本文選題：AR + 氟他胺　； 參考：《上海體育學(xué)院》2017年碩士論文

【摘要】：研究目的:雄激素與雄激素受體(androgen receptor,AR)結(jié)合后有促進(jìn)肌肉蛋白質(zhì)合成、增加肌肉質(zhì)量和提高肌肉力量的作用,但其作用機(jī)制仍未明確。本課題以SD雄性大鼠為研究對象,利用AR阻斷劑Flutamine(氟他胺)研究AR對安靜和運(yùn)動大鼠運(yùn)動能力的影響及其機(jī)制-是否與胰島素樣生長因子1(insulin growth factor 1,IGF-1)/磷脂酰肌醇3激酶(phosphoinositide 3-kinase,PI3K)/Akt/哺乳動物雷帕霉素靶蛋白(mammalian target of rapamycin,m TOR)通路有關(guān)?研究方法:48只SD雄性大鼠(7周齡)隨機(jī)分為6組,每組8只:安靜對照組(C)、氟他胺組(F)、抗阻訓(xùn)練組(R)、抗阻訓(xùn)練+氟他胺組(R+F)、耐力訓(xùn)練組(E)和耐力訓(xùn)練+氟他胺組(E+F)。實驗前將氟他胺21天緩釋劑(50 mg/片)包埋于F、R+F和E+F組大鼠的頸部皮下。C組和F組大鼠不進(jìn)行運(yùn)動訓(xùn)練,而E、E+F組大鼠進(jìn)行中等強(qiáng)度(速度20 m/min)的跑臺耐力訓(xùn)練,R、R+F組大鼠進(jìn)行遞增負(fù)荷的爬梯抗阻訓(xùn)練,均為每周訓(xùn)練6天,持續(xù)3周。最后一次訓(xùn)練后36 h,檢測所有大鼠的體重、力量(最大抓力和啟動時最大負(fù)重分別反映最大力量和速度力量)和耐力(一次力竭運(yùn)動時間)。在力竭運(yùn)動結(jié)束36 h后,麻醉大鼠,下腔靜脈取血。之后,處死大鼠,取腓腸肌、比目魚肌。ELISA方法檢測血清IGF-1水平;蒽酮法檢測骨骼肌的肌糖原含量,real time PCR檢測骨骼肌IGF-1R和m TOR的m RNA水平;Western blot檢測骨骼肌的AR、IGF-1、IGF-1R、m TOR、PI3K、肌球蛋白重鏈(Myosin heavy chain,MHC)、快肌球蛋白(fs-myosin)和慢肌鈣蛋白(ss-Tn T)的蛋白水平,以及PI3K和Akt的表達(dá)和活性。研究結(jié)果:1.AR介導(dǎo)安靜和抗阻訓(xùn)練大鼠體重的增加,但耐力訓(xùn)練所致大鼠體重的減輕與AR無關(guān)氟他胺不僅降低安靜對照組大鼠的體重(p0.01),還降低抗阻訓(xùn)練大鼠的體重(p0.05),但對耐力訓(xùn)練所致大鼠體重的減輕無顯著影響(p0.05)。2.AR介導(dǎo)抗阻訓(xùn)練所致大鼠最大力量和速度力量的增加,但耐力訓(xùn)練增加耐力的作用與AR無關(guān)抗阻訓(xùn)練而不是耐力訓(xùn)練顯著增加大鼠的最大力量(最大抓力/體重比值)和速度力量(啟動時最大負(fù)重)(p0.05),且該作用被氟他胺逆轉(zhuǎn)(p0.05)。而耐力訓(xùn)練增加大鼠耐力的作用(p0.05)不能被AR減弱,表明其與AR無關(guān)。3.抗阻訓(xùn)練大鼠腓腸肌和耐力訓(xùn)練大鼠比目魚肌相對重量的增加是通過AR介導(dǎo)的抗阻訓(xùn)練增加中間態(tài)肌腓腸肌的相對重量(p0.05),且該作用被氟他胺逆轉(zhuǎn)(p0.01),但對比目魚肌的相對重量無影響。而耐力訓(xùn)練增加慢肌比目魚肌的相對重量(p0.05),且該作用被氟他胺逆轉(zhuǎn)(p0.01),但對腓腸肌的相對重量無影響。4.抗阻訓(xùn)練通過AR介導(dǎo)大鼠腓腸肌快肌纖維的增多,而耐力訓(xùn)練大鼠比目魚肌慢肌纖維的增多與AR無關(guān)抗阻運(yùn)動增加腓腸肌快肌特異性蛋白fs-myosin的含量(p0.05),對慢肌特異性蛋白ss Tn T無顯著作用;抗阻運(yùn)動增加腓腸肌fs-myosin含量的作用被氟他胺逆轉(zhuǎn)(p0.01)。而耐力訓(xùn)練增加比目魚肌ss Tn T的含量(p0.05),但對fs-myosin含量無顯著作用。耐力訓(xùn)練增加比目魚肌ss Tn T含量的作用不被氟他胺減輕。5.抗阻訓(xùn)練大鼠腓腸肌和耐力訓(xùn)練大鼠比目魚肌MHC含量的增加是通過AR介導(dǎo)的抗阻訓(xùn)練增加大鼠腓腸肌的MHC蛋白水平(p0.05),耐力訓(xùn)練增加比目魚肌的MHC蛋白水平(p0.05),且運(yùn)動引起的MHC蛋白水平增加的作用都能被氟他胺所逆轉(zhuǎn)(p0.01)。6.抗阻和耐力運(yùn)動引起的大鼠腓腸肌和比目魚肌肌糖原含量的增加是通過AR介導(dǎo)的與安靜對照大鼠相比,抗阻和耐力訓(xùn)練都能顯著增加腓腸肌和比目魚肌的肌糖原含量(p0.01),而運(yùn)動增加肌糖原含量的作用被氟他胺逆轉(zhuǎn)(p0.01)。7.抗阻訓(xùn)練大鼠腓腸肌和耐力訓(xùn)練大鼠比目魚肌IGF-1R/m TOR通路信號分子的增加通過AR介導(dǎo),而抗阻和耐力訓(xùn)練不影響血清IGF-1水平抗阻訓(xùn)練顯著增加腓腸肌、耐力訓(xùn)練顯著增加比目魚肌的IGF-1R和m TOR的m RNA水平(均p0.05),且該作用都能被氟他胺逆轉(zhuǎn)(p0.01,p0.05)。對于蛋白水平,抗阻訓(xùn)練增加腓腸肌IGF-1(p0.05)、IGF-1R(p0.01)和m TOR(p0.05)的蛋白水平,該作用被氟他胺逆轉(zhuǎn)(p0.05,p0.01,p0.01);而耐力訓(xùn)練增加比目魚肌IGF-1(p0.01)和IGF-1R(p0.05)蛋白水平,該作用被氟他胺逆轉(zhuǎn);此外,氟他胺降低降低比目魚肌的m TOR(p0.05)蛋白水平。此外,盡管抗阻和耐力訓(xùn)練未影響血清IGF-1的水平,以及耐力訓(xùn)練不影響比目魚肌的m TOR的蛋白水平,但上述指標(biāo)均能被氟他胺降低。8.抗阻訓(xùn)練大鼠腓腸肌和耐力訓(xùn)練大鼠比目魚肌PI3K和Akt的表達(dá)和活性的增加是AR介導(dǎo)的抗阻訓(xùn)練增加腓腸肌、耐力訓(xùn)練增加比目魚肌的PI3K和Akt的活性(p-PI3K/PI3K、p-Akt/Akt的比值)(均p0.05),該作用被氟他胺逆轉(zhuǎn)(均p0.05)。結(jié)論1.