2.3 RT-PCR
After total mRNA of all the rabbits cartilage samples was extracted using TRIzol Kit and reversely transcribed, polymerase chain reactionwas manipulated to cDNA, with the SYBR Premix Ex Taq. Then, gene expression of mRNA was calculated using the cycle threshold method (2−ΔΔCt). The primer sequences were as follows:
TLR1 F CTTGATCTGCACAATAACCG R GAAGATCCACCAAGGAGT
TLR2 F TGCCCTGTTCATTTTCCC R TTGCCTTAAAATCAACGTGTG
TLR3 F TTTCAAGGGCTTAGCTCA R AAACATTCTTGTCAACGG
TLR4 F ACAAGGCTTACTTTCACT R TCTACGCTCAGAACAGCA
TLR5 F TCTGATTCTCGCCTACCACA R TCAGGGAGAAGATAAATCCGT
TLR6 F CTGCACACATCATGAAGC R TATGTTCAATGATCAGCACAGA
TLR7 F ACAACGATATCTCCACCT R CATCTCGCCATAAAACGTCT
TLR8 F CTTTATTGGGGAAAAGCA R CAAATATTTAACATGAGGCACA
TLR9 F GGAAACCAACTGAAGAGCCTGA R TTGAGCTGTCGCAACTCCC
The gene of GAPDH was used as an internal control.