藥膳加貼敷對衰老大鼠骨骼肌抗氧化能力及細胞凋亡影響的實驗研究
發(fā)布時間：2019-03-21 17:12
【摘要】：目的:本實驗基于“脾主肌肉四肢”理論,選用D-半乳糖(D-a lactose,D-gal)致衰老大鼠為研究對象,采用補脾益氣藥膳灌胃加穴位貼敷的療法對衰老模型大鼠進行干預,觀察衰老大鼠骨骼肌形態(tài)學改變、抗氧化能力及cas pase-3和caspase-8的表達影響,了解骨骼肌質量的漸進性下降與細胞凋亡和抗氧化能力之間的內(nèi)在聯(lián)系和作用機制。為延緩肌肉衰減綜合征(Sarcopenia)的發(fā)生提供理論依據(jù),并最終為臨床干預Sarcopenia提供有效的護理措施,以便提高老年人日�；顒幽芰Α⒆岳砟芰�,減少跌倒及臥床的機率,改善老年人生活質量,降低醫(yī)療保健成本及家庭和社會的經(jīng)濟負擔。材料與方法:選取SPF(Specific pathogen Free,SPF)級3月齡健康雄性Spr ague-DaWley(SD)大鼠36只,隨機分為空白組、模型空白組和藥膳貼敷療法組,每組12只。模型空白組和藥膳貼敷組每日腹部皮下注射7.5%D-gal 125mg/kg,空白組每日腹部皮下注射0.9%生理鹽水125mg/kg,造模持續(xù)6周。6周后,藥膳貼敷組給與藥膳灌胃(10ml·kg-1·d-1)和穴位貼敷干預,空白組和模型空白組給與等劑量的生理鹽水和空白貼敷干預,干預持續(xù)4周。最后一次灌胃24小時后,進行腹腔麻醉,經(jīng)腹主動脈取血,迅速剝離股直肌。通過HE染色鏡下觀察衰老大鼠股直肌細胞形態(tài)結構的變化,PCR檢測股直肌含半胱氨酸的天冬氨酸蛋白水解酶3(Cysteinyl aspa rtate specifi c proteinase-3,Caspase-3)和Caspase-8mRNA的表達,ELISA檢測血清中谷胱甘肽過氧化物酶(glutathione peroxidase,GSH-Px)、超氧化物歧化酶(superoxide dismutase,SOD)的活性和丙二醛(Mal ondialdehyde,MDA)的含量及股直肌GSH-Px、SOD、Cas pase-3 Caspase-8的活性和MDA的含量。采用SPSS17.0統(tǒng)計分析軟件和Microsoft Office Excel 2007進行數(shù)據(jù)處理分析,每組數(shù)據(jù)用平均數(shù)±標準差(Mean±S)表示,組間差異采用單因素方差分析,差異顯著性水平定義為P0.05。結果:1.模型空白組和藥膳貼敷組大鼠都有衰老的體貌特征,但藥膳貼敷組大鼠較模型空白組大鼠衰老特征有所減輕;三組大鼠體重均有增加,模型空白組大鼠略低于空白組,但組間比較無顯著性差異(P0.05)。2.光鏡下:空白組大鼠為正常骨骼肌組織學特征,模型空白組和藥膳貼敷組大鼠都出現(xiàn)了不同程度的肌細胞萎縮或退行性改變,而藥膳貼敷組大鼠肌細胞的萎縮程度比模型空白組較輕。3.血清學指標:模型空白組大鼠血清中GSH-Px、SOD的活性較空白組顯著下降(P0.05),而MDA的含量較空白組顯著增加(P0.05);與模型空白組相比,藥膳貼敷組大鼠血清中GSH-Px、SOD的活性顯著提高(P0.05),而MDA的含量顯著降低(P0.05)。4.骨骼肌組織生化指標:與空白組相比,模型空白組大鼠骨骼肌中GSH-Px、SOD的活性顯著下降(P0.05),MDA的含量及Caspase-3、Caspase-8的活性顯著提高(P0.05);與模型空白組相比,藥膳貼敷組大鼠骨骼肌中GSH-Px、SOD顯著提高(P0.05),MDA的含量及Caspase-3、Caspase-8的活性顯著下降(P0.05)。5.骨骼肌組織分子生物學指標:與空白組相比,模型空白組大鼠骨骼肌中Caspase-3、Caspase-8mRNA表達明顯增加(P0.05);與模型空白組相比,藥膳貼敷組大鼠模型組大鼠骨骼肌中Caspase-3、Caspase-8mRNA(P0.05)表達明顯減少。結論:1.藥膳加穴位貼敷療法能夠改善老年大鼠的體貌、狀態(tài)及骨骼肌組織形態(tài)學特征,并且能夠減緩肌肉衰減綜合征的發(fā)生及發(fā)展。2.藥膳加穴位貼敷療法能夠提高衰老大鼠血清及骨骼肌中GSH-Px、SOD的活性,降低MDA的含量,從而提高骨骼肌的抗氧化能力,減輕骨骼肌的氧化損傷,延緩骨骼肌的衰老。3.藥膳加穴位貼敷療法能夠降低衰老大鼠骨骼肌細胞中Caspase-3和Caspase-8的活性,下調Caspase-3mRNA和Caspase-8mRN A的表達,進而減少細胞凋亡,延緩骨骼肌的衰老。
[Abstract]:Objective: To study the changes of skeletal muscle morphology in aging rats by using D-galactose (D-gal)-induced aging rats to study the aging model rats based on the "spleen main muscle and limbs" theory. The relationship between the progressive decline of skeletal muscle mass and the cell apoptosis and the anti-oxidation ability was studied by the effect of the anti-oxidation ability and the expression of caspase-8. The invention provides a theoretical basis for delaying the occurrence of the muscle attenuation syndrome (Sarcoenia), and finally provides effective nursing measures for the clinical intervention Sarcoenia, so as to improve the daily activity capability of the old people, self-adjustment, reduce the probability of falling and bedridden, improve the quality of life of the old people, And the economic burden of the health care cost and the family and the society is reduced. Materials and Methods:36 healthy male Sprague-DaWley (SD) rats were randomly divided into blank group, model blank group and medicated diet therapy group. The daily abdominal subcutaneous injection of the model blank group and the medicated diet group was 7.5% D-gal 125 mg/ kg. The daily abdomen of the blank group was injected with 0.9% normal saline of 125 mg/ kg and the model was established for 6 weeks. After 6 weeks, the medicated diet was administered to the medicated diet (10 ml 路 kg-1 路 d-1) and the acupoint application for intervention. The blank group and the model blank group were given equal dose of