s two-way application.