抗阻訓(xùn)練通過AR介導(dǎo)大鼠最大力量和速度力量的增強(qiáng),而耐力訓(xùn)練提高耐力的作用可能與AR無關(guān)。2.抗阻訓(xùn)練通過AR介導(dǎo)大鼠力量增強(qiáng)的機(jī)制與其增加腓腸肌的相對重量、快肌纖維含量、內(nèi)在收縮力和肌糖原含量有關(guān)。3.耐力訓(xùn)練通過AR介導(dǎo)比目魚肌相對重量、內(nèi)在收縮力和肌糖原含量的增加,但其增加慢肌纖維的作用不是通過AR介導(dǎo)。這可能是AR阻斷劑在降低比目魚肌相對重量、收縮力和肌糖原含量的情況下沒有降低大鼠耐力的可能機(jī)制之一,但仍需進(jìn)一步證實。4.抗阻訓(xùn)練增加腓腸肌、耐力訓(xùn)練增加比目魚肌相對重量的機(jī)制與運(yùn)動激活肌肉AR,進(jìn)而激活I(lǐng)GF-1/IGF-1R-PI3K/Akt-m TOR信號通路有關(guān)。但是,訓(xùn)練通過AR的介導(dǎo)如何增加快肌纖維含量、MHC水平和肌糖原含量,仍待研究。
[Abstract]:Objective: the combination of androgen and androgen receptor (androgen receptor, AR) can promote muscle protein synthesis, increase muscle mass and improve muscle strength, but its mechanism is still not clear. This subject takes SD male rats as the research object, and uses the AR blocker Flutamine (fluretamine) to study the exercise of AR on the exercise of quiet and exercise rats. The effect of ability and its mechanism - is it related to the insulin like growth factor 1 (insulin growth factor 1, IGF-1) / phosphatidyl inositol 3 kinase (phosphoinositide 3-kinase, PI3K) /Akt/ mammal rapamycin target protein (mammalian target of rapamycin) pathway? Methods: 48 male rats (7 weeks of age) were randomly divided into 6 groups, each Group 8: quiet control group (C), flualamine group (F), resistance training group (R), resistance training + flutamide group (R+F), endurance training group (E) and endurance training + flutamide group (E+F). Before experiment, the 21 day slow-release flualamines (50 mg/ tablets) were embedded in F, R+F and E+F group rats' neck subcutaneous.C group and F group rats were not exercise. Medium strength (speed 20 m/min) endurance training, R and R+F rats were trained for incremental load of climbing ladder resistance training for 6 days a week for 3 weeks. After the last training, 36 h was used to detect the weight of all rats. Strength (maximum grasp and maximum weight reflected the maximum strength and speed strength) and endurance (one exhaustion). Exercise time). After the exhaustion movement ended 36 h, the rats were anesthetized and the inferior vena cava was taken blood. After that, the rats were killed, the gastrocnemius muscle, the soleus muscle and the.ELISA method were used to detect the serum IGF-1 level. The content of the skeletal muscle glycogen was detected by the anthrone method, and the real time PCR was used to detect the m RNA level of the skeletal muscle IGF-1R and m TOR. 1R, m TOR, PI3K, myosin heavy chain (Myosin heavy chain, MHC), the protein levels of fast myosin (fs-myosin) and slow troponin (ss-Tn T), as well as the expression and activity of PI3K and ss-Tn. The weight (P0.01) of the rats in the low quiet control group and the weight of the resistance training rats (P0.05) had no significant effect on the weight loss of rats induced by endurance training (P0.05).2.AR mediated