【摘要】：研究目的:通過建立中老年大鼠8周恒定負(fù)荷有氧運動以及姜黃素干預(yù)模型,檢測運動及姜黃素干預(yù)后大鼠比目魚肌AMPK活性變化以及自噬相關(guān)基因的表達(dá)情況,并運用HE染色去觀察大鼠骨骼肌的組織病理學(xué)變化,探討運動結(jié)合姜黃素誘導(dǎo)的AMPK活性變化對中老年大鼠骨骼肌細(xì)胞自噬的影響。推測運動和姜黃素預(yù)防Sarcopenia的作用機理。研究方法:同批次健康雄性21月齡SD大鼠40只,SPF(Specific pathogen Free,SPF)級動物,分籠飼養(yǎng)。適應(yīng)性喂養(yǎng)三天,隨機分為4組,隨機分為空白對照組(BC,n=10)、安靜姜黃素組(QC,n=10)、恒定負(fù)荷運動+姜黃素組(EC,n=10),恒定負(fù)荷運動(NE,n=10)。每組分2籠喂養(yǎng),每籠5只。運動組大鼠采用跑臺運動方式,建立姜黃素結(jié)合有氧運動訓(xùn)練干預(yù)方式動物實驗?zāi)Ｐ?其中坡度為0,速度15m/min×60min(負(fù)荷強度相當(dāng)于70%VO2max),姜黃素劑量為100mg/kg老鼠體重,每周持續(xù)訓(xùn)練5天,共訓(xùn)練8周。訓(xùn)練末次結(jié)束后20小時,完整取下大鼠后肢比目魚肌,計算大鼠比目魚肌的相對重量(SI),采用Elisa法檢測比目魚肌AMPK活性變化,通過實時熒光定量PCR檢測AGT7 m RNA、LC3-b mRNA、LAMP-2mRNA的表達(dá)情況,并從形態(tài)學(xué)方面運用HE染色觀察大鼠骨骼肌的組織病理學(xué)變化。研究結(jié)果:1)各組大鼠體重均顯著降低(p0.01);實驗后,qc組、ne組及ec大鼠體重均顯著高于bc組(p0.01),ne組高于qc組和ec組(p0.05)。2)與bc組相比較,qc組、ne組以及ec組的si值都要高,且具有非常顯著性差異(p0.01);ne組比ec組要高,且具有顯著性差異(p0.05)。3)各組中老年大鼠比目魚肌出現(xiàn)了不同程度的與衰老相關(guān)的組織病理學(xué)變化,相對于bc組,qc組、ne組以及ec組的織異常像出現(xiàn)的頻數(shù)較低,其中ne組表現(xiàn)最好。4)與bc組相比,qc組、ne組及ec組ampk活性明顯提高(p0.01);其中,ec組比目魚肌ampk活性最高,與ne組、qc組相比,都具有顯著性差異(p0.05)。5)與對照組bc組相比,ne組和ec組agt7mrna的表達(dá)水平明顯要高(p0.01),qc組agt7mrna的表達(dá)水平高于bc組(p0.05),ne與ec組、qc組相比明顯提高(p0.01)。6)與bc組相比,qc組、ne組以及ec組lc3-bmrna表達(dá)水平明顯增高(p0.01),ec組lc3-bmrna表達(dá)水平明顯高于qc組和ne組(p0.01)。7)ec組、qc組以及ne組lamp-2mrna的表達(dá)量明顯高于bc組(p0.01),且ec組要高于qc組和ne組p0.05),ne組要高于qc組(p0.05)。研究結(jié)論:1)8周的恒定負(fù)荷有氧運動、姜黃素及兩者聯(lián)合干預(yù)都能在一定程度上維持骨骼肌重量,改善骨骼肌組織的病變,其機理可能是通過激活骨骼肌AMPK,提高中老年大鼠骨骼肌AGT7、LC3-b、LAMP-2基因的表達(dá)來實現(xiàn)。2)姜黃素和恒定負(fù)荷有氧運動可能是通過激活骨骼肌AMPK去誘導(dǎo)自噬這一機制來維持骨骼肌質(zhì)量及改善骨骼肌組織病變。3)8周恒定負(fù)荷有氧運動比起姜黃素和姜黃素聯(lián)合恒定負(fù)荷有氧運動對于緩解骨骼肌質(zhì)量流失及改善肌組織病變的作用更明顯,可能是由于恒定負(fù)荷有氧運動能使骨骼肌維持著一種更適應(yīng)的自噬水平。
[Abstract]:Objective: to study the changes of AMPK activity in soleus muscle and the expression of autophagy related genes in soleus muscle after exercise and curcumin intervention by establishing 8 weeks constant load aerobic exercise and curcumin intervention model of middle-aged and aged rats. HE staining was used to observe the histopathological changes of skeletal muscle in rats and to explore the effect of exercise combined with curcumin on the autophagy of skeletal muscle cells induced by curcumin. The mechanism of exercise and curcumin in preventing Sarcopenia was speculated. Methods: 40 healthy male SD rats of 21 months old were raised in cage. After three days of adaptive feeding, they were randomly divided into four groups: blank control group (BC,n=10), quiet curcumin group (QC,n=10), constant load exercise group (EC,n=10) and constant load exercise group (NE,n=10). Each group was fed with 2 cages, 5 in each cage. In the exercise group, the animal model of curcumin combined with aerobic exercise intervention was established, in which the slope was 0, the speed of 15m/min 脳 60min (load intensity was equal to 70%VO2max), and the dose of curcumin was the weight of 100mg/kg mice, which was continuously trained for 5 days per week. A total of 8 weeks of training. 