normal saline and blank for intervention, and the intervention lasted for 4 weeks. After the last administration for 24 hours, the abdominal cavity was anesthetized, the blood was taken from the abdominal aorta, and the rectus femoris was quickly separated. The changes of the morphological structure of the straight muscle cells of the aging rats were observed by HE staining, and the expression of cysteinyl aspa rate specific c protein-3 (Caspase-3) and Caspase-8 mRNA was detected by PCR, and the glutathione peroxidase (GSH-Px) in the serum was detected by ELISA. The activity of superoxide dismutase (SOD) and the content of MDA and the activity of GSH-Px, SOD, Cas ase-8 and the content of MDA were studied. The data was analyzed by SPSS17.0 statistical analysis software and Microsoft Office Excel 2007. The mean standard deviation (Mean-S) of each group of data indicated that the difference between the groups was defined as P0.05 with a single-factor analysis of variance. Results:1. In the model blank group and the medicated diet group, the aging characteristics of the rats in the model blank group and the medicated diet group were reduced; the weight of the three groups of rats was increased, and the rats in the blank group of the model group were slightly lower than that of the blank group, but no significant difference was found between the groups (P0.05). Under the light microscope, in the blank group, the histological characteristics of the normal skeletal muscle, the blank group of the model group and the rats of the medicated diet group had different degrees of myocyte atrophy or degeneration, and the extent of the atrophy of the myoblasts in the medicated diet group was less than that of the blank group of the model. The results showed that the activity of GSH-Px and SOD in the serum of the model blank group was significantly lower than that in the blank group (P0.05). Compared with the blank group of the model, the activity of GSH-Px and SOD in the serum of the medicated diet group was significantly increased (P0.05). The content of MDA was significantly lower (P0.05). Compared with the blank group, the activity of GSH-Px and SOD in the skeletal muscle of the model blank group decreased significantly (P0.05). Compared with the blank group of the model, the activity of GSH-Px in the skeletal muscle of the medicated diet group was higher than that of the blank group (P0.05). The content of MDA and the activity of Caspase-3 and Caspase-8 decreased significantly (P0.05). Compared with the blank group, the expression of Caspase-3 and Caspase-8 mRNA in the skeletal muscle of the model group was significantly lower than that of the blank group (P0.05). Conclusion:1. The medicated diet plus acupoint application can improve the physical appearance, state and skeletal muscle tissue morphology of the old rats, and can slow down the occurrence and development of the muscle attenuation syndrome. The medicated diet plus acupoint application can improve the activity of GSH-Px and SOD in the serum and skeletal muscle of the aging rats, and decrease the content of MDA, so as to improve the antioxidant capacity of the skeletal muscle, reduce the oxidative damage of the skeletal muscle and delay the aging of the skeletal muscle. It is possible to reduce the activity of Caspase-3 and Caspase-8 in the skeletal muscle cells of aging rats, and to reduce the expression of Caspase-3 mRNA and Caspase-8 mRN A, so as to reduce the cell apoptosis and delay the aging of the skeletal muscle.
