雄激素經(jīng)IGF-I介導(dǎo)調(diào)節(jié)運動骨骼肌mTOR信號機理的探索
發(fā)布時間：2018-09-11 10:14
【摘要】：研究目的：本研究通過不同強度運動及應(yīng)用IGF-I和雄激素,觀察運動骨骼肌AR、MAPK和mTOR通路間的變化特點與規(guī)律,對雄激素經(jīng)IGF-I介導(dǎo)調(diào)節(jié)運動骨骼肌mTOR信號機理進行探討。 研究方法：實驗一雄性SD大鼠進行不同強度運動(跑臺運動：速度20min/min和26m/min、60min、10%坡度),運動后即刻、6h和12h取材白腓腸肌。實驗二雄性SD大鼠肌肉注射IGF-I,注射后20、60和120min取白腓腸肌。實驗三將雄性SD大鼠分為安靜對照組(S)、DHT組(D)、運動組(E)和DHT運動組(ED),D和ED組頸背部皮下埋植DHT緩釋劑,其余假手術(shù)。E和ED組進行以20m/min、60min、10%坡度的跑臺運動。再細分為三天組和十天組,運動后12h取白腓腸肌。BCA法測肌肉蛋白濃度,HE染色觀察骨骼肌形態(tài),實時熒光定量RT-PCR測肌肉AR和IGF-I基因表達,Western Blot法測肌肉MHC和AR、MEK、ERK、p90RSK、 mTOR、p70S6K和4EBP1的磷酸化。 研究結(jié)果：(1)急性運動時,骨骼肌AR、MAPK和mTOR通路磷酸化狀態(tài)存在運動強度依賴性,且MAPK通路活性在運動后即刻出現(xiàn)峰值,AR和mTOR通路的出現(xiàn)于6小時。(2)肌肉注射外源性IGF-I增強了肌肉AR表達,AR活性于注射后60min達峰值。骨骼肌MAPK和mTOR通路活性增強,均于注射后20min達到峰值。(3)皮下埋植DHT使肌肉橫斷面、濕重和MHC明顯增加。運動加強了雄激素的促合成效應(yīng)。(4)DHT作用三天明顯增加肌肉內(nèi)IGF-I含量,而DHT結(jié)合運動后能進一步提高肌肉內(nèi)IGF-I。(5)皮下埋植DHT使MAPK和mTOR通路的表達均顯著增強。 研究結(jié)論：(1)骨骼肌AR、MAPK通路和mTOR通路的活性存在強度依賴性。急性運動后骨骼肌MAPK通路峰值出現(xiàn)較早,AR和mTOR通路峰值較晚。(2)雄激素明顯促進了骨骼肌的肥大,還增強了肌肉IGF-I生成。運動能加強了雄激素的促肌肉合成效應(yīng)。同時,IGF-I除促進骨骼肌AR的表達及其活性外,還能迅速促進MAPK通路和mTOR通路的活性。(3)骨骼肌MAPK通路和mTOR通路均為雄激素的非基因途徑,且MAPK通路位于mTOR通路的上游。
[Abstract]:Objective: to observe the changes of AR,MAPK and mTOR pathway in skeletal muscle by using IGF-I and androgen at different intensities, and to explore the mechanism of mTOR signal regulation by IGF-I mediated by androgen. Methods: male SD rats underwent different intensity exercises (treadmill movement: speed 20min/min and 26 m / min 60 min / min 10% slope). The white gastrocnemius muscle was obtained immediately after exercise for 6 h and 12 h. In experiment 2, the white gastrocnemius muscle was taken from male SD rats by intramuscular injection of IGF-I, (20: 60 and 120min). In experiment 3, male SD rats were divided into control group, (D), group, (E) group, DHT exercise group, (ED) D group and ED group, where DHT was implanted subcutaneously into the neck and back. The rest of sham-operated .E and ED groups were treated with 10% slope treadmill exercise at 20 m / min for 60 min. After 12 hours exercise, the muscle protein concentration was measured by HE staining, and the expression of AR and IGF-I genes in muscle was measured by real-time fluorescence quantitative RT-PCR. The phosphorylation of MHC, AR,MEK,ERK,p90RSK, mTOR,p70S6K and 4EBP1 in muscle was detected by Western Blot. Results: (1) Phosphorylation of AR,MAPK and mTOR pathways in skeletal muscle was intensity-dependent during acute exercise. The activity of MAPK pathway appeared peak at 6 hours after exercise. (2) exogenous IGF-I increased the activity of AR expression in muscle and reached the peak value of 60min after injection. The MAPK and mTOR pathway activity of skeletal muscle increased, both of which reached the peak value after injection. (3) Subcutaneous implantation of DHT significantly increased muscle cross-section, wet weight and MHC. Exercise enhanced the effect of androgen synthesis. (4) DHT significantly increased the content of IGF-I in muscle for three days, while DHT combined with exercise could further increase the expression of MAPK and mTOR pathway after subcutaneous implantation of DHT (5) in muscle. Conclusion: (1) the activities of AR,MAPK pathway and mTOR pathway in skeletal muscle are intensity-dependent. The peak value of MAPK pathway in skeletal muscle after acute exercise was earlier than that in AR and mTOR pathway. (2) androgen significantly promoted skeletal muscle hypertrophy and increased muscle IGF-I production. Exercise boosts the effect of androgen on muscle synthesis. At the same time, IGF-I not only promoted the expression and activity of AR in skeletal muscle, but also increased the activity of MAPK pathway and mTOR pathway. (3) the MAPK pathway and mTOR pathway of skeletal muscle were both non-androgen pathways, and MAPK pathway was located upstream of mTOR pathway.
