参葛补肾胶囊中葛根素和淫羊藿苷小鼠体内动力学脑海马分布及其相关蛋白动态表达的相关性研究*

王卫华; 雷帆; 李成功; 孙虹; 胡时先; 邢东明; 任宾; 郝娟; 杜力军

doi:10.16155/j.0254-1793.2024-0423

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药物分析杂志 ›› 2024, Vol. 44 ›› Issue (11) : 1909-1922. DOI: 10.16155/j.0254-1793.2024-0423

生物检定·代谢分析

参葛补肾胶囊中葛根素和淫羊藿苷小鼠体内动力学脑海马分布及其相关蛋白动态表达的相关性研究^*

王卫华^1,2, 雷帆¹, 李成功³, 孙虹⁴, 胡时先³, 邢东明¹, 任宾³, 郝娟³, 杜力军^1**

作者信息 +

Study on the pharmacokinetics and the correlation between distribution of puerarin-icariin and dynamic expression of proteins in mouse hippocampus after oral administration of Shenge Bushen capsules^*

WANG Wei-hua^1,2, LEI Fan¹, LI Cheng-gong³, SUN Hong⁴, HU Shi-xian³, XING Dong-ming¹, REN Bin³, HAO Juan³, DU Li-jun^1**

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摘要

目的:研究参葛补肾胶囊中葛根素和淫羊藿苷在小鼠血浆及脑内海马核团动力学和海马抑郁症相关蛋白表达的变化,探讨参葛补肾胶囊体内药代动力学变化及其与海马相关蛋白表达的相关性。方法:运用UPLC-MS/MS-ABSCIEX QTRAP 5500三重四极杆串联线性离子阱质谱方法,采用Waters ACQUITY HSS T3(50 mm×2.1 mm, 1.8 μm)色谱柱,以0.05%甲酸水-含0.05%甲酸的乙腈甲醇溶液(1∶1)为流动相梯度洗脱,流速0.2 mL·min^-1,采用电喷雾离子源,在负离子模式下对正常小鼠灌胃后不同时间点的血浆及海马中葛根素、淫羊藿苷测试分析。用Western blot方法表达海马蛋白。结果:口服给药后血浆和海马均检测到葛根素和淫羊藿苷,其中血浆葛根素半衰期(t_1/2)为2.45 h, 淫羊藿苷t_1/2为3.59 h;海马葛根素t_1/2为4.37 h,淫羊藿苷t_1/2为8.5 h。葛根素血浆浓度占给药量的27.3%,淫羊藿苷血浆浓度占给药量的1.34%。海马葛根素占吸收入血葛根素的2.47%,海马淫羊藿苷占吸收入血淫羊藿苷的73.56%。给药后海马神经元限制性沉默因子(NRSF)、脑源性神经营养因子(BDNF)及其下游原肌球蛋白相关激酶受体B(TrkB)、溶质载体转运体6a4(SLC6A4)、糖皮质激素受体(GR)、μ阿片受体(MOR)等蛋白表达出现不同程度的上调,其中淫羊藿苷与NRSF表达呈现明显负相关,葛根素与BDNF-TrkB、MOR表现出明显正相关,GR则无明显相关性。结论:小鼠口服参葛补肾胶囊后,葛根素和淫羊藿苷均能进吸收进入血液且分布于海马,淫羊藿苷较葛根素更易透过血脑屏障并在海马分布。海马相关蛋白表达与葛根素、淫羊藿苷浓度变化呈现一定的相关性,提示2个成分的作用靶点与这些蛋白有关。本研究为参葛补肾胶囊药代动力学及其抗抑郁作用的物质基础提供了重要的实验依据。

Abstract

Objective: To study the dynamic changes of puerarin and icariin in plasma and hippocampus of mice, to observe the changes of depression-related protein expression in hippocampus, and to explore the pharmacokinetic changes of Shenge Bushen capsules (SBC) and its correlation with the changes of hippocampus-related protein expression. Methods: Puerarin and icariin in plasma and hippocampus of normal mice were analyzed by UPLC-MS/MS-ABSCIEX QTRAP 5500 triple quadrupole series linear ion trap mass spectrometry method using Waters Acquity HSS T3 (50 mm×2.1 mm, 1.8 μm) column with 0.05% formic acid water-acetonitrile-methanol (1∶1) containing 0.05% formic acid as mobile phases gradient elution at a flow rate of 0.2 mL·min^-1 and electrospray ion source under negative ion model. The proteins in hippocampus were expressed by Western blot assay. Results: Puerarin and icariin were detected in plasma and hippocampus after the oral administration, including plasma puerarin t_1/2 2.45 h, icariin t_1/2 3.59 h, hippocampus puerarin t_1/2 4.37 h and icariin t_1/2 8.5 h. The plasma concentration of puerarin accounts for 27.3% and that of icariin accounts for 1.34% of the dosage taken. Puerarin in hippocampus accounts for 2.47% of puerarin absorbed into blood, while icariin in hippocampus accounts for 73.56% of icariin absorbed into blood. The expressions of restrictive silencing factor (NRSF), brain-derived neurotrophic factor (BDNF) and its downstream protein tyrosine kinase B (TrkB), solute carrier transporter 6a4 (SLC6A4), glucocorticoid receptor (GR) and μ opioid receptor (MOR) in hippocampus were all up-regulated after SBC administration, in which icariin was negatively correlated with NRSF expression, puerarin was positively correlated with BDNF-TrkB and MOR, and GR had no obvious correlation with the proteins. Conclusions: Puerarin and icariin can be absorbed into the blood and distributed in the hippocampus after oral administration of SBC in mice. Icariin was easier to pass through the blood-brain barrier and distribute in the hippocampus than puerarin. The expression of hippocampus-related proteins was related to the changes of puerarin and icariin concentrations, suggesting that the targets of the two components as well as SBC were related to these proteins. This study provides an important experimental basis for the pharmacokinetic study of SBC and also provides a material basis of its antidepressant effect.

导出引用

王卫华, 雷帆, 李成功, 孙虹, 胡时先, 邢东明, 任宾, 郝娟, 杜力军. 参葛补肾胶囊中葛根素和淫羊藿苷小鼠体内动力学脑海马分布及其相关蛋白动态表达的相关性研究^*[J]. 药物分析杂志, 2024, 44(11): 1909-1922 https://doi.org/10.16155/j.0254-1793.2024-0423

WANG Wei-hua, LEI Fan, LI Cheng-gong, SUN Hong, HU Shi-xian, XING Dong-ming, REN Bin, HAO Juan, DU Li-jun. Study on the pharmacokinetics and the correlation between distribution of puerarin-icariin and dynamic expression of proteins in mouse hippocampus after oral administration of Shenge Bushen capsules^*[J]. Chinese Journal of Pharmaceutical Analysis, 2024, 44(11): 1909-1922 https://doi.org/10.16155/j.0254-1793.2024-0423

中图分类号： R 917

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