姜黄素和有氧运动诱导的AMPK激活对中老年大鼠骨骼肌细胞自噬的影响初探

发布时间：2018-09-18 17:27

【摘要】：研究目的:通过建立中老年大鼠8周恒定负荷有氧运动以及姜黄素干预模型,检测运动及姜黄素干预后大鼠比目鱼肌AMPK活性变化以及自噬相关基因的表达情况,并运用HE染色去观察大鼠骨骼肌的组织病理学变化,探讨运动结合姜黄素诱导的AMPK活性变化对中老年大鼠骨骼肌细胞自噬的影响。推测运动和姜黄素预防Sarcopenia的作用机理。研究方法:同批次健康雄性21月龄SD大鼠40只,SPF(Specific pathogen Free,SPF)级动物,分笼饲养。适应性喂养三天,随机分为4组,随机分为空白对照组(BC,n=10)、安静姜黄素组(QC,n=10)、恒定负荷运动+姜黄素组(EC,n=10),恒定负荷运动(NE,n=10)。每组分2笼喂养,每笼5只。运动组大鼠采用跑台运动方式,建立姜黄素结合有氧运动训练干预方式动物实验模型,其中坡度为0,速度15m/min×60min(负荷强度相当于70%VO2max),姜黄素剂量为100mg/kg老鼠体重,每周持续训练5天,共训练8周。训练末次结束后20小时,完整取下大鼠后肢比目鱼肌,计算大鼠比目鱼肌的相对重量(SI),采用Elisa法检测比目鱼肌AMPK活性变化,通过实时荧光定量PCR检测AGT7 m RNA、LC3-b mRNA、LAMP-2mRNA的表达情况,并从形态学方面运用HE染色观察大鼠骨骼肌的组织病理学变化。研究结果:1)各组大鼠体重均显著降低(p0.01);实验后,qc组、ne组及ec大鼠体重均显著高于bc组(p0.01),ne组高于qc组和ec组(p0.05)。2)与bc组相比较,qc组、ne组以及ec组的si值都要高,且具有非常显著性差异(p0.01);ne组比ec组要高,且具有显著性差异(p0.05)。3)各组中老年大鼠比目鱼肌出现了不同程度的与衰老相关的组织病理学变化,相对于bc组,qc组、ne组以及ec组的织异常像出现的频数较低,其中ne组表现最好。4)与bc组相比,qc组、ne组及ec组ampk活性明显提高(p0.01);其中,ec组比目鱼肌ampk活性最高,与ne组、qc组相比,都具有显著性差异(p0.05)。5)与对照组bc组相比,ne组和ec组agt7mrna的表达水平明显要高(p0.01),qc组agt7mrna的表达水平高于bc组(p0.05),ne与ec组、qc组相比明显提高(p0.01)。6)与bc组相比,qc组、ne组以及ec组lc3-bmrna表达水平明显增高(p0.01),ec组lc3-bmrna表达水平明显高于qc组和ne组(p0.01)。7)ec组、qc组以及ne组lamp-2mrna的表达量明显高于bc组(p0.01),且ec组要高于qc组和ne组p0.05),ne组要高于qc组(p0.05)。研究结论:1)8周的恒定负荷有氧运动、姜黄素及两者联合干预都能在一定程度上维持骨骼肌重量,改善骨骼肌组织的病变,其机理可能是通过激活骨骼肌AMPK,提高中老年大鼠骨骼肌AGT7、LC3-b、LAMP-2基因的表达来实现。2)姜黄素和恒定负荷有氧运动可能是通过激活骨骼肌AMPK去诱导自噬这一机制来维持骨骼肌质量及改善骨骼肌组织病变。3)8周恒定负荷有氧运动比起姜黄素和姜黄素联合恒定负荷有氧运动对于缓解骨骼肌质量流失及改善肌组织病变的作用更明显,可能是由于恒定负荷有氧运动能使骨骼肌维持着一种更适应的自噬水平。
[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.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：G804.2

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