| Purpose:The primary aim of this study is observing the apoptosis' s changes of rat's muscle which had syndrome of blood stasis after injury.we also made analysis on the concentration changes of factors such as caspase3?caspase8?DRP1?FADD?RIP1 and RIP.we studied the pathological process of blood stasis syndrome in microcosmic way.we found that the fact of blood stasis syndrome is apoptosis.Necrotic apoptosis and physiological apoptosis were both in the processing of blood stasis syndrome.they competed with each other,which is the result of damagers and repairers after injury.Method:1.Modeling methodThe model group of the injury blood stasis symptom is completed by the modeling method of quantitative gravity strike.2.Sectionalization80 Wistar rats were divided into eight groups using table of random numbers.group A(blank control),group B(1h after molding),group C(12h after molding),group D(24h after molding),group E(72h after molding),group F(120h after molding),group G(168h after molding),group H(336h after molding)?3.observationUsing Tunel assay and Elisa assay,detection the concentration changes of factors caspase3?caspase8?DRP1?FADD?RIP1 and RIP on group A(blank control),group B(1h after molding),group C(12h after molding),group D(24h after molding),group E(72h after molding),group F(120h after molding),group G(168h after molding),group H(336h after molding)?4.Statistical analysis of dataGraph Pad Prism 6 software was used for graphic analysis.all of the Experimental data were analyzed by using the SPSS 19.0 software(IBM,Armonk,NY,USA).Measurement data were expressed with the mean-standard deviation.Comparisons of data between 2 groups were performed by using one-way ANOVA.The LSD-t method was used for homogeneity of variance.heterogeneity of variance using the way of Tamhane'sT2.Reslut:1.Using Tunel assay on the death of skeletal muscle cell we found that Statistically significant differences were found in group D(24h after molding),and group G(168h after molding)(P<0.05).the others had no significant differences.2.Using Elisa assay on the death of skeletal muscle cell we found that:1)Caspase-3 levels between the trauma and control groups(P<0.05).Further comparisons between the different groups at each time point were performed.Caspade-3 levels in the trauma groups were higher than those in the control groups at 1 hour(P<0.05),and then reduced after the quantitative gravity strike 12 hours gradually(P<0.05).The cellular apoptosis reached to the platform period.After the injury(3rd day),cellular apoptosis appeared a sharp change.There was a significant difference between the control group and trauma group in the skeletal muscle Cspase-3 levels at 3rd days and 7th days after the injury(P<0.05).This change lasted to the 14 th day2)Caspase8 levels between the trauma and control groups(P<0.05).Further comparisons between the different groups at each time point were performed.Statistically significant differences were found between group A(blank control),compare to group B(1h after molding),group C(12h after molding),group D(24h after molding),group E(72h after molding),group F(120h after molding),group G(168h after molding),and group H(336h after molding).and also found in group D(24h after molding)compare to group E(72h after molding),group E(72h after molding)compare to group F(120h after molding),group E(72h after molding)compare to group F(120h after molding).3)DRP1 levels between the trauma and control groups(P<0.05)Statistically significant differences were found between group A(blank control),compare to group B(1h after molding),group C(12h after molding),group D(24h after molding),group E(72h after molding),group F(120h after molding),group G(168h after molding),and group H(336h after molding).and also found in group C compare to group D,group D compare to group E.4)Statistically significant differences were found in the Skeletal muscle FADD levels between the trauma and control groups(P<0.05).Further comparisons between the different groups at each time point were performed.FADD levels in the trauma groups were higher than those in the control groups at 48 th hour(P<0.05),and then reduced after the quantitative gravity strike 72 hours gradually(P<0.05).This change lasted to the 14 th day5)Statistically significant differences were found in the Skeletal muscle RIP-1,levels between the trauma and control groups(P<0.05).Further comparisons between the different groups at each time point were performed.RIP-1levels had the similar change after the injury.The proteins in the trauma groups were lower than those in the control groups in 1st hour(P<0.05),and then increased after quantitative gravity strike 12 hours gradually(P<0.05).This change lasted to the 14 th day6)Statistically significant differences were found in the Skeletal muscle RIP-3 levels between the trauma and control groups(P<0.05).Further comparisons between the different groups at each time point were performed.RIP-3 levels had the similar change after the injury.The protein in the trauma groups were lower than those in the control groups in 1st hour(P<0.05),and then increased after quantitative gravity strike 12 hours gradually(P<0.05).This change lasted to the 14 th dayConclussion:1.The fact of blood stasis syndrome after injury is celluar apoptosis but not necroptosis.2.The process of celluar apoptosis in blood stasis syndrome after injury is significant temporal disciplinary:0-24 h is the incubation period;at 24 h,it's the bursting point;24-168 h is the plateau period;at 168 h it's the inflection point;the restoration stage of apoptosis is after 168h3.Blood stasis syndrome after injury is a Necrotic process with both apoptosis and necroptosis in blood stasis syndrome after injury,the factors DRP1?RIP1?RIP3 were closely related to necroptosis4.Caspase8 had some special functions in the blood stasis syndrome after injury5.The apoptosis pathway of blood stasis syndrome after injury may not go through Caspase8,and the reduce of Caspase8 level may be the reason of necroptosis. |
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