The mRNA levels of SUR1, TRPM4, Il-1, Il-10, TNFα, and GAPDH were routinely measured by quantitative real-time polymerase chain reaction (qPCR). Briefly, total RNA was isolated using a total RNA/DNA isolation kit (Tiangen, Peking, China) and reverse transcribed to cDNA with PrimeScriptTM RT Master Mix Kit (TAKARA) according to the manufacturer's instructions. qPCR was performed using the SYBR Green master mixes (TAKARA) and Roche LightCycler480 System. Relative changes of mRNA expression were normalized to the level of GAPDH.
The Tested genes and primer sequences were listed as follows:
Abcc8: Forward primer 5′-3’: CATCCGGGTGAGGAGATACG;
Reverse primer 5′-3’: CAGGTTAACGAAGGGCTGCA.
Trpm4: Forward primer 5′-3’: TGATGAGCACACCACGGAGA;
Reverse primer 5′-3’: ATCCGTGCGATCAGACAGC.
Il-1: Forward primer 5′-3’: CCTACTTCAGCATCCTCTACTGG;
Reverse primer 5′-3’: AGGGTTTCTTGAGAAGGGGAC.
Il-10：Forward primer 5′-3’: CCCATTCCTCGTCACGATCTC;
Reverse primer 5′-3’: TCAGACTGGTTTGGGATAGGTTT.
Tnfa: Forward primer 5′-3’: CACTCTGGGTACGTGGGTG;
Reverse primer 5′-3’: CACAGGTGATAATGAGGACAGC.
Ccnd1: Forward primer 5′-3’:GCGTACCCTGACACCAATCTC;
Reverse primer 5′-3’: CTCCTCTTCGCACTTCTGCTC.
Nlrp3: Forward primer 5′-3’: ATTACCCGCCCGAGAAAGG;
Reverse primer 5′-3’: TCGCAGCAAAGATCCACACAG.
Gapdh: Forward primer 5′-3’: AGGTCGGTGTGAACGGATTTG;
Reverse primer 5′-3’: TGTAGACCATGTAGTTGAGGTCA.
The Tested genes and primer sequences were listed as follows:
Abcc8: Forward primer 5′-3’: CATCCGGGTGAGGAGATACG;
Reverse primer 5′-3’: CAGGTTAACGAAGGGCTGCA.
Trpm4: Forward primer 5′-3’: TGATGAGCACACCACGGAGA;
Reverse primer 5′-3’: ATCCGTGCGATCAGACAGC.
Il-1: Forward primer 5′-3’: CCTACTTCAGCATCCTCTACTGG;
Reverse primer 5′-3’: AGGGTTTCTTGAGAAGGGGAC.
Il-10：Forward primer 5′-3’: CCCATTCCTCGTCACGATCTC;
Reverse primer 5′-3’: TCAGACTGGTTTGGGATAGGTTT.
Tnfa: Forward primer 5′-3’: CACTCTGGGTACGTGGGTG;
Reverse primer 5′-3’: CACAGGTGATAATGAGGACAGC.
Ccnd1: Forward primer 5′-3’:GCGTACCCTGACACCAATCTC;
Reverse primer 5′-3’: CTCCTCTTCGCACTTCTGCTC.
Nlrp3: Forward primer 5′-3’: ATTACCCGCCCGAGAAAGG;
Reverse primer 5′-3’: TCGCAGCAAAGATCCACACAG.
Gapdh: Forward primer 5′-3’: AGGTCGGTGTGAACGGATTTG;
Reverse primer 5′-3’: TGTAGACCATGTAGTTGAGGTCA.