the increase of maximum strength and speed of rats induced by resistance training, but endurance training was not related to AR, but not endurance training, not endurance. Training significantly increased the maximum strength (maximum grip / weight ratio) and speed strength (P0.05), and the effect was reversed by flutamide (P0.05). The effect of endurance training increased rat endurance (P0.05) could not be weakened by AR, indicating that it was not related to AR in the gastrocnemius and endurance training rat soleus with.3. resistance training. The increase in relative weight of the muscle increased the relative weight of the meso gastrocnemius by AR mediated resistance training (P0.05), and the effect was reversed by flutamide (P0.01), but had no effect on the relative weight of the soleus muscle. Endurance training increased the relative weight of the slow soleus muscle (P0.05), and the effect was reversed by flutamide (P0.01), but to the gastrocnemius. The relative weight of the muscle did not affect the increase of the.4. resistance training through AR, while the increase of the slow muscle fiber in the endurance training rat and the AR independent resistance exercise increased the content of the fast muscle specific protein fs-myosin of the gastrocnemius muscle (P0.05), and had no significant effect on the slow muscle specific protein SS Tn T, and the resistance exercise increased. The effect of fs-myosin content of gastrocnemius was reversed by flutamide (P0.01). Endurance training increased the content of SS Tn T in soleus muscle (P0.05), but had no significant effect on fs-myosin content. The effect of endurance training to increase the content of SS Tn T in soleus muscle was not reduced by flutamide in reducing the MHC content of the gastrocnemius and endurance training rat soleus muscle of.5. resistance training rats. The increase in AR mediated resistance training increases the MHC protein level (P0.05) of the gastrocnemius muscle in rats and the endurance training increases the MHC protein level of the soleus muscle (P0.05), and the effect of the increase of MHC protein in the exercise can be caused by the reverse (P0.01).6. resistance and endurance exercise of the gastrocnemius and soleus muscle glycogen caused by the exercise of the resistance and endurance exercise of the (P0.01).6.. The increase in content is that resistance and endurance training can significantly increase the content of muscle glycogen (P0.01) in the gastrocnemius and soleus muscle by AR mediated resistance and endurance training, while the action of increasing the content of muscle glycogen in the exercise is the IGF-1R/m TOR pathway of the gastrocnemius and the endurance training rat of P0.01.7. resistance training. The increase of signal molecules was mediated by AR, while