The Effect Of CTGF On Canine’s Ocular Filtering Bleb Scarring After Trabeculectomy And The Initial Exploration Of Effective Dosage Of MMC In Operation

Objective:(1). The amount of CTGF expressed in filter bleb organizations was detectived during filter bleb scarring process after canine ocular trabeculectomy and then researching the effection of CTFG for the filter bleb scarring after canine ocular trabeculectomy by injection of exogenous CTGF to study the role of CTGF in canine glaucoma surgery.(2). Accroding to researching the effective dosage of mitomycin(MMC) used during trabeculectomy of canines of different ages, a reference of the effective dosage of MMC was provided for clinical canine ocular trabeculectomy.Methods:(1). 12 same constitution local hybrid dogs were selected to perform the ocular trabeculectomy at their left eyes as the experimental group, and the right eyes were maintained as a control group. Two eyes of any two dogs each day were removed on the 3rd, 5th, 7th, 14 th, 21 th, 28 th day respectively. The test group clipped the bleb regional organization, the control group chipped the same regional organization. Filtering bleb organizations were clipped to made biopsy for immunohistochemistry straining, then electron microscopy was used to image and quantitative analysis.(2). 12 same constitution local hybrid dogs were selected to perform the ocular trabeculectomy at their left eyes, then these 12 dogs were randomly allocated to two groups, one was the experimental group and the other was the control group. Thereafter, the same amount of CTGF and saline(0.2ml /P) were injected under conjunctival flap, then filter blebs were observed daily to record the survival situation and the failure time. Preoperative and postoperative IOP were measured and recorded in 1, 3, 5, 7, 14, 21 days respectively.(3). Another 24 test dogs were divided to two groups: young group and elder group, 12 dogs in each group. the groups were randomly divided into A, B, C, D four groups, each group include 3 dogs whose two eyes were performed trabeculectomy surgery. After that, the concentration of MMC used in surgery in A, B, C, D groups were differ ent, they are 0.67mg/ml, 0.50mg/ml, 0.40mg/ml, 0.00 mg/ml respectively. Finally, the morphology of bleb was observed and the IOP of each dogs were measured after one month.(4).Data were analysed by Spass analysis software.Results:(1). In test group, Protein positive area ratio of CTGF Positive surface protein particles presented the change that increased then decreased as the time after surgery change. On the 3rd day it accelerated obviously,the 7th day it accelerated mostly, then it began to go down. Between the 7th postoperative day and 3rd, 5th, 7th, 14 th, 21 st, 28 th, Protein positive area ratio of CTFG protein particles’ differences were extremely significant(P <0.01). In blank control group, Protein positive area ratio of CTGF Positive surface protein particles were very small, and protein positive area ratio of CTFG protein particles’ differences were not obvious at any time point(P<0.05). At any time point after operation, Protein positive area ratio of CTFG protein particles’ differences were extremely significant between control group and test group(P <0.01). In test group, the OD value of CTFG protein particles presented the change that increased then decreased as the time after surgery change. On the 3rd day it sharply increased,the 7th day it increased mostly, then it began to go down. Between the 7th postoperative day and 3rd, 5th, 7th, 14 th, 21 st, 28 th, the OD value of CTFG protein particles’ differences were extremely significant(P <0.01). In control group, the OD value of CTFG protein particles were very low but its differences were not very obvious(P<0.05). At any time point after operation between control group and test group, the OD value of CTFG protein particles’ differences were extremely significant(P <0.01).(2). In test group which injects CTFG into conjunctival flaps of eyes, the average time about failure of filtering bleb was(13.16±0.98) d. And in control group, it was(20.05±1.05) d. The average time about failure of filtering bleb was very different between control group and test group(P <0.01). After trabeculectomy, IOP of control group and test group decreased obviously, then it rose again. In control group, IOP rose back slowly. On the 5th postoperative day it changed little, then it rose back obviously 2 weeks after operation. On the 21 st postoperative day, IOP almost rose back to the pre-operation value. By contrast, IOP of test group rose back quickly. During the first week it was the most obviously. On the 14th postoperative day, IOP rose back to the pre-operation value. Between test group and control group, IOP differences were significant at any time point after operation(P <0.01).(3). The difference between the IOP of 30 days after operation and the IOP of preoperation in each group: The difference of IOP between Group A and Croup B, Group C, Group D were significant in juvenile dogs(P <0.01). As for elder dogs, The difference of IOP between Group A, Group B and Group C, Group D were significant(P <0.01). 30 days after operation, Group A of juvenile dogs form functional filtration blebs, most of Group B, Group C and Group D form non-functional filtration blebs. Group A and Group B of old dogs form functional filtering blebs, Group C and Group D of elder dogs form non-functional filtering blebs.Conclusion:(1). To decrease the intraocular pressure, canine eye trabeculectomy always operated, because of the high rate of scar formation, effects of surgery is not ideal. In our study, CTFG was found will lead to surgery failure, so inhibiting or blocking the activity of CTGF gene may provide a new therapeutic approach to improve the success rate of glaucoma filtration surgery.(2). MMC is usually used to suppress the scar of filter bleb after trabeculectomy: the effective dosage of MMC for juvenile dogs in trabeculectomy is 0.67mg/ml and the effective dosage of MMC for elder dogs in trabeculectomy is 0.50mg/ml.