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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.
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.
Puerarin and icariin were detected in plasma and hippocampus after the oral administration，including plasma puerarin t1/2 2.45 h，icariin t1/2 3.59 h，hippocampus puerarin t1/2 4.37 h and icariin t1/2 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.
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.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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