Study The Eyeball Inflammatory Damage And The Mechanism Of Retinal Cell Cytoskeleton Protein Degradation After Corneal Penetrating Injury

Objective 1. To observe inflammatory cytokines expression characteristics in the eyeball wall and the related content dynamic changes in the aqueous humor and to probing the pathogenesis of endophthalmitis after corneal penetrating injury2.To observe m-calpain,cytoskeleton protein(α-â…¡spectrin,NF200) expression characteristics in the retina, and the related content dynamic changes of degradation products of retinal cell cytoskeleton protein(α-â…¡spectrin,NF200) and m-calpain after corneal penetrating injury, and to study the mechanism of the retinal cell cytoskeleton protein(α-â…¡spectrin,NF200) degradation after corneal penetrating injury. METHODS 1. Fifty SD rats are divided into control group and experimental groups which are made corneal penetrating injury animal model in 6h,24h,48h,72h.Every group each as ten animals. Make the specimens to detect HSP90,IL-10,TNF-α,IL-1βthe histological location characteristic in the eyeball wall with immunofluorescence staining in 6h,24h,48h,72h after injury respectively; Lastly detect the related content dynamic changes of the factors in the aqueous humor by western blotting.2. Fifty SD rats are divided into control group and experimental groups which are made corneal penetrating injury animal model in 6h,24h, 48h,72h.Every group each as ten animals. Make the specimens to detect m-calpain,α-â…¡spectrin,NF200 the histological location characteristic in the retina with immunofluorescence staining in 6h,24h,48h,72h after injury respectively; Lastly detect the related content dynamic changes of the factors in the the eyeball wall tissue by western blotting.RESULTS 1. The expression of HSP90,TNF-α,IL-1βand IL-10 in normal eyeball wall and aqueous humor are at low level. HSP90 expression in aqueous humor reaches its peak within 6h after corneal penetrating injury,which was statistically significant compared with other groups (p<0.05);immunofluorescence staining results shows that it mainly expresses in the retinal cone and rod cells layer. TNF-α,IL-1βreache their peaks in 24h after the injury, which was statistically significant compared with other groups (p<0.05); The factors also can be seen in the cornea,ciliary body,retina. IL-10 reaches its peak in 72h after the injury, which was statistically significant compared with other groups (p<0.05),which can be seen mainly in the cornea and retina. 2. The expression of m-calpain in normal retina tissue is at a low level, as well as in the retinal cone and rod cells layer, but higher in the retinal ganglion and bipolar cells layer; Immunofluorescence staining results show that the expression of positive cells reaches its peak in 24h after corneal penetrating injury, then gradually decreased. Western blotting results show that the expression of m-calpain in the eyeball wall tissue gradually increased, reaches its peak in 24h after corneal penetrating injury, which was statistically significant compared with other groups (p<0.05). Immunofluorescence staining results show thatα-â…¡spectrin and NF200 distribute in the processes of visual cells of normal retina tissue, which arrange in parallel, while the processes distribute dispersed and its realm unclear in 24h after corneal penetrating injury indicated protein degradation in the processes. Western blotting results showed that the expression ofα-â…¡spectrin and NF200 degradation products which increased as the expression of m-calpain increased and reaches their peaks in 24h after corneal penetrating injury, which was statistically significant compared with other groups (p<0.05).Conclusions 1. The stress after corneal penetrating injury can promot the local expression of inflammatory cytokines,as a result, leading a secondary inflammation injury.We should use immunosuppressive drugs early to inhibit the local inflammatory response after corneal penetrating injury.2. Local inflammatory response leading to Ca²⁺ overload in the retinal cells can activate the calcium-activated neutral protease (m-calpian).As an injury factor, the inhibition of m-calpian activity can protect cytoskeleton protein(α-â…¡spectrin,NF200) of the retinal cell to be degraded.