| Purposes of the experiment:In the experiment, by observing the relationship between skeletal musclemitochondrial energy metabolism and free radical metabolism when aging rats inaerobic exercise, and by analyzing models of aerobic exercise models of aging rats, ittries to find the senescence mechanism and seeks to explain the role of aerobicexercise on aging from the perspective of mitochondrial energy metabolism.Threerate-limiting enzymes(citroylsynthetase, isocitrate dehydrogenase, oxoglutaratedehydrogenase complex)in Tricarboxylic acid cycle, changes of activity and freeradical metabolism of Respiratory chain enzyme complex（I、II、III、IV）were analyzedin the models.Methods of the experiment:Twenty2and12-month-old healthy female Wistar rats were selected each thatbought at the Experimental Animal Center Hospital of Gansu（SCXK(Gan)2011-0001）.They should be acquired one week before the formalexperiment, and are raised by following cases: in cages in labs, keep free acess tofood and water and rodent diet by National standard, animal room temperature wascontrolled at about23℃, relative humidity was (40-60)%, natural lighting, sterilizatethe rearing room, appliances and other with UV light weekly.Firstly, put the animals in the treadmill for adaptive training one week(0°、8.2m/min、5min/day), then use of the slope50,15.2m/min in formal training,and the1st day sports15min, the2ed day30min, and to start the movement45min everydayfrom the3ed day, lasting for6days a week for12weeks.Secondly,drew the materials24hours after the12th traing week,and separately tested the activity ofcitroylsynthetase, isocitrate dehydrogenase, oxoglutarate dehydrogenase complex ofα, NADH-CoQ reductase(CI), succinatedehydrogenas（eCII）, cytochrome C reductase（CIII）, cytochrome oxidase（CIV）, Mn+-SOD、GSH-PX、CAT and the content ofMDA.Ultraviolet spectrophotometry was used in analyzing the activity ofMitochondrial respiratory chain enzyme complexesⅠ,Ⅱ,Ⅲ,Ⅳ.The kits to test the activity of citroylsynthetase, isocitrate dehydrogenase, oxoglutarate dehydrogenasecomplex of α, Mn+-SOD、GSH-PX、CAT and the content of MDA were purchasedfrom Shanghai Yuanmu Bio Co., Ltd.Results of the experiment:(1)Citroylsynthetase, isocitrate dehydrogenase, oxoglutarate dehydrogenasecomplex of α has varing degrees changes in different groups after12-week moderatetraining.Among them, the activity concentration of skeletal muscle mitochondrialcitrate synthase of group mt15is significantly higher than group mc5and mc15thatincreased by9.3%and14.2%respectively.The diffence issignificant(P<0.01).However, compared with mc5,the activity concentration of mc15has decreased by6.8%and the difference is also significant(P<0.01).The activity concentration of skeletal muscle mitochondrial isocitratedehydrogenase of group mt15is significantly higher than group mc5and mc15thatincreased by14.1%and15.3%respectively.The diffence is significant（P<0.01）.However, compared with mc5,the activity concentration of mc15hasdecreased,but there is no significant difference.Meanwhile, compared with mc5,theactivity concentration of mt5has increased by10.9%and the difference is alsosignificant(P<0.01).In contrast with group mc5and mc15, the activity concentration of skeletalmuscle mitochondrial oxoglutarate dehydrogenase complex of α of group mt15has asignificantly difference(P<0.01).And it has the same diffence by contrast mt5withmc5and mc15with mc5.(2) The results show that the impact degree of moderate exercise of aging ratsplays on skeletal muscle mitochondrial respiratory chain enzyme complexesfromⅠtoⅣ is different. Compared with group mc15and mc5, the activityconcentration of skeletal muscle mitochondrial enzyme complexes Ⅰof group mt15has increased by18.9%and12.5%which is significant(P<0.01).At the same time,group mc15is lower than mc5(P<0.05).So,there is no significant difference of theactivity concentration by compared mt5with mc5and mc15. Meanwhile,compared with group mc15and mc5, the activity concentration ofskeletal muscle mitochondrial enzyme complexesⅡof group mt15has increased by25%and14.4%which is significant(P<0.01) which has no different from mt5. Theactivity concentration of mt5increases by9%compared with mc5while thedifference is significant (P<0.05).However, the difference is not significant that groupmc15has decreased by8.5%compared with mc5.Compared with group mc15and mc5, the activity concentration of skeletalmuscle mitochondrial enzyme complexesⅢof group mt15has increased by24.1%and14%which has a sinificant difference that P<0.01.However, the difference issignificant that group mc15has decreased by8%compared with mc5and P<0.05.Theactivity concentration of mt5increases by5.2%compared with mc5while thedifference is similar.Compared with group mc15and mc5, the activity concentration of skeletalmuscle mitochondrial enzyme complexesⅣof group mt15has increased by22.7%and14.1%which has a sinificant difference that P<0.01.However,it is similar to mt5.The activity concentration of mt5increases by6%compared with mc5while thedifference is similar. Meanwhile, the difference is significant that group mc15hasdecreased by7.5%compared with mc5and P<0.05.(3)The test results also prove that the impact degree of moderate exercise ofaging rats plays on skeletal muscle radicals’related indicators is varing.The difference is significant (P<0.05) that mitochondria SOD arctivity of groupmc15increases by26%compared with mc5.The difference is the same by comparingmt15with mc15which increases by22.9%. And the index increases by54.9%bycompare mt15with mc5and the difference is significant (P<0.01).As to GSH-PX activity increases by32.6%by comparing group mc15with mc5and the difference is significant (P<0.01). Compared with mc5,the index of mc15hasa small increase and the difference is similar. It is also similar that there is a smalldecrease by comparing group mc15with mc5.Skeletal muscle mitochondria CAT activity of mc15decreases by15.2%compared with mc5and the difference is significant (P<0.05).At the same time,compared mt5with mc5,the index increases by33.8%and the difference is alsosignificant (P<0.05). By the same token,the index increases by35.5%by comparingmt15with mc15which has significant difference.however,the difference is similarcompared with mc5and the index is23%.In terms of MDA content of skeletal muscle mitochondria, the index decreasesby10.5%by comparing mt15with mc15which has significantdifference.Meanwhile,the index increases by11.7%compared with mc5.The contentof mc15increases by23.5%by compared with mc5and the difference is significant(P<0.01).There is no significant difference between mt5and mc5.Conclusions of the experiment:(1)With the aging phenomenon, body mitochondrial dysfunction, free radicaldamage and lipid peroxides increase may be the important factors of aging.(2)Exercises, especially moderate exercise, can improve the bodymitochondrial’s function and antioxidant capacity, reduce free radical’s damage andlipid peroxidation and so as to achieve the purpose of anti-aging. |
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