The Changes Of Len Structure And Transforming Growth Factor Beta1mRNA Expression In Oxidative-induced Cataract In Dogs

Objective:To determine the relationships between lens structure and expression of transforming growth factor beta1(TGF-β1)mRNA in oxidative damage cataract. Method:(1)Chosen 36 chongqing local hybrid,5～6kg weight,dogs were randomly divided into experimental group and blank control group. Among these dogs,30 dogs were divided into experimental group and 6 dogs were blank control group. Used Fenton solution injection to lens capsule set up oxidative damage cataract model. Slit lamp was used to observed dogs cataract,according to the results of slit lamp and reference to the LOCSⅢ and Kador through modified,we classificated the lens opacities Ⅰ～Ⅳ phase.(2)Selected normal control group dogs lens to make HE pathological section.(3)The lens of blank control group used real-time quantitative PCR to have TGF-β1 and GAPDH gen’s standard curve, melting curve and amplification curve. Chosed 15 dogs of experimental groups, selected Ⅰ-Ⅳ phase of cataract’s lens for real-time quantitative PCR,finally use △△Ct to detectd the expression of TGF-β1. Result:The result indicated that:(1) Experimental group during 3～6 days,the lens cortex appeared small vesicles,that was Ⅰ phase; During 9～11 days the lens vacular cell extened from equatoria to anterio cortex regions,arround or irregular loop opacity,but not influnce to observe the fundus oculi,that was Ⅱ phase;During 14-18 days An irrgular striation pointing radially from the anterior pole,which the area increased in thelens,and gradully formed a diffused gray opacity,in the opacity area observed fundus details dimnss,that’s Ⅲ phase;During 22～23 days the opacity of lens cortex were fusion,cresent shaped shadow or water-crack invisible, that’s Ⅳphase; During 25～30 days the lens cortex and nucleus totally opactiy, that’s Ⅴ phase. The lens of blank control group were transparent. (2) We can see the normal lens had a single cube lens epithelial cell under capsule membrane,the LECs size was uniform,morphological conformity,cell staining uniform,round cell connection closed. With the development of cataract, the LECs in anterior capsule and equator became abnormal from phase Ⅰ, the connection between LECs and anterior capsule became loose in phase Ⅲ and Ⅴ, the connection between posterior capsular and cortex also became loose from phase Ⅲ to Ⅴ,both anterior and posterior capsule breakup in phase Ⅴ, the cortex lens fiber around equator became loose connection and appeared fracture collapsed and toward change phenomenon in phase Ⅴ. (3) TGF-β1mRNA expression was increased in five phase of canine cataract.In phase Ⅰ TGF-β1mRNA expression is 1.29 times compared with blank control group,in phase Ⅱ TGF-β1mRNA expression is 2.72times compared with blank control group,in phaseⅢTGF-β1mRNA expression is 2.82times compared with blank control group which is highest expression group,in phase ⅤTGF-β1mRNA expression is 1.75times compared with blank control group,in phase ⅤTGF-β1mRNA expression is 1.25times compared with blank control group.TGF-β1mRNA expression was significantly increased(p<0.01) in phase Ⅱ, Ⅲ and Ⅳ compared with the blank group, phase Ⅱ and Ⅲ had significant (p<0.05) increase compared with cataract stage, the highest in phaseⅢ then phase Ⅱ. Conclusion:(1)Used Fenton solution injection to lens capsule set up cataract,easy to operate,economical and practical,take effect quickly,can be successful to established idea model of oxidative damage cataract. (2)The result showed that high expression of TGF-β1mRNA was related to LECs abnormal changed and integrity of capsule damaged.