金匮肾气丸对肾阳虚体质大鼠单胺类神经递质水平的影响

Effect of Jingui Shenqi Pill on the Levels of Monoamine Neurotransmitters in Rats with Kidney Yang Deficiency Constitution

目的研究低温环境、寒凉饮食诱导的肾阳虚体质大鼠单胺类神经递质水平改变及金匮肾气丸的调控作用机制。方法大鼠肾阳虚体质模型以0.54 g/(kg·d)盐黄柏配方颗粒灌胃、1 h/d处于4℃人造寒冷环境并传代的方法制备,亲代造模持续至子代断乳,子代造模持续至其12周龄。亲代对照组所生为子代对照组,亲代模型组所生随机分为子代模型组和子代干预组。子代干预组自8周龄起给予1.08 g/(kg·d)金匮肾气丸干预。选取子代各组雄鼠为研究对象,每组6只。旷场实验、温度趋向性考察观察各组大鼠行为改变,ELISA法检测海马组织脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)、酪氨酸激酶受体B(tyrosine kinase receptor B,TrkB)、环磷腺苷效应元件结合蛋白(cyclic adenosine monophosphate response element-binding,CREB)、经典瞬时受体电位通道(transient receptor potential cation channel,TRPC)3、TRPC6及海马组织、血清中的5-羟色胺(5-hydroxytryptamine,5-HT)、多巴胺(dopamine,DA)、去甲肾上腺素(norepinephrine,NE)含量。结果与对照组比较,模型组大鼠旷场实验运动距离、运动速度显著降低(P<0.05),静止不动时间显著增加(P<0.05),温度趋向性考察热板停留时间以及海马与血清中上述各项指标水平均显著增加(P<0.05)。与模型组比较,干预组大鼠运动距离、运动速度显著增加(P<0.05),静止不动时间、热板停留时间以及上述各项指标水平均显著下降(P<0.05)。结论金匮肾气丸能够回调肾阳虚体质导致的单胺类神经递质水平异常升高,其机制可能与调节BDNF/TrkB/CREB通路和以TRPC3/6为代表的经典瞬时受体电位家族有关。

Objective To study the changes in monoamine neurotransmitter levels in rats with kidney Yang deficiency constitution induced by low-temperature and cold food diet,and the regulatory mechanism of Jingui Shenqi pill.Methods The model of kidney Yang deficiency constitution in rats was prepared by administering 0.54g/(kg·d)salted Cortex Phellodendri granules,exposure to an artificial cold environment of 4℃for 1 hour per day,and passage.The model was maintained until the offspring rats were weaned,and the modeling continued until they were 12 weeks old.The offspring of the control group of the parent rats were the control group of the offspring,and the offspring of the model group of the parent rats were randomly divided into the model group and the intervention group of the offspring.The intervention group of the offspring was given 1.08 g/(kg·d)Jingui Shenqi pill intervention from 8 weeks of age.Male rats from each group were selected as the research subjects,with 6 rats in each group.Their behavioral changes were observed using an open field experiment and temperature preference test,and the levels of BDNF,TrkB,CREB,TRPC3,TRPC6,5-HT,DA,and NE in the hippocampus tissue or serum were detected using ELISA method.Results Compared with the control group,the model group showed significantly decreased distance and speed of movement in the open field experiment(P<0.05),significantly increased immobility time(P<0.05),and significantly increased hot plate stay time in the temperature preference test(P<0.05),as well as significantly increased levels of various molecular indicators above in hippocampus tissue or serum(P<0.05).Compared with the model group,the intervention group showed significantly increased distance and speed of movement in the open field experiment(P<0.05),significantly decreased immobility time(P<0.05),significantly decreased hot plate stay time in the temperature preference test(P<0.05),as well as significantly decreased levels of various molecular indicators above(P<0.05).Conclusion Jingui Shenqi pill can regulate the abnormally elevated levels of monoamine neurotransmitters caused by kidney Yang deficiency constitution.Its mechanism may be related to the BDNF/TrkB/CREB pathway and the classic transient receptor potential channel family represented by TRPC3/6.