The Pathological Role Of TGF-β And MMPs In The Development Of Arterial Calcification In Rats

Objective:1. To establishment of rats models of the arterialcalcification and intervention, and to observe their difference on pathological’sections show. 2.Using real time fluorescent quantitative PCRmethod, to find the difference of the mRNA expression of TGF-β and MMPs in two models, and to evaluate the effect of reduced glutathione on the role and significance of TGF-β and MMPs in the process of arterial calcification.3. Using Gene Expression Hybridization Kit, to fine the genetic variations in two group of rats calcification, and to evaluate the effect of reduced glutathione on the role and significance of genetic variations in the process of arterial calcification.Methods:60 male SD rats were randomly divided into control group and experimental group, with 30 rats in each group. By using the method of this topic researchers, Experimental group were injected Vitamin D3 insubcutaneouslyfor3consecutivedays,inordertoinducethearterialcalcificationinra tmodel. The rat aorta was divided intotwo parts, one part is made of wax, paraffin embedded blocks,used tomake paraffin section, the other part would be appropriately processed andstoraged at Liquid nitrogen, for the extraction of RNA and Gene Expression Hybridization Kit.Hematoxylin andeosin (HE) staining was used to observe the change of arterial tissue of rats in two groups, Von Kossa staining (calciumstaining) can be used to see thecalcium salt deposit of arterial lining. Real time fluorescent quantitative PCR (Real Time PCR) method can be used toobserve the different group’s rat arterial TGF-β and MMPs mRNAexpression level. Gene Expression Hybridization Kit method can be used toobserve the different group’s rat arterial genetic variationsmRNAexpression level.Conclusion:1. The Von Kossa staining results:The black dye deposition ofcalcium salts in calcification group is large and continuous. The organization ofcontrol group is integral, no black deep dyeing.2. In the model of arterial calcification successfully established in rats, the real-time PCR and Gene Expression Hybridization Kit results show that the expression of TGF-β mRNA in the control groupwas significantly lowerthan the calcification group(P<0.05).3. In the model of arterial calcification successfully established in rats, the real-time PCR and Gene Expression Hybridization Kit results show that the expression of MMPs mRNA in the control groupwas significantly lowerthan the calcification group(P<0.05).Conclusion:The arterial calcification of rats,the higher expression of TGF-β and MMPs may contribute to the formation of arterial calcification.