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Effects Of Eccentric Exhaustion Exercise And Blunt Contusion On Autophagy-related Factors Of Rat Skeletal Muscle Cells

Posted on:2020-01-12Degree:MasterType:Thesis
Country:ChinaCandidate:M J JiFull Text:PDF
GTID:2437330575993651Subject:Human Movement Science
Abstract/Summary:PDF Full Text Request
Objective:To observe the changes of autophagy-related factors Beclinl and LC3 in skeletal muscle cells of rats with centrifugal exhaustion exercise and blunt contusion,and the changes of mitochondrial autophagy-related factor PINK1 at different time points immediately after injury,24h and 48h,and to explore different injuries.The trend of related autophagy factors and the level of autophagy changes during the mode.Methods:Male 8-week-old Sprague-Dawley rats were divided into 7 experimental groups,which were quiet control group(C group),centrifugal exhaustion exercise immediate group(E0 group),24h after centrifugal exhaustion exercise group(E24 group),48h after centrifugal exhaustion exercise group(E48 group),blunt trauma immediate group(DO group),24h after blunt contusion group(D24 group),48h after blunt contusion group(D48 group),each group have 6 rats.Quiet control group:rats were narcotized by intraperitoneal injection and killed.The quadrilateral quadriceps of the hind limbs were taken and divided into two parts,one for real-time fluorescent quantitative PCR and one for Western blotting.In the blunt contusion groups and the exercise groups,SD rats were sampled by the similar steps at immediately or 24 and 48 hours after blunt contusion and exhaustive exercise.Results:(1)The results of RT-PCR of Beclinl:Compared with group C,the content of Beclinl mRNA in EO group was significantly decreased(P<0.05),and the content of Beclin1 mRNA in E24 and E48 group was not significantly changed.On the other hand,compared with group C,the content of Beclinl mRNA in group DO was higher than that in group C.There was a significant increase(P<0.01).The content of Beclinl mRNA in D24 group and D48 group was significantly decreased(P<0.05).(2)RT-PCR results of PINK1:Compared with group C,the content of PINK1 mRNA in EO group was significantly decreased(P<0.05),the change in E24 group was not significant,and the content of PINK1 mRNA in E48 group was significantly increased(P<0.05).On the other side,compared with group C,the mitochondrial PINK mRNA1 content of skeletal muscle in DO group was significantly increased(P<0.01).The skeletal muscle mitochondrial PINK1 mRNA content in D24 group was significantly increased(P<0.05).There was no significant difference in skeletal muscle mitochondrial PINK1 mRNA content in D48 group.(3)RT-PCR results of LC3:Compared with group C,and the content of LC3 mRNA in skeletal muscle cells of E24 group was not significantly changed,but the content of LC3 mRNA in skeletal muscle cells of E48 group was significantly increased(P<0.01).Compared with group C,the content of LC3 mRNA in skeletal muscle cells of group DO was significantly increased(P<0.01).There was no significant change in D24 group,and the content of LC3 mRNA in skeletal muscle cells of group D48 was significantly decreased(P<0.01).On the other hand,the content of LC3 mRNA in skeletal muscle cells of group DO was significantly higher than that of group EO(P<0.01).There was no significant difference between E24 group and D24 group,and the one in D48 group was significantly lower than that of E48 group(P<0.05)The results of Western blot of LC3 showed that compared with group C,the contents of LC3 protein of skeletal muscle in all other groups were significantly increased(P<0.05)(P<0.01).The one in E24 group was not significantly different from that in EO group,and the level of autophagy reached the highest valuein E48 group.It showed that the level of autophagy in the all stages of exhaustive exercise model gradually increased.On the other side,compared with DO group,the content of LC3 protein in skeletal muscle of D24 group was significantly increased(P<0.01),while the D48 group showed a decreasing trend compared with D24 group.In the blunt trauma model,the level of autophagy gradually increased quickly and reached the peak at 24h,then the value gradually decreases after 24hConclusions:(1)Compared with group C,the content of Beclinl mRNA,PINK1 mRNA and LC3 mRNA in the immediate group(E0)of exhaustive exercise was significantly decreased,suggesting that the exhaustive exercise recovered early and the level of autophagy was not.High;however,the levels of PINK1 mRNA,LC3 mRNA and LC3 protein in skeletal muscle cells of the 48-hour group(E48)were significantly increased,suggesting that autophagy increased in the recovery phase.(2)On the other hand,compared with group C,the content of Beclinl mRNA,PINK1 mRNA and LC3 mRNA in the immediate blunt contusion group(DO)were significantly increased,indicating immediate and early recovery after blunt contusion.The level of phagocytosis is obviously increased,and then there is a downward trend.This may be due to the difference between blunt trauma and exhaustive exercise model.The blunt contusion is more serious than exhaustive exercise injury,so its autophagy starts faster.There are also differences in the expression of autophagy-related factors at the changing phases of blunt contusion compared with exhaustive exercise injury.
Keywords/Search Tags:centrifugal exhaustion exercise, blunt trauma, skeletal muscle injury repair, Beclinl, PINK1, LC3
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