【學(xué)位授予單位】：北京體育大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2012
【分類號】：G804.2
本文編號：2236396
[Abstract]:Objective: to observe the changes of AR,MAPK and mTOR pathway in skeletal muscle by using IGF-I and androgen at different intensities, and to explore the mechanism of mTOR signal regulation by IGF-I mediated by androgen. Methods: male SD rats underwent different intensity exercises (treadmill movement: speed 20min/min and 26 m / min 60 min / min 10% slope). The white gastrocnemius muscle was obtained immediately after exercise for 6 h and 12 h. In experiment 2, the white gastrocnemius muscle was taken from male SD rats by intramuscular injection of IGF-I, (20: 60 and 120min). In experiment 3, male SD rats were divided into control group, (D), group, (E) group, DHT exercise group, (ED) D group and ED group, where DHT was implanted subcutaneously into the neck and back. The rest of sham-operated .E and ED groups were treated with 10% slope treadmill exercise at 20 m / min for 60 min. After 12 hours exercise, the muscle protein concentration was measured by HE staining, and the expression of AR and IGF-I genes in muscle was measured by real-time fluorescence quantitative RT-PCR. The phosphorylation of MHC, AR,MEK,ERK,p90RSK, mTOR,p70S6K and 4EBP1 in muscle was detected by Western Blot. Results: (1) Phosphorylation of AR,MAPK and mTOR pathways in skeletal muscle was intensity-dependent during acute exercise. The activity of MAPK pathway appeared peak at 6 hours after exercise. (2) exogenous IGF-I increased the activity of AR expression in muscle and reached the peak value of 60min after injection. The MAPK and mTOR pathway activity of skeletal muscle increased, both of which reached the peak value after injection. (3) Subcutaneous implantation of DHT significantly increased muscle cross-section, wet weight and MHC. Exercise enhanced the effect of androgen synthesis. (4) DHT significantly increased the content of IGF-I in muscle for three days, while DHT combined with exercise could further increase the expression of MAPK and mTOR pathway after subcutaneous implantation of DHT (5) in muscle. Conclusion: (1) the activities of AR,MAPK pathway and mTOR pathway in skeletal muscle are intensity-dependent. The peak value of MAPK pathway in skeletal muscle after acute exercise was earlier than that in AR and mTOR pathway. (2) androgen significantly promoted skeletal muscle hypertrophy and increased muscle IGF-I production. Exercise boosts the effect of androgen on muscle synthesis. At the same time, IGF-I not only promoted the expression and activity of AR in skeletal muscle, but also increased the activity of MAPK pathway and mTOR pathway. (3) the MAPK pathway and mTOR pathway of skeletal muscle were both non-androgen pathways, and MAPK pathway was located upstream of mTOR pathway.
【學(xué)位授予單位】：北京體育大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2012
【分類號】：G804.2
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