Expression Of Hsp47 In Posterior Capsular Opacification

Objective: To study the expression of Hsp47 in Posterior Capsular Opacification in rabbit,and explore the role of Hsp47 in the PCO,so as to provide a new strategy and theoretical basis for regulating Hsp47 to prevent the PCO.Methods: 36 of 6-weeks health rabbit(sex randomly)into 2 groups(6 in comparison group and 30 in PCO group)randomly.Monocular PCO model was induced by Phacoemulsification the right eyes of rabbit,then executed 6 rabbits postoperation immediately,12 hours,5days,1month,3month of the operation;comparison group were executed with no operation;extirpate the eyeballs and place it on ice,remove the cornea iris and cut off the zonule ciliaire,remove the capsula and clipping a strip(2×6mm)in 4%polyformaldehyde for HE and Masson and immunohistohemical technique.the others capsulars keep in-80?.qPCR and WB was used to were employed to detect the dynamic expression of type I collagen,Hsp47,a-SMA,MMP-2,TIMP-2 protein and mRNA in the capusular of rabbit with PCO.Results:(1)HE and Masson staining: In comparison group the epithelial cells were normal in morphology and structure;after operation immediately the epithelium cells showed edema,light and loose cytoplasm,and the gaps between cells increased;the epithelium cells hyperplasia,normal epithelium and abnormal epithelium were observed in the lens at5weeks;the epithelium cells transformation,scattered small fiber tissues between cells at1month;The epithelial cells atrophic and shed to different degrees,The fiber thickness was uneven and the arrangement was irregular at 3month.(2)Immunohistohemical measurement:The Hsp47 and the type I collagen were positively expressed in PCO.In normal control group,and the expression of type I collagen and Hsp47 were weakly positive in capsular in this Comparison group..(3)QRT-PCR:The expression of Hsp47,type I collagen,a-SMA,MMP-2,TIMP-2 in the Posterior capsular opacification of rabbit were determined at mRNA level.The mRNA expression of Hsp47:Comparison group,0hr,12 hr,5days,1month and 3month they were(0.31±0.18),(0.67±0.19),(1.58±0.21),(2.07±0.29),(2.49±1.29),(6.21±2.07)respectively,there were statistical different from the contorl group(all P<0.05);The mRNAexpression of type I collagen:Comparison group,0hr,12 hr,5days,1month and3 month,Theywere(1.16±0.21),(0.79±0.37),(1.10±0.38),(1.27±0.78),(3.93±2.68),(12.2±3.91)respectively,The mRNA expression of type I collagen at 3month was statistically different from the contorl group(P<0.05).The mRNA expression of a-SMA:comparison group,0 hr,12 hr,5 days,1 month and 3 month they were(0.29±0.02),(0.36±0.05),(0.49±0.06),(1.67±0.08),(1.15±0.22),(1.09±0.03)respectively,there were statistical different from the contorl group(all P<0.05);The mRNA expression of MMP-2:comparison group,0hr,12 hr,5days,1month and 3month they were(0.25±0.12),(0.86±0.36),(2.23±0.46),(0.64±0.18),(0.48±0.07)respectively,there were statistical different from the contorl group(all P<0.05);The mRNA expression of TIMP-2:comparison group,0hr,12 hr,5days,1month and 3month they were(0.64±0.12),(0.67±0.34),(0.45±0.08),(0.82±0.17),(0.58±0.74),(7.92±0.67)respectively,there were statistical differenct from the contorl group(all P<0.05).(4)Western Blot:The expression levels of Hsp47:Comparison group,0 hr,12 hr,5days,1month,and3monththeywere(774.46±51.40),(598.67±34.11),(393.75±36.49),(1675.22±66.97),(1492±69.81),(1113.01±45.12)respectively,there were statistical different from the contorl group(all P<0.05);the expression levels of type?collagen:Comparison group,0hr,12 hr,5days,1month and 3month they were(882.57±63.62),(574.27±27.49),(311.11±63.6),(2521.91±25.67),(1719.3±36.59),(1155.53±7.46)respectively,there were statistical different from the contorl group(all P<0.05),at 5 days the expression of Hsp47 and type I collagen was significantly higher than the others group.The expression levels of a-SMA:Comparison group,0hr,12 hr,5days,1month and 3month they were(671.72±9.35),(2098.01±62.3),(1729.71±52.59),(1330.24±33.89)respectively,there were statistical different from the contorl group(all P<0.05).The expression of a-SMA protein increased immediately after surgery,and reached the highest at 5 days,and then gradually decreased.The expression levels of MMP-2:Comparison group,0hr,12 hr,5days,1month and 3month they were(556.57±15.12),(943.54±76.83),(1148.81±52.82),(2058.69±35.68),(1708.43±28.33),(1277.84±65.2)respectively,there were statistical diff-erent from the contorl group(all P<0.05);The expression levels of TIMP-2:Comparison group,0hr,12 hr,5days,1month and 3 month they were(1367.57±12.68),(843.60±30.28),(569.84±41.52),(1614.24±4.74),(746.01±44.04)respectively,there were statistical diff-erent from the contorl group(all P<0.05).The postive correlation were found in the expr-ession of HSP47 in PCO with that of type I collagen in both protein and mRNA levels-(mRNA(P<0.05,r=0.960)),protein(P<0.05,r=0.933))Conclusion:(1)Hsp47 was expressed in normal and PCO in rabbit eyes,but the expression was significantly increased in PCO.The postive correlation were found in the expression of HSP47 in PCO with that of type I collagen in both protein and mRNA levels.With time,A consistent expression trend is found between Hsp47 and type I collagen.(2)The early postoperative period(about 5 days)is the critical period for the occurrence of PCO in rabbit.This conclusion provides a theoretical basis for the optimal timing of clinical intervention the PCO.(3)MMP-2 act in the entire process of the PCO.the result that the sustained high expression of MMP-2 is the damage repair stimulation combined extracellular matrix increasy in continue,especially the type I collagen.