Angelica sinensis polysaccharide attenuates constipation-associated colorectal carcinogenesis by inhibiting the NLRP3/Caspase-1/IL-1β inflammasome axis

Table 1 Oligonucleotide primer sequences used for reverse-transcription PCR analysis of inflammatory mediators

Gene	Forward primer (5′-3′)	Reverse primer (5′-3′)	Product size (bp)
IL-1β	5′-ATGATGGCTTATTACAGTGGCAA-3′	5′-GTCGGAGATTCGTAGCTGGA-3′	132
IL-18	5′-TCTTCATTGACCAAGGAAATCGG-3′	5′-TCCGGGGTGCATTATCTCTAC-3′	145
TNF-α	5′-CCTCTCTCTAATCAGCCCTCTG-3′	5′-GAGGACCTGGGAGTAGATGAG-3′	220
IL-6	5′-ACTCACCTCTTCAGAACGAATTG-3′	5′-CCATCTTTGGAAGGTTCAGGTTG-3′	149
COX-2	5′-CTGGCGCTCAGCCATACAG-3′	5′-CGCACTTATACTGGTCAAATCCC-3′	183
MMP-9	5′-TGTACCGCTATGGTTACACTCG-3′	5′-GGCAGGGACAGTTGCTTCT-3′	97
GAPDH	5′-GGAGCGAGATCCCTCCAAAAT-3′	5′-GGCTGTTGTCATACTTCTCATGG-3′	197

All primers were designed based on human mRNA sequences retrieved from the NCBI GenBank database and synthesized by Sangon Biotech (Shanghai, China). Primer specificity was confirmed by melting curve analysis and agarose gel electrophoresis. Glyceraldehyde-3-phosphate dehydrogenase served as the endogenous reference gene for normalization. GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; TNF-α: Tumor necrosis factor-alpha; MMP-9: Matrix metalloproteinase-9; IL-1β: Interleukin-1 beta; IL-18: Interleukin-18; IL-6: Interleukin-6.