Abstract
BackgroundThe dry root and rhizome of Salvia miltiorrhiza Bunge, or Danshen, is a well-known traditional Chinese medicine with anticoagulant activity. Taking into account that thrombin (THR) and factor Xa (FXa) play crucial roles in the coagulation cascade, it is reasonable and meaningful to screening THR and/or FXa inhibitors from Danshen.MethodsFour extracts [butanol (BA), ethyl acetate (EA) and remained extract (RE) from 75% ethanol extract, and water extract (WE)] of Danshen were prepared, and their THR/FXa inhibitory activities were assessed in vitro. Then, the active EA extract was further separated by silica-gel column chromatography (SC), and its fractions (SC1–SC5) were analyzed by LC–MS. The principal component analysis (PCA) and orthogonal partial least squares discriminate analysis (OPLS-DA) were employed for predicting the specific marker compounds. The chemical structures of targeted compounds were identified by LC–MS/MS and their interactions with THR/FXa were analyzed by the molecular docking analysis.ResultsDanshen EA extract showed strong activity against THR and FXa, and its fractions (SC1–SC5) exhibited obvious difference in inhibitory activity against these two enzymes. Furthermore, four marker compounds with potential THR/FXa inhibitory activity were screened by PCA and OPLS-DA, and were identified as cryptotanshinone, tanshinone I, dihydrotanshinone I and tanshinone IIA. The molecular docking study showed that all these four tanshinones can interact with some key amino acid residues of the THR/FXa active cavities, such as HIS57 and SER195, which were considered to be promising candidates targeting THR and/or FXa with low binding energy (< − 7 kcal mol−1).ConclusionsLC–MS combined with multivariate statistical analysis can effectively screen potential THR/FXa inhibitory components in Danshen.
Highlights
The dry root and rhizome of Salvia miltiorrhiza Bunge, or Danshen, is a well-known traditional Chinese medicine with anticoagulant activity
Bioactivity‐guided fractionation The inhibitory activities against THR/factor Xa (FXa) of the ethyl acetate (EA), BA, remained extract (RE) and water extract (WE) extracts of Danshen (1.5 mg mL−1) and each positive control, argatroban and rivaroxaban (0.5 mg mL−1) were assessed, respectively
The results indicated that Danshen has the same effective components toward THR and FXa, which might be the similarity of chemical difference among the activity-based grouping [between high polar components (SC4 and SC5) and low polar components (SC1–SC3)]
Summary
The dry root and rhizome of Salvia miltiorrhiza Bunge, or Danshen, is a well-known traditional Chinese medicine with anticoagulant activity. Radix Salvia Miltiorrhiza, the dry root or rhizome of Saliva miltiorrhiza Bunge, namely Danshen in Mandarin, has been used to activate blood circulation to remove blood stasis in traditional Chinese medicine for more than thousands of years It is widely cultivated in China, such as in Shandong, Sichuan, Henan and Shaanxi provinces [1]. The modern pharmacological research showed that Danshen possesses multifarious pharmacological effects such as anticancer [3, 4], antiinflammatory [5], neuroprotection [6], anti-hypertension [7] and alleviation of diabetic retinopathy [8], etc It is one of the most widely applied Chinese medicines in the treatment of cardiovascular and cerebrovascular diseases [9, 10]. There are few studies reported about the thrombin (THR) or factor Xa (FXa) inhibitory activity of its extracts or ingredients
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