【學位授予單位】：遼寧中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R248
本文編號：2445141
[Abstract]:Objective: To study the changes of skeletal muscle morphology in aging rats by using D-galactose (D-gal)-induced aging rats to study the aging model rats based on the "spleen main muscle and limbs" theory. The relationship between the progressive decline of skeletal muscle mass and the cell apoptosis and the anti-oxidation ability was studied by the effect of the anti-oxidation ability and the expression of caspase-8. The invention provides a theoretical basis for delaying the occurrence of the muscle attenuation syndrome (Sarcoenia), and finally provides effective nursing measures for the clinical intervention Sarcoenia, so as to improve the daily activity capability of the old people, self-adjustment, reduce the probability of falling and bedridden, improve the quality of life of the old people, And the economic burden of the health care cost and the family and the society is reduced. Materials and Methods:36 healthy male Sprague-DaWley (SD) rats were randomly divided into blank group, model blank group and medicated diet therapy group. The daily abdominal subcutaneous injection of the model blank group and the medicated diet group was 7.5% D-gal 125 mg/ kg. The daily abdomen of the blank group was injected with 0.9% normal saline of 125 mg/ kg and the model was established for 6 weeks. After 6 weeks, the medicated diet was administered to the medicated diet (10 ml 路 kg-1 路 d-1) and the acupoint application for intervention. The blank group and the model blank group were given equal dose of normal saline and blank for intervention, and the intervention lasted for 4 weeks. After the last administration for 24 hours, the abdominal cavity was anesthetized, the blood was taken from the abdominal aorta, and the rectus femoris was quickly separated. The changes of the morphological structure of the straight muscle cells of the aging rats were observed by HE staining, and the expression of cysteinyl aspa rate specific c protein-3 (Caspase-3) and Caspase-8 mRNA was detected by PCR, and the glutathione peroxidase (GSH-Px) in the serum was detected by ELISA. The activity of superoxide dismutase (SOD) and the content of MDA and the activity of GSH-Px, SOD, Cas ase-8 and the content of MDA were studied. The data was analyzed by SPSS17.0 statistical analysis software and Microsoft Office Excel 2007. The mean standard deviation (Mean-S) of each group of data indicated that the difference between the groups was defined as P0.05 with a single-factor analysis of variance. Results:1. In the model blank group and the medicated diet group, the aging characteristics of the rats in the model blank group and the medicated diet group were reduced; the weight of the three groups of rats was increased, and the rats in the blank group of the model group were slightly lower than that of the blank group, but no significant difference was found between the groups (P0.05). Under the light microscope, in the blank group, the histological characteristics of the normal skeletal muscle, the blank group of the model group and the rats of the medicated diet group had different degrees of myocyte atrophy or degeneration, and the extent of the atrophy of the myoblasts in the medicated diet group was less than that of the blank group of the model. The results showed that the activity of GSH-Px and SOD in the serum of the model blank group was significantly lower than that in the blank group (P0.05). Compared with the blank group of the model, the activity of GSH-Px and SOD in the serum of the medicated diet group was significantly increased (P0.05). The content of MDA was significantly lower (P0.05). Compared with the blank group, the activity of GSH-Px and SOD in the skeletal muscle of the model blank group decreased significantly (P0.05). Compared with the blank group of the model, the activity of GSH-Px in the skeletal muscle of the medicated diet group was higher than that of the blank group (P0.05). The content of MDA and the activity of Caspase-3 and Caspase-8 decreased significantly (P0.05). Compared with the blank group, the expression of Caspase-3 and Caspase-8 mRNA in the skeletal muscle of the model group was significantly lower than that of the blank group (P0.05). Conclusion:1. The medicated diet plus acupoint application can improve the physical appearance, state and skeletal muscle tissue morphology of the old rats, and can slow down the occurrence and development of the muscle attenuation syndrome. The medicated diet plus acupoint application can improve the activity of GSH-Px and SOD in the serum and skeletal muscle of the aging rats, and decrease the content of MDA, so as to improve the antioxidant capacity of the skeletal muscle, reduce the oxidative damage of the skeletal muscle and delay the aging of the skeletal muscle. It is possible to reduce the activity of Caspase-3 and Caspase-8 in the skeletal muscle cells of aging rats, and to reduce the expression of Caspase-3 mRNA and Caspase-8 mRN A, so as to reduce the cell apoptosis and delay the aging of the skeletal muscle.
【學位授予單位】：遼寧中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R248
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