Rat liver tissue gene and protein expression in TLR4/NF-κB pathway.

<p><b>(A) Gene expression clustering tree and the corresponding standardized signal value of major expressed genes in the TLR4/NF-κB pathway</b>. Total RNA from 3 liver tissues (1 from the control group, 1 from the model group, and 1 from the TSG.M group) were harvested, amplified and labeled using an Agilent Whole Rat Genome Oligo Microarray (4 × 44 K) using Agilent SureHyb hybridization chambers. The processed slides were scanned with an Agilent microarray scanner G2505C after.hybridization and washing. Agilent Feature Extraction software was used to analyze acquired array images. Quantile normalization and subsequent data processing were performed with using the GeneSpring GX v11.5.1 software package. After quantile normalization of the raw data, genes that at least 1 out of 3 samples have flags in Detected (“All Targets Value”) were chosen for further data analysis. Differentially expressed genes between the two samples were identified through Fold Change filtering. Hierarchical Clustering was performed using the R scripts. <b>(B) Concentrations of FetA, TLR4, NF-κB, TNF-α, IL-1α, IL-6, and IL-10 proteins in the liver.</b> Tissue samples from the liver were excised and weighed after washing with 0.9% saline after rats were sacrificed using an intraperitoneal injection of 7% chloral hydrate (0.3 mL/100 g).100 mg tissues were rinsed with PBS and homogenized in 1 mL of PBS and then stored overnight at -20°C. Two freeze-thaw cycles were performed to break all cell membranes, and the homogenates were then centrifuged for 10 minutes at 4000 rpm, 4°C. The supernatant was collected for analysis. Concentrations of FetA, TLR4, NF-κB, TNF-α, IL-1α, IL-6, and IL-10 were tested using ELISA assay kits (mean±SD, n = 7). Statistical significance: * p< 0.05 vs. control;** p< 0.01 vs. control;*** p< 0.001 vs. control; ^# p< 0.05 vs. model; ^## p< 0.01 vs. model; ^### p< 0.001 vs. model