The Expression And Effects Of CTGF,TGF And SMAD Signaling Patnway During Corneal Wound Healing

Objective. To study the expression level and location of CTGF , TGF , FN protern and CTGF, type one collagen mRNA in rabbit corneal during its wound healing.To assess the interaction between CTGF and TGF and whither the Smad signaling pathway involed in this interaction.Methods 35 Albino rabbtis were chosen and divided into four groups randomly: The control group 4 eyes, Corneal ablation were performed under topical anesthesia on all tested rabbits,among them ,16 eyes were injected 15.5μgTGF antibody subconjunctivally,the other 16 eyes with 20μg Smad₄ antibody (BA1397, Boster.Co) after the ablation. CTGF, TGF and FN protern in rabbit corneas in 2h , 6h, 1d, 3d, 7d, 14d, up to 21 day and CTGF and type one collagen mRNA in rabbit corneas 6h, 3d, 7d, 21d were measured after ablation of the cornea. CTGF,TGF,and FN proterns were assessed with immunohistochemistry , CTGF and type one collagen mRAN is detected by in-situ -hybridization. Data are expressed as the mean ± SD , Statistical significance was determined with an unpaired t-test, P<0.05 was considered to be significant.Results.Corneal reepitheliarition occurred in 3.64 ± 0.43 days in the eyes only with corneal ablation ,but the TGF antibody group needed 4.89 ± 0.32 days, Smad4 antibody group needed 4.87±0.24days .The CTGF protern or mRNA did not exit in normal rabbit corneas ,but the expression of CTGF, TGF , FNproten and CTGF , type one collagen mRNA were upregulated after cornea ablation, and go up to the highest level in 3 days , 21 days after ,the expression back to the basal level.We also found that TGF antibody can reduce the expression of CTGF in cornea epithelia cells and cornea fibroblasts .but Smad4 antibody seems having no effect on this upregulatioon . Statistical analysis: The expression of TGF,the difference between the control group and the tested group is significant,the TGF antibody group or Smad4 antibody group compared with the simply wounded group ,the difference is also significant.The expression of CTGF : the difference between the control group and the tested group is significant,the difference is also significant when compared TGF antibody group with siply wounded group,but Smad4 antibody group and the simply wouded group had not significant difference.Conclusions. Injury can upregulate the the expression of CTGF, TGF , FN proterns and CTGF , type one collagen mRNA ,TGF > CTGF all affect the wound healing,TGF also can upregulate the expression of CTGF.These data support the hypothesis that TGF, CTGF promot cornea scar formation and imply that regulating CTGF > TGF synthesis may be an potent therapy target for reducing corneal scarring . Smad signaling pathway seems not involing in TGF upregulating the expression of CTGF.