Effects And The Mechanism Of Reduced Glutathione On Arterial Calcification In Rats

Objective：1. Study on the establishment of rats models of the arterialcalcification and intervention, and to observe their difference on pathologicalsectionsshow.2. To oberserve the rats models of arterial calcification andintervention by the immunohistochemical method. To find the expression oftumor necrosis factor α (TNF-α) and osteoprotegerin (OPG) in the localizationand qualitative, and study the function and the significance among theglutathione (GSH), TNF-α and OPG in the process of the development of thearterial calcification.3. By using real time fluorescent quantitative PCRmethod, to find the difference of the mRNA expression of TNF-α and OPG indifferent models, and to evaluate the effect of reduced glutathione on the roleand significance of TNF-α and OPG in the process of arterial calcification.Methods:30male SD rats were randomly divided into3groups: normalcontrol group, the calcification group, the treatment group, and10rats in eachgroup. According to Price et al reported in the literature, to establish rat modelsof arterial calcification induced by Vitamin D3. 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-80℃refrigerator, for the extraction of RNA. Hematoxylin andeosin (HE) staining was used to observe the change of arterial tissue of rats atdifferent groups, Von Kossa staining (calciumstaining) can be used to see thecalcium salt deposit of arterial middle film; immunohistochemical method can find the TNF-α and OPG localization and quality in different arterial method;real time fluorescent quantitative PCR (Real Time PCR) method can be used toobserve the different group’s rat arterial TNF-alpha and OPG mRNAexpressionlevel.Results:1. See 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. The deposition of calcium salt intreatment group was obviously less than the calcification group;2. In the model of arterial calcification successfully established in rats, theimmunohistochemistry and real-time PCR results show that the protein contentand mRNA expression of TNF-αin treatment group was significantly lowerthan the calcification group(P<0.05). The immunohistochemistry and real-timePCR results of TNF-αin the control group was the lowest in the three groups(P<0.05);3. In the model of arterial calcification successfully established in rats, theimmunohistochemistry and real-time PCR results show that the protein contentand mRNA expression of TNF-αin treatment group was evidently higher thanthe calcification group(P<0.05) The immunohistochemistry and real-time PCRresults of TNF-αin the control group was the lowest in the three groups(P<0.05).Conclusions: Glutathione can inhibit the expression of TNF-α, and protectrat arterial tissue in the rat arterialcalcification, while the can produce a certaineffect on the expression of OPG by reduce the consumption of OPG.Have inhibited rat arterial calcification role as with OPG...