Identification of potential regulating effect of baicalin on NFκB/CCL2/CCR2 signaling pathway in rats with cerebral ischemia by antibody-based array and bioinformatics analysis

Abstract

Ethnopharmacological relevanceBaicalin is one of the major bioactive compounds extracted from the root of Scutellaria baicalensis Georgi, which was used to treat cerebral ischemia for thounds of years. However, its biological mechanisms remains to be further explored. Aim of the reviewThis study aims to identify potential biological mechanisms of baicalin against cerebral ischemia combining antibody-based array and bioinformatics analysis. MethodsA rat model of middle cerebral artery occlusion (MCAO) was constructed. Sprague-Dawley rats were randomly divided into three groups: control group, ischemic model group, and baicalin 100 mg/kg treatment group respectively. Bederson score and 2, 3, 5-triphenyl tetrazolium chloride (TTC) staining were examined to evaluate the pharmacodynamics of baicalin treatment. Antibody-based array technology, enzyme linked immunosorbent assay (ELISA), western-blot, molecular docking, transcription factor perdiction, hematoxylin and eosin (H&E), and immunofluorescence staining were used to study the regulation of baicalin on inflammatory response after cerebral ischemia in vivo. LPS-induced RAW 264.7 macrophage inflammation model was prepared to observe the anti-inflammatory effect of baicalin in vitro. ResultsBaicalin (100 mg/kg) reduced neurological injury score, cerebral infarction volume, and necrotic cells in MCAO rats. Baicalin inhibited the expression of CCL2, and reduced the phosphorylation levels of p65, IκBα protein and down-regulated level of CCR2. Besides, baicalin could bond to CCR2 directly, which prevented CCL2 from binding to CCR2. Furthermore, baicalin down-regulated the number of monocytes in the peripheral blood and improved the spleen index post-cerebral ischemia. In vitro, baicalin significantly inhibited the secretion of NO, IL6, TNFα, and CCL2 in macrophages and promoted the secretion of IL13, IFNG, and IL1a. ConclusionsBaicalin inhibited cerebral ischemia-induced activation of the NFκB/CCL2/CCR2 pathway with multiple target effect. These data promote the therapeutic utilization of baicalin in preventing cerebral ischemia clinically.