Rat Anti-Mouse TLR2 monoclonal antibody, clone 6C2 Functional Grade

Astragaloside IV (AS/IV) is one of the extracted components from the traditional Chinese medicine Astragalus which has been demonstrated to have potential capacity for anti-inflammation activity and for treating cardiovascular disease. Our purpose was to determine the function and underlying molecular mechanism of AS/IV in hypoxia/reoxygenation (H/R) injured in cardiomyocytes. Differentially expressed genes (DEGs) were screened using bioinformatic analysis, and the molecular targeting relationship was verified by the dual-luciferase report system. H/R injured cardiomyocytes were employed to explore the effect of AS/IV. QRT-PCR and Western blot analysis were applied to detect the expression of mRNA and proteins, respectively. Additionally, superoxide dismutase (SOD), lactic dehydrogenase (LDH) and MDA (malondialdehyde) levels were detected to determine the oxidative damage. Cell viability was assessed by CCK-8, and flow cytometry was used to evaluate cell apoptosis ratio. TGFBR1 and TLR2 were selected as DEGs. Additionally, AS/IV could enhance cell proliferation and upregulated miR-101a expression, which suppressed TGFBR1 and TLR2 expression in H/R injured cardiomyocytes. Moreover, the results of Western blot exhibited that the downstream genes (p-ERK and p-p38) in the MAPK signaling pathway were suppressed, which meant AS/IV could inhibit this pathway in H/R injured cardiomyocytes. Overall, this study demonstrated AS/IV could attenuate H/R injury in human cardiomyocytes via the miR-101a/TGFBR1/TLR2/MAPK signaling pathway axis, which means that it could serve as a possible alternate for H/R treatment.

TLRs are cell-surface receptors which recognize a broad class of pathogen-associated molecular patterns (PAMPs) that activates innate and adaptive immune responses. Thorough perusal of literature suggests a possible role of TLR2 gene in resistance to infection due to gram positive bacteria. Bovine TLR2 assigns to Bos taurus (BTA) chromosome 17 it spans over 13.2 kb of genomic DNA which includes 2 exons. The first exon is spread over 179bp while the second one is 3333bp. Molecular characterization of TLR2 gene, carried out by PCR- SSCP analysis, revealed presence of six SNPs viz., A827G (exon 2.2), C1088G (exon 2.3), C2155A (exon 2.5), G2281C (exon 2.5), G2410A (exon 2.6) and C2600T (exon 2.6). Gene and genotypic frequencies were determined by values for allele frequencies and hetrozygosity. The degree of heterozygosities, chi(2) (Chi-square) test for Hardy Weinberg (HW) equilibrium and F statistics were performed for the HF crossbred population. Population genetic analysis demonstrated high degree of genetic differentiation in the population. The studied population displayed a deviation from the HW equilibrium.