The Differentially Expressive Research Of Protein And Proteome Of Gastrocnemius After Acute Exhaustive Exercise On Rats

Object: In order to study the differential expressive characteristic of exercise rats gastrocnemius proteome, and to explore the mechanism of exercise-induced fatigue on the proteomic level. Method: Ten male SD rats were divided into two groups randomly according to their weight: the rest group (5) and the exercise group (5). The exercise group was sent to a one-time acute exhausting treadmill training. They were executed by decollation at the third hour after exhausted exercise. The rest group was executed at the same time. The entire proteins of back limb gastrocnemius muscle were extracted and divided into two experiment. Section One: The primary study of the Differential Expressive proteome of Rats' gastrocnemius after Acute Exercise. The proteins were separated by SDS-PAGE, stained, digested in gel with trypsin, and then analyzed by ESI-MS. Section Two: The Comparative Proteomic Study of Rats' Skeletal Muscle after Acute Exhaustive Exercise. The proteins were separated by 2-DE, stained by silver nitrate, digested in gel with trypsin, analyzed by MALDI-TOF-TOF-MS in turn. Results of Section One: The protein extraction was successful, and the concentration of protein was 40ug.ul^-1, which was determined by Bradford method. There are 36 bands of each lane. Some bands are significantly wider and darker, while some bands are more narrow and more lighter, compared with the rest group.The significantly changed bands were digested by enzyme in gel and analyzed by MS and revealed that the exercise group contains alpha-fetoprotein in the third band. Result of Section Two: 572 ± 28 protein spots were found in the trained group and 667 ±35 in the rest group. In the 282 differentially expressed spots, after training, 88 spots disappeared, 32 spots appeared, 12 spots increased to twice the volume, and 10 spots decrease to half the volume. Seven differential expressive proteins were identified by MALDI-TOF-TOFMS. They are glyceraldehyde 3-phosphate-dehydrogenase(disappeared),H+-transporting two-sector ATPase(disappeared), glyoxylase 1 (appeared), myosin light polypeptide 3(down-expressed), phosphoglucomutase 1 (down-expressed), acyl-Coenzyme A dehydrogenase(disappeared)and two unknown proteins (up-expressed). Conclusion: The disappeared and down-regulated proteins in exercise group are related to energy metabolization and disturbance of muscle contract regulating and exercise capability decreasing and exercise-induced fatigue. The additional and up -regulated proteins may be related to tissue damage and immunity inhibition. The high sensitive and differential result in exercise stress condition reveals that, using proteomic strategy and technology to study reflection and adoption and exercise-induced task, have important meaning and wide prospect.