Pr6502

The total RNA was extracted from the liver and using TRIpure regent (RP1001, BioTeke). The cDNA was synthesized from RNA by a reverse transcription reagent kit (PR6502, BioTeke). The expressions of these genes (shown in Table 1) were determined by qRT-PCR and using SYBR Green PCR kit (SY1020, Solarbio), which were performed on Real-Time PCR System (ExicyclerTM 96, BIONEER, Korea). ß-Actin was used as a reference gene and comparison in expression between groups was made using the 2^−ΔΔCT method.

Expression of proinflammatory cytokines TNF-α, IL-1β, IL-6, and IFN-γ in kidney tissue was detected using qRT-PCR. Briefly, total RNA was extracted from homogenized kidney tissue using a TRI pure Reagent assay kit (RP1001, BioTeke, BJ, CHN) according to the manufacturer's instructions. The RNAs from each group were reverse-transcribed using a reverse transcription kit (PR6502, BioTeke, BJ, CHN) to obtain the corresponding cDNA. The analysis of gene expression was performed in the Exicycler 96 Real Time PCR System (BIONEER, Daejeon, ROK) using a SYBR Green kit (SY1020, Solarbio, BJ, CHN). The changes in mRNA were normalized to the control (β-actin). Each sample was tested at least three times, and the average value was taken. The sequences of primers used for qRT-PCR are listed in Table 1.