Danshen extract (Salvia miltiorrhiza Bunge) attenuate spinal cord injury in a rat model: A metabolomic approach for the mechanism study.

BACKGROUD: Spinal cord injury (SCI) is a devastating neurological disorder caused by trauma. To date, SCI treatment is still a significant challenge in clinic and research around the world. Danshen (dried roots and rhizomes of Salvia miltiorrhiza), a commonly used Chinese medicinal herb, has been attracting attention in SCI treatment. PURPOSE: Aim of this study was to evaluate the potential beneficial effects of danshen extract in a SCI rat model, as well as investigate possible mechanism of action and potential biomarkers. METHODS: Here, a rat SCI model was established with weight-drop method, and danshen extract was administered by oral gavage (12.5 g/kg). Recovery of motor function and histomorphological changes were evaluated by Basso, Beattie and Bresnahan score and hematoxylin-eosin staining, respectively. In addition, neurofilament 200 (NF-H), brain-derived neurotrophic factor (BDNF), glial fibrillary acidic protein (GFAP) and CD11b expressions were assayed by immunofluorescence and western blot analysis. Furthermore, a metabolomics analysis based on ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) approach was conducted. RESULTS: The results demonstrated that danshen extract could significantly ameliorated histopathology changes and improved recovery of motor function after SCI. Moreover, NF-H, BDNF and CD11b expression were progressively increased until 4 weeks post-injury after administrated danshen extract. Furthermore, a good separation was observed among different groups using OPLS-DA. Trajectory analysis showed the gradual shift from position of model group toward normal group with increasing time after administration of danshen extract. Meanwhile, 51 significantly altered metabolites were identified, while metabolic pathway analysis suggested that 6 metabolic pathways were disturbed by the altered metabolites. CONCLUSION: In summary, this study provides an overview of neuroprotective effects and investigates possible mechanism of danshen extract in SCI treatment. However, further research is needed to uncover its regulatory mechanisms more clearly.

Comparative pharmacokinetic study on phenolic acids and flavonoids in spinal cord injury rats plasma by UPLC-MS/MS after single and combined oral administration of danshen and huangqin extract.

Chinese medicinal herbs danshen and huangqin have attracted attention in spinal cord injury (SCI) treatment. Purpose of this study was to investigate and compare the pharmacokinetic characteristics of 4 phenolic acids and 4 flavonoids in SCI rat plasma after orally administrate danshen, huangqin and combined extract of these two herbs (CDH). Thus, a rapid and sensitive ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for simultaneously quantitative determination of tanshinol, protocatechualdehyde, protocatechuic acid, salvianolic acid A, baicalein, baicalin, wogonin and wogonoside. After inducing a contusion injury by a weight-drop device, SCI rats were orally administrated a single dose (12.5 g/kg) of danshen, huangqin and CDH extracts, respectively. Then, blood samples at different time points were collected and analyzed. In CDH group, Cmax and AUC of tanshinol, protocatechualdehyde and protocatechuic acid significantly declined, while those of salvianolic acid A enhanced. These changes were beneficial for danshen to treat SCI. As for flavonoids, double peaks were observed in huangqin group, while this phenomenon disappeared in CDH group. Concomitantly, Cmax and AUC declined after administrated CDH. These alterations were due to influence of danshen active constituents on absorption and transportation process of flavonoids. Therefore, danshen and huangqin significantly influenced pharmacokinetic profile and parameters of each other, thus exert synergistic therapeutic effect in SCI treatment.