熟地黄对阴虚模型大鼠血清褪黑素及脑内γ-氨基丁酸的影响

Effect of Rehmanniae Radix Praeparata on Serum Melatonin and GABA in Yin Deficiency Rats

目的观察熟地黄对阴虚模型大鼠血清褪黑素及脑内γ-氨基丁酸(GABA)的影响,探讨其滋阴宁静的作用机制。方法将雄性SD大鼠随机分为正常组、模型组、熟地黄组(30 g·kg^(-1)),每组10只;模型组、熟地黄组灌胃耗阴复方药液(30 g·kg^(-1)),每日1次,连续10 d;第11天熟地黄组开始灌胃熟地黄药液,每日1次,连续10 d。观察大鼠的一般情况,测定体质量、饮水量,通过旷场行为学检测其活跃度;采用高通量测序检测大鼠脑组织中阴虚相关基因的差异表达;ELISA法检测大鼠血清褪黑素含量;高效液相色谱法检测大鼠脑组织中GABA含量;Western Blot法检测下丘脑组织中GABA合成酶Gad67及其转录因子EGR1的蛋白表达。结果与正常组比较,模型组大鼠的体质量明显下降(P<0.05),饮水量以及活跃度明显提高(P<0.05);促甲状腺激素释放激素(TRH)及促肾上腺皮质激素释放激素(CRH)基因表达量明显提高(P<0.05);血清褪黑素含量、脑组织GABA含量,以及Gad67和EGR1的蛋白表达量均明显下降(P<0.05,P<0.01)。与模型组比较,熟地黄组大鼠体质量明显升高(P<0.05),饮水量以及活跃度明显下降(P<0.05);TRH及CRH基因表达量明显降低(P<0.05,P<0.01);血清褪黑素含量、脑组织GABA含量,以及Gad67和EGR1的蛋白表达量均明显升高(P<0.05,P<0.01)。结论熟地黄可能通过提高大鼠血清褪黑素及脑内GABA水平来发挥其滋阴宁静的作用。

Objective To study the tranquilizing effect of Rehmanniae radix praeparata on yin deficiency model rats and its effect on the expression of melatonin and GABA. Methods Male SD rats were randomly divided into normal group,model group,and Rehmanniae radix praeparata(Shudi)group(30 g·kg^(-1)),with 10 rats in each group. Rats in the model group and the Shudi group were given the compound medicine solution(30 g·kg^(-1))once every day for10 days to establish the yin-deficiency model. On the 11 th day,the Shudi group rats were given the Rehmannia decoction for 10 consecutive days. The normal group was given the same volume of distilled water for 20 days. By observing the general situation of the animals, water intake and the activity of open field behavior, whether the model was successfully developed was determined. High-throughput sequencing was used to detect the differential expression of yin deficiency related genes in rats’ brain tissue. ELISA kit was used to detect the serum content of melatonin. High performance liquid chromatography was used to detect the content of GABA in rat’s brain tissue.Western Blot was used to detect the expression of yin deficiency related proteins,Gad67 and EGR1,in rat’s brain tissue. Results Compared with the normal group,the body weight of the model group was decreased significantly(P<0.05);the water intake and activity were significantly increased(P<0.05);the gene expression of thyroid stimulating hormone releasing hormone(TRH) and adrenocorticotropic hormone releasing hormone(CRH) were significantly increased(P<0.05);the content of melatonin in in serum,the content of GABA in brain tissue,and the protein expression of Gad67 and EGR1 were significantly decreased in the model group(P<0.05,P<0.01).Compared with the model group, the body weight of the shudi group was increased significantly(P<0.05);the amount of drinking water and activity(P<0.05),TRH and CRH gene expression were decreased(P<0.05,P<0.01);serum melatonin content,brain GABA content,and Gad67 and EGR1 protein expression were significantly increased in the Shudi group(P<0.05, P<0.01). Conclusion Rehmanniae radix praeparata can improve the contents of serum melatonin and brain GABA,and play the role of nourishing yin and tranquility.