20 hours after the last training, the soleus muscle of the hind limb of the rats was removed completely, and the relative weight of the soleus muscle was calculated. The AMPK activity of the soleus muscle was detected by Elisa method. The expression of AGT7 m RNA,LC3-b mRNA,LAMP-2mRNA was detected by real-time fluorescence quantitative PCR. The histopathological changes of rat skeletal muscle were observed by HE staining. The results showed that the weight of rats in each group was significantly decreased (p0.01), and the weight of ec rats in QC group was significantly higher than that in qc group (p0.01) and ec group (p0.05) .2) compared with bc group, the si values of QC group and ec group were significantly higher than those of bc group (p0.01) and ec group (p0.05), and compared with bc group, the si values of QC group and ec group were significantly higher than those of control group (p0.01). There were significant differences (p0.01) in the soleus muscle between the two groups (p0.01) and the histopathological changes related to aging in the soleus muscle of the middle-aged and aged rats were higher than those in the ec group (p0.05). Compared with the bc group, the frequency of abnormal tissue appearance in the ne group was lower than that in the bc group (P 0.01), and the activity of ampk in the bc group was significantly higher than that in the bc group (p0.01), and the ampk activity of the flounder muscle in the ne group was the highest, which was higher than that in the ne group (P < 0.01), and the activity of ampk in the flounder muscle was the highest in the ne group compared with the bc group (P < 0.05). There was significant difference (p0.05) between the two groups (p0.05). Compared with the control group, the expression of agt7mrna in the ne group and the ec group was significantly higher than that in the bc group (p0.05) and the agt7mrna expression level in the bc group was significantly higher than that in the bc group (p0.05), compared with the ec group, the agt7mrna expression level in the ec group was significantly higher than that in the control group (p0.01). Compared with the bc group, the agt7mrna expression level in the ne group and the ec group was significantly higher than that in the control group (p0.05). The expression of lc3-bmrna was significantly higher in the lc3-bmrna group than in the qc group and the ne group (p0.01). The expression of lamp-2mrna in the ec group was significantly higher than that in the bc group (p0.01), and the expression of lamp-2mrna in the ec group was higher than that in the qc group and ne group (p0.05). The expression of lamp-2mrna in the ec group was significantly higher than that in the qc group (p0.05). Conclusion: 8 weeks of constant load aerobic exercise, curcumin and combined intervention of curcumin and both can maintain skeletal muscle weight to a certain extent and improve the pathological changes of skeletal muscle tissue. The mechanism may be to increase the expression of AGT7,LC3-b,LAMP-2 gene in skeletal muscle of middle-aged and aged rats by activating skeletal muscle AMPK,.) curcumin and constant load aerobic exercise may maintain bone by activating skeletal muscle AMPK to induce autophagy. Compared with curcumin and curcumin combined with constant load aerobic exercise, the effects of 8 weeks of constant load aerobic exercise on alleviating the loss of skeletal muscle mass and improving the pathological changes of skeletal muscle were more obvious than those of curcumin and curcumin combined with constant load aerobic exercise. This may be because constant-load aerobic exercise keeps skeletal muscles at a more adaptive autophagy level.
【學(xué)位授予單位】：湖南師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：G804.2

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