Effects of Muscone on the Expression of P-gp, MMP-9 on Blood–Brain Barrier Model In Vitro

Abstract

Muscone is the main chemical ingredient in Musk which is main crude drug in Tongqiaohuoxue decoction (TQHXD), and TQHXD has a protective effect on damaged neurons, so we hypothesize that muscone can alter blood-brain barrier (BBB) permeability via the modulation of P-glycoprotein (P-gp) and matrix metalloproteinase-9 (MMP-9) expression. In this study, astrocytes (AC) and human umbilical vein endothelial cells (ECV304) were co-cultured to simulate the BBB model in vitro. Leak testing, transmembrane resistance experiments, and BBB-specific enzyme testing were used to test whether the model was successful. Different concentrations of muscone permeating the BBB were detected by gas chromatography (GC). The change of the transendothelial electrical resistance (TEER) on the BBB in vitro after treating with muscone was detected by Millicell-ERS. The protein expression of P-gp, MMP-9 in normal, and oxygen/glucose deprivation (OGD) BBB model was determined by western blotting to inquire that the mechanism of muscone penetrates the BBB model in vitro. The results show that muscone was detected in the lower medium of the BBB model by GC; the values of TEER were no significant difference before and after muscone (8 μM) was added to the BBB model; the expression of P-gp significantly decreased after the BBB model treatment with muscone (4, 8, and 16 μM) for 24 h; the expression of P-gp and MMP-9 in different concentrations of muscone groups had different degrees of reduction compared with the BBB in the state of OGD. In conclusion, muscone could permeate the BBB model, and it was associated with the inhibition of P-gp and MMP-9 expression. An understanding of the mechanisms of muscone across the BBB is crucial to the development of therapeutic modalities for cerebral vascular diseases.

Protection of Tong-Qiao-Huo-Xue Decoction against Cerebral Ischemic Injury through Reduction Blood–Brain Barrier Permeability

“…We have done some pharmacological experiments on these compounds. 34,35) These findings will increase our understanding of the effective substances of TQHXD. But the shortcoming of this paper is we do not isolate these compounds and prove their effects in comparison with the decoction.…”

In vitro acute and developmental neurotoxicity screening: an overview of cellular platforms and high-throughput technical possibilities

“…Attempts to fabricate the solid support for BBB models (Prieto et al 2004) include the use of transwell inserts (Wang et al 2015;Hind et al 2015), hollow fibres made of thermoplastic polymers (Cucullo et al 2011b), 3D printing (Kim et al 2015), and photolithography (Arayanarakool et al 2015). Creating in vitro models that mimic all BBB functions is particularly difficult, as endothelial cells (ECs) require exposure to shear stress to undergo the differentiation required for a well-functioning BBB (Cucullo et al 2011a).…”

Neuroprotective Effect Of Musk On Rat Brain Ischemic Model

“…Muscone can alter the permeability of BBB model in vitro, which is related to reduce the expression of P-gp and MMP-9. 34 Investigating the musk effects in protecting nerve cells from apoptosis in cerebral ischemia/reperfusion model of rat, the results have provided great opportunity to use musk in stroke treatment.…”