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Description: tanshinone I is a type IIA human recombinant sPLA2 and Rabbit Recombinant cPLA2 inhibitor with IC50 of 11 respectively μ M and 82 μ M。

Related Categories: signaling pathways > > metabolic enzymes /
proteases > > phospholipids
Research field > > cardiovascular disease
Natural Products > > Quinones

Target:IC50: 11 μM (sPLA2), 82 μM (cPLA2)[1].

In Vitro Study: tanshinone I inhibited the formation of PGE2 by LPS induced raw macrophages (IC50 = 38 μ M）。 When tanshinone I and LPS were added at the same time, the compound significantly inhibited 10-100 μ PGE2 production of M (IC50 = 38 μ M）。 When added after complete induction of COX-2, tanshinone I also reduced the production of PGE2 (IC50 = 46) μ M）。 The fact that tanshinone I inhibits PGE 2 production through pre induced COX-2 strongly suggests that the compound can directly inhibit COX-2 activity and / or affect PLA2 Activity. When tanshinone I was incubated with two different forms of phospholipase A2 (PLA2), it significantly inhibited sPLA2 in a concentration dependent manner (IC50 = 11) μ M). Despite its low potency, tanshinone I also inhibited cPLA2 (IC50 = 82) μ M）[1]

In Vivo Study: tanshinone I showed anti-inflammatory activity in carrageenan induced paw edema and adjuvant induced arthritis in rats. In order to establish the anti-inflammatory activity of tanshinone I, classical animal models of acute and chronic inflammation [rat carrageenan (CGN) - induced paw edema and rat adjuvant induced arthritis (AIA)] were used. When oral tanshinone I, it showed significant anti-inflammatory activity against CGN induced paw edema (47% inhibition at 160 mg / kg), while the IC50 of indomethacin was 7.1 mg / kg. In AIA, tanshinone I gave 27% inhibition of secondary inflammation on day 18 at an oral dose of 50 mg / kg / day, while prednisolone (5 mg / kg / day) showed effective inhibition (65%) [1]

Kinase Experiment: as the source of PLA2, human recombinant sPLA2 (type IIA) was purified from CHO cells transfected with PLA2 gene, and rabbit recombinant platelet cPLA2 was obtained through its expression in baculovirus. Standard reaction mixture (200) μ 50) It contained 100 mm Tris HCl buffer (pH 9.0), 6 mM CaCl2 and 20 nmol of 1-acyl - [1-14C] - arachidonyl Sn glycerol phosphate ethanolamine (2000 CPM / nmol). Or there was no tanshinone I. the reaction was started by adding 50NG purified sPLA2 or cPLA2. After 20 minutes at 37 ℃, the free fatty acids produced were analyzed. Under these standard conditions, about 10% of free fatty acids are released from the added phospholipid substrate in the reaction mixture without tanshinone I [1].

Cell Experiment: raw 264.7 cells were cultured with DMEM supplemented with 10% FBS and 1% antibiotics at 37 ℃ under 5% CO2. In short, cells were seeded on 96 well plates (2) × 10 (5 cells / well). Unless otherwise indicated, LPS (1ug / ml) and tanshinone I were added and incubated for 24 hours. The PGE2 concentration in the medium was measured using the EIA kit for PGE2. In order to determine the effect of tanshinone I on PGE 2 production after COX-2 induction, cells were mixed with LPS (1 μ G / ml) for 24 hours and washed thoroughly. Then, tanshinone I was added without LPS and the cells were cultured for another 24 hours. PGE2 concentration was measured from the medium. MTT assay was used to examine the cytotoxicity of tanshinone I on raw cells. Tanshinone I at 100 μ M did not show any cytotoxicity [1].

Animal Experiment: in order to evaluate the inhibitory activity of tanshinone I on acute and chronic inflammatory animal models, rat carrageenan (CGN) - induced paw edema and adjuvant induced arthritis (AIA) models were used. In short, 1% CGN dissolved in pyrogen free saline (0.05 ml) was injected into the right rear paw of rats for paw edema test. After 5 hours, the swelling of the treated claws was measured using a plethysmograph. Tanshinone I dissolved in 0.5% CMC was administered orally 1 hour before CGN injection. For the AIA test, arthritic inflammation was induced by injecting Mycobacterium lactis (0.6ml / rat) dissolved in mineral oil into the right hind paw of rats. Tanshinone I was administered orally every day. The expansion of treated and untreated claws was measured using a plethysmograph.

References: [1]  Kim SY, et al. Effects of Tanshinone I isolated from Salvia miltiorrhiza bunge on arachidonic acid metabolism and in vivo inflammatory responses. Phytother Res. 2002 Nov; 16(7):616-20.