The primer sequences used were as follows: Human Actin: 5-AGA GCT ACG AGC TGC CTG AC-3' and 5'-AGC ACT GTG TTG GCG TAC AG-3'; Human TLR2: 5'-GCC AAA GTC TTG ATT GAT TGG-3' and 5'-TTG AAG TTC TCC AGC TCC TG-3'; Human Collagen III: 5'-TGG TGT TGG AGC CGC TGC CA-3' and 5'-CTC AGC ACT AGA ATC TGT CC-3'; Human Collagen IV: 5'-ATG TCA ATG GCA CCC ATC AC-3' and 5'-CTT CAA GGT GGA CGG CGT AG-3'; Human CatB: 5'-TGA CGT GTT GGT ACA CTC CTG-3' and 5'-TGG AGG GAG CTT TCT CTG TG-3'; Mouse Actin: 5'-CAATAG TGATGA CCT GGC CGT-3' and 5'-AGA GGG AAA TCG TGC GTG AC-3'; Mouse Collagen III: 5'-CCA GCT GGG CCT TTG ATA CCT-3' and 5'-TGC CCA CAG CCT TCT ACA CCT-3'; and Mouse Collagen IV: 5'-AGG CAG GTC AAG TTC TAG CG-3' and 5'-CAA GCA TAG TGG TCC GAG TC-3'.
The Expression of CatB as well as Collagen III and Collagen IV after P.g.
LPS (1 [micro]g/mL), the mean mRNA expression of CatB was significantly increased at 24 and 48 h (the late culture periods) but not at 3 or 12 h (the early culture periods) compared with the unchallenged cells (Figure 1(b)).
The Regulation of Collagens III and IV Expression by CatB in Fibroblasts after P.g.
These observations clearly demonstrate that CatB has a critical role in regulating the expression of collagens III and IV by fibroblasts during chronic activation of TLR2 signaling.
The Regulation of NF-[kappa]B Activation and Oxidative Damage for Decreasing Collagens III and IV by CatB after P.g.
We then further analyzed the expression of CatB, I[kappa]B[alpha], and 8-OHdG (a critical biomarker of oxidative stress).
These observations demonstrate that CatB is involved in the proteolytic degradation of the I[kappa]B[alpha] and oxidative DNA damage during chronic P.g.
Determination of CatB and Oxidative Damage in Fibroblasts of Inflamed Tissues with Chronic Periodontitis.
The major finding of the present study was clarifying the critical role of CatB in regulating collagen expression by fibroblasts via prolonging TLR2/NF-[kappa]B activation (summarized in Figure 5).
Elevated levels of CatB in fibroblasts are typically observed in many chronic inflammatory diseases, including rheumatoid arthritis as well as periodontitis [32-34].
CatB was recently found to regulate NF-[kappa]B activation .