The expression levels of cumulus expansion-related genes (HAS2, TNFAIP6, PTX3, and PTGS2) were increased in the 25 and 50 [micro]M EGCG groups but decreased in the 200 [micro]M EGCG group and the expression levels in the 50 [micro]M EGCG group were significantly higher compared to the corresponding levels in the control group (p<0.05; Figure 4).
(A) hyaluronan synthase 2 (HAS2); (B) tumor necrosis factor alpha induced protein 6 (TNFAIP6); (C) pentraxin 3 (PTX3); (D) prostaglandin 2 (PTGS2).
Primer sequences for cumulus cell expansion -related and oxidative stress-related genes Gene Primer sequences Product size (bp) GAPDH 5'-GGGTCATCATCTCTGCACCT 176 5'-GGTCATAAGTCCCTCCACGA HAS2 5'-GGATCTCCTTCCTCAGCAGTGT 106 5'-ATTCCCAGAGGTCCGCTAATG TNFAIP6 5'-TGAAAGATGGGATGCATATTGC 101 5'-CATTTGGGAAGCCTGGAGATT PTX3 5'-CATGTATGTGAATTTGGACAACGA 101 5'-GCTTGTCCCACTCGGAGTTC PTGS2 5'-CTTAAACAAGAGCATCCAGAATGG 106 5'-GCTGTACGTAGTCTTCAATCACAATCT NRF2 5'-GATGGACTTGGAGCTGCCG 140 5'-GCTCATGCTCCTTCTGTCGT SOD1 5'-CCACGTCCATCAGTTTGGAGA 92 5'-CTTTTGGCCCACCGTGTTTT CAT 5'-CTGGGACCCAACTATCTCCA 148 5'-GATGCTCGGGAGCACTAAAG GPX4 5'-GTGCTCGCTCCATGCACGA 223 5'-CCTGGCTCCTGCCTCCCA Gene Primer sequences Accession No.
Caption: FIGURE 1: The change of the coefficient of variation CV for 22 proteins: APOL6, CASP1, CNP, CXCL10, DHX58, DRAP1, DYNLT1, FAM46A, GEMIN4, GORASP1, IFITM1, IRF7, PLSCR1, RTP4, SAMD9, SAMHD1, SIGLEC1, TAF1C, TLR7, TNFAIP6, TREX1, and ZBP1, The %-axis denotes the time (unit: h), It indicates that as the critical transition occurs, that is, the time evolves towards t = 45 h (see the vertical black line), the coefficient of variation CV for 22 proteins above significantly increases in the symptomatic group and has no obvious change in the asymptomatic group, Therefore, these 22 proteins can be viewed as DNBs for early-warning signals,
Caption: FIGURE 2: The change of the indicator TPD for 22 proteins: APOL6, CASP1, CNP, CXCL10, DHX58, DRAP1, DYNLT1, FAM46A, GEMIN4, GORASP1, IFITM1, IRF7, PLSCR1, RTP4, SAMD9, SAMHD1, SIGLEC1, TAF1C, TLR7, TNFAIP6, TREX1, and ZBP1.
Real-time PCR analyses were conducted as described previously (Su et al., 2008) to quantify the mRNA levels of Has2, Ptx3, Ptgs2, Tnfaip6, and GDF9 and the housekeeping gene Rpl19 (internal control).
Effect of TSA on ECM (Has2, Ptgs2, Ptx3, and Tnfaip6) and GDF9 gene expression
The mRNA levels of the cumulus expansion-related genes Has2, Ptgs2, Ptx3, and Tnfaip6 and the expression level of GDF9 were detected.
Activation of several key ECM genes in cumulus cells plays crucial roles regulating gonadotropin-dependent cumulus expansion including Has2, Ptgs2 (also known as Cox2), Ptx3, and Tnfaip6 (Fulop et al., 2003; Lim et al., 1997; Su et al., 2003).