基于热敏感蛋白TRPV1的温热药性表达研究

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目的:探讨温热药对虚寒动物的热敏感蛋白(transient receptor potential vanilloid type-1,TRPV1)及体内cAMP(Cyclic Adenosine monophosphate)/PKA(protein kinase A)信号通路影响。方法:建立大鼠虚寒模型,应用温热中药黑顺片(30 g生药/kg)、干姜(20 g生药/kg)、肉桂(10 g生药/kg)及仙茅(30 g生药/kg)进行干预,应用放免法检测血清促甲状腺素(thyroid stimulating hormone,TSH)、甲状腺素(T4)、三碘甲状腺原氨酸(T3)、皮质酮(Corticosterone,COR)、环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP),应用免疫组化法检测大鼠下丘脑、海马、背根神经节中热敏感蛋白(TRPV1)、磷酸化蛋白激酶A(Phospho-(Ser Thr)PKA,PPKA)蛋白表达。结果:虚寒组大鼠出现活动减退、爱扎堆、毛色枯萎等现象,血清中TSH、T3、T4、COR、cAMP、c GMP含量降低,大鼠下丘脑、海马、背根神经节中TRPV1、PPKA蛋白表达明显降低,黑顺片(30 g生药/kg kg)、干姜(20 g生药/kg)、肉桂(10 g生药/kg)及仙茅(30g生药/kg)干预后,大鼠活动性增加,毛色改善,血清中TSH、T3、T4、COR、cAMP、c GMP含量明显升高,大鼠下丘脑、海马、背根神经节中TRPV1、PPKA表达明显增强。结论:热性药可改善虚寒状态内分泌水平,其作用机制可能与增强虚寒机体时下调的TRPV1、PPKA蛋白表达有关,从而表达出温热药性。 OBJECTIVE: To investigate the effect of warming medicine on heat-responsive protein (TRPV1) and cAMP (Cyclic Adenosine monophosphate) / PKA (protein kinase A) signaling pathway in cold-blooded animals. Methods: The rat model of Deficiency of Asthenia was established. The effects of warm traditional Chinese medicine Heshun Tablet (30 g crude drug / kg), ginger (20 g crude drug / kg), cinnamon (10 g crude drug / kg) kg). Serum levels of thyroid stimulating hormone (TSH), thyroxine (T4), triiodothyronine (T3), corticosterone (COR) and cAMP cAMP and cGMP, and the expressions of TRPV1, Phospho- (Ser Thr) PKA, cTnA and cGMP in rat hypothalamus, hippocampus and dorsal root ganglion were detected by immunohistochemistry. PPKA) protein expression. Results: The activities of TSH, T3, T4, COR, cAMP and c GMP in serum were decreased in the rats of Deficiency group, the activities of TRPV1 in the hypothalamus, hippocampus and dorsal root ganglion were decreased, The expression of PPKA protein was significantly decreased in the rats treated with hematoxylin and eosin (30 g crude drug / kg kg), ginger (20 g crude drug / kg), cinnamon (10 g crude drug / kg) Activity increased, fur color improved, serum TSH, T3, T4, COR, cAMP, c GMP content was significantly increased in rats hypothalamus, hippocampus and dorsal root ganglion TRPV1, PPKA expression was significantly enhanced. CONCLUSION: The thermo-medicine can improve the endocrine level of Deficiency-Cold state, and its mechanism may be related to the down-regulation of TRPV1 and PPKA protein expression in reducing the body temperature so as to express the thermo-medicinal properties.
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