Simultaneous Determination of 17 Constituents of Chinese Wild Radix Salvia miltiorrhiza from Different Geographical Areas by Ultra-High Performance Liquid Chromatography Coupled to Triple Quadrupole Mass Spectrometry

Background and Objective: Radix Salvia miltiorrhiza (RSM) has been used clinically for the prevention and treatment of cardiovascular diseases; therefore, it is important to strengthen its quality management. Considering multiple constituents when assessing RSM quality is essential. We established a simple, rapid method to identify and quantify the major bioactive constituents in RSM using ultra high performance liquid chromatography (UPLC) coupled to a triple quadruple mass spectrometry (QqQ-MS). Methods: We analyzed 17 markers from 50 batches of wild S. miltiorrhiza samples that were collected from different locations in China. The ultrasonic extracts of all samples were determined using the UPLC-QqQ-MS method and were assessed by hierarchical cluster analysis (HCA). Results: We used this method to analyze 50 sample batches of the 17 compounds and obtained results with excellent linearity (R2, 0.9915-0.9997), precision (relative standard deviation, RSD, 0.15-1.94%), repeatability (RSD, 1.28-4.71%), stability (RSD, 0.97-5.60%) and recovery (RSD, 0.305-6.40%). The hierarchical cluster analysis was used to classify the 50 samples based on the characteristics of the 17 compound markers. Conclusion: We demonstrated that the developed method was simple, reproducible and sensitive, and it is capable of systematic and scientific evaluation for quality control of RSM. The HCA clearly demonstated that the RSM samples from different locations were significantly different and the quality of wild Radix S. miltiorrhiza could generally be judged according to its geographical origin.

Tanshinone IIA attenuates Aβ-induced neurotoxicity by down-regulating COX-2 expression and PGE2 synthesis via inactivation of NF-κB pathway in SH-SY5Y cells

Amyloid-β (Aβ)-induced neurotoxicity is a major pathological mechanism of Alzheimer’s disease (AD). Tanshinone IIA (Tan IIA), extracted from traditional Chinese herb Radix salvia miltiorrhiza, possesses anti-oxidant and anti-inflammatory actions, as well as neuroprotective effects. The present study aims to explore the possible mechanism by which Tan IIA attenuated Aβ-induced neurotoxicity. Exposure of SH-SY5Y cells to different concentrations of Aβ led to neurotoxicity by reducing cell viability, inducing cell apoptosis and increasing neuroinflammation in a dose-dependent manner. Moreover, Aβ treatment promoted cyclooxygenase-2 (COX-2) expression and Prostaglandin E2 (PGE2) secretion, and activated nuclear transcription factor kappa (NF-κB) pathway in SH-SY5Y cells. However, pretreatment of SH-SY5Y cells with Tan IIA prior to Aβ prevented these Aβ-induced cellular events noticeably. These data suggested that Tan IIA exerted its neuroprotective action by alleviating Aβ-induced increase in COX-2 expression and PGE2 secretion via inactivation of NF-κB pathway.

Electrospray Ionization Mass Spectral Characteristics and Fragmentation Mechanisms of Tanshinones in Radix Salvia miltiorrhiza

Cryptotanshinone, tanshinone I, and tanshinone IIA were analyzed by High-performance liquid chromatography /electrospray ionization-mass spectrometry. MSn spectra were obtained and optimized by energy collision induced dissociation (CID) from [M+H]+ ions, the effect of collision energy on production of fragmental ions were studied and optimal signals were achieved. With the collision energy of 35%, tanshinones have optimal signals of fragmental ions and maximal amounts of product fragment ions respectively. The fragmentation pathways for the compounds were studied, and this information would be helpful for the quantitative and pharmacokinetic analysis of tanshinones.

Salvianolic Acid B Inhibits ERK and p38 MAPK Signaling in TGF-β1-Stimulated Human Hepatic Stellate Cell Line (LX-2) via Distinct Pathways

Salvianolic acid B (SA-B) is water-soluble component ofRadix Salvia miltiorrhiza. The previous work indicated that SA-B can inhibit MAPK and Smad signaling in activated hepatic stellate cells (HSCs) to perform anti-fibrotic activity Lv et al. 2010. However, some studies have shown that there is cross-talk between MAPK and Smad in certain cell types. Thus, the anti-fibrotic action of SA-B may be through the cross-talk. In order to clarify the mechanism of SA-B further, we knocked down Smad in LX-2 cells (SRV4) via RNAi, and then added TGF-β1, and PD98059 or SB203580 and SA-B. The levels of p-MEK and p-p38 were inhibited by SA-B in SRV4 independent of TGF-β1. The expression of Col I andα-SMA in SRV4 could be reduced by SA-B independent TGF-β1. SB203580 had not significant effect on p-MEK in SRV4 stimulated by TGF-β1. The levels of p-MEK in SRV4 were not increased significantly after TGF-β1 stimulation. PD98059 had no effect on the levels of p-p38 in SRV4 irrespective of TGF-β1. In conclusion, SA-B inhibits the synthesis of Col I in LX-2 cells independent of TGF-β1 stimulation, and the anti-fibrotic effect of SA-B is due to direct inhibition of p38 signaling and inhibition the cross-talk of Smad to ERK signaling.