resistance and endurance training did not affect the sera IGF-1 level resistance training significantly increasing the gastrocnemius muscle, endurance training significantly increased the m RNA level of IGF-1R and m TOR of the soleus muscle (P0.05), and the effect could be reversed by flutamide (P0.01, P0.05). For protein level, resistance training increased the IGF-1 gastrocnemius muscle (P0.05) the protein levels of IGF-1R (P0.01) and m TOR (P0.05), which were reversed by fluamamines (P0.05, P0.01, P0.01), and endurance training increased the level of IGF-1 (P0.01) and IGF-1R protein in the soleus, and the effect was reversed by flutamide; in addition, flutamide decreased the protein level of the soleus muscle. In addition, resistance and endurance, in spite of resistance and endurance. The training did not affect the level of serum IGF-1, and endurance training did not affect the protein level of M TOR in the soleus muscle, but the above indexes could be reduced by fluamamines to decrease the expression and activity of PI3K and Akt in the gastrocnemius and endurance training rats of.8. resistance training. The increase of the gastrocnemius muscle and endurance training increased by AR mediated resistance training. The activity of PI3K and Akt in soleus muscle (the ratio of p-PI3K/PI3K, p-Akt/Akt) (both P0.05), the effect was reversed by flutamide (all P0.05). Conclusion 1. resistance training is mediated by AR to enhance the strength and strength of the rats' maximum strength and speed, and the effect of endurance training to improve endurance may be associated with AR independent.2. resistance training through AR mediated rat strength enhancement machine. The relative weight of the gastrocnemius muscle, the fast muscle fiber content, the intrinsic contractile force and the muscle glycogen content related.3. endurance training through AR mediate the relative weight of the soleus, the internal contractile force and the muscle glycogen content, but the effect of the increase of the slow muscle fiber is not mediated by the AR. This may be that the AR blocker is relative to the soleus muscle. Weight, contractile force and muscle glycogen content is not one of the possible mechanisms to reduce rat endurance, but it is still necessary to further confirm that.4. resistance training increases the gastrocnemius muscle, endurance training increases the relative weight mechanism of the soleus muscle, and activates the muscle AR, and then activates the IGF-1/ IGF-1R-PI3K/Akt-m TOR signaling pathway. However, training passes through How AR mediates rapid muscle fiber content, MHC level and muscle glycogen content remains to be studied.
【學(xué)位授予單位】：上海體育學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：G804.2

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