| Cardiovascular aging refers to heart and vascular aging,which are the causative factors of a variety of acute and chronic diseases and have a huge impact on the physiology and psychology of the elderly.Therefore,it is vital to explore the research on delaying cardiovascular aging.In recent years,it has been found that mitochondria plays an important role in energy metabolism,biosynthesis and regulation of programmed cell death.As the age increases,the integrity of mitochondria and bioenergy efficiency all decline,resulting in a large number of aging-related disease phenotypes.Previous experiments showed that D-galactose can induce subacute aging animal model in a short time,and its mechanism and characteristics were similar to natural aging.Previous studies have shown that components of Yiqi Huoxue Yangyin Decoction can delay the aging process of heart and vessel in natural aging rats,mice and mice induced by high glucose and high fat.Therefore,by constructing a D-galactose-induced subacute aging mouse model,exploring the relationship between mitochondrial energy metabolism and cardiovascular aging,and exploring the effect of Yiqi Huoxue Yangyin Decoction by protecting the function of mitochondrial energy metabolism to delay the heart and aortic aging,which is of great significance.The first part:D-galactose induces aging heart and aorta in mice and the interventional effect of Yiqi Huoxue Yangyin DecoctionObjective:To build the subacute aging model of mice induced by D-galactose,observe the cardiovascular aging and the interventional effect of Yiqi Huoxue Yangyin Decoction.Methods:The sub-acute aging mice model was established by injecting D-galactose(180mg/kg)into the back of the neck,and the drug intervention was carried out for 10 weeks after 2 weeks.The experimental animals were divided into control group,aging model group,metformin group,low-dose group,medium-dose group and high-dose group.The whole experiment was carried out before sampling:open field test to observe the anxiety state and activity ability of mice,grip test to analyze the physical ability state of mice,laser speckle test to observe the microcirculation blood flow state of mice back.Superoxide dismutase and malondialdehyde levels in the serum were measured;HE staining was used to observe the morphology of the heart and aorta.and the intima-media thickness of the aorta was analyzed The expression of advanced glycationend products,matrix metalloprotein-2,tumor suppressor gene P53 and vascular endothelial-derived growth factor in the heart and aorta were detected by immunohistochemical staining.The relative telomere length and telomerase activity of heart and aortic tissues were measured.Results:(1)Compared with the blank control group,the aging model group:The body weight decreased before sampling(P<0.05);In the open field experiment,the times of standing increased(P<0.05),and the number of crossing grid increased significantly(P<0.01);In the laser speckle experiment,the blood perfusion was decreased(P<0.05);The activity of SOD was decreased(P<0.01);The content of MDA was increased(P<0.05);In heart tissue,the cardiomyocytes were arranged unevenly,myocardial fibers were broken,the expression of AGE protein was significantly increased(P<0.01),the expression of IMP-2,P53 protein were increased(P<0.05),and the relative length of telomeres is significantly shortened(P<0.01);In aortic tissue,it has abnormal morphology and structure,the medial intima was disordered and proliferated(P<0.05),the expression of AGE,MMP-2,and P53 protein was increased(P<0.05),and the expression of VEGF protein is significantly reduced(P<0.01).(2)Compared with the aging model group,the normalized grip strength(P<0.05),the volume of skin blood perfusion(P<0.01),the activity of SOD was increased(P<0.01)and the content of MDA was increased(P<0.05)in the middle dose of traditional Chinese medicine;In heart tissue,the shape of the it is relatively complete,the expression of AGE protein was down-regulated in all groups(P<0.05),MMP-2 protein was down-regulated in high and medium dose groups(P<0.01),and P53 protein was down-regulated in high and medium dose groups(P<0.05),the cardiac telomerase activity in metformin group was increased(P<0.05),the cardiac telomerase activity in low and middle dose groups was increased(P<0.01),and the telomere length in middle dose group was increased(P<0.01),the intimal thickness of aorta decreased in metformin group,low-dose group and middle-dose group(P<0.05),and the expression of AGE protein decreased in metformin group and middle-dose group(P<0.05),the expression of MMP-2 protein was down-regulated in low dose group(P<0.05),P53 protein was down-regulated in metformin group and middle dose group(P<0.05),the expression of VEGF protein was up-regulated in metformin group,middle-dose group and high-dose group(P<0.01).Conclusion:After D-galactose injection,the mice has been in a subacute aging state.Subacutely aging mice show signs of aging heart and aorta,which means that D-galactose can induce it.Yiqi Huoxue Yangyin Decoction can relief the aging heart and aorta of mice induced by D-galactose,and delay its development.The second section:Changes of Cardiovascular Energy Metabolism in Aging Mice and the Protective Effect of Yiqi Huoxue Yangyin DecoctionObjective:To observe the changes of energy metabolism in aging mice induced by D-galactose and the protective effect of Yiqi Huoxue Yangyin Decoction.Methods:The tissue samples of heart and aorta in D-galactose-induced cardiovascular aging mice were taken as the research objects.The fluorescence intensity of frozen sections of heart and aorta,the contents of Mitochondrial Ros(MTROS)and Mitochondrial membrane potential(MMP)were observed under Fluorescence microscope The activities of mitochondrial complexes Ⅰ,Ⅱ,Ⅲ,Ⅳ and Ⅴin mouse heart were detected by colorimetry and Elisa,and the expressions of ComplexⅠ,Ⅱ,Ⅲ,Ⅳ and V proteins in mouse aorta were detected by Western blot,to determine the damage of mitochondrial energy metabolism.Results:(1)Compared with the control group,the level of MTROS,MMP and the activity of mitochondrial complex Ⅰ were significantly increased(P<0.01),decreased(P<0.05)and decreased(P<0.01),the activities of complex Ⅱ,Complex Ⅲ,ComplexⅣ and Complex Ⅴ were significantly decreased(P<0.05),mTROS content increased(P<0.05),MMP decreased(P<0.05),protein expression of complex Ⅰ and complex Ⅳdecreased significantly(P<0.01).(2)compared with the aging model group,the contents of MTROS in the heart of mice in metformin group and high-dose group were significantly decreased(P<0.01),the contents of MTROS in middle-dose group were significantly decreased(P<0.05),MMP in metformin group and low-dose group were significantly increased(P<0.01),the activity of mitochondrial complex Ⅱ increased in metformin group and high dose group(P<0.05),and significantly increased in middle dose group(P<0.01),the activity of mitochondrial complexes Ⅲ was increased in metformin group,low-dose Chinese Medicine Group and middle-dose Chinese medicine group(P<0.05),and the activity of mitochondrial complexes IV was increased in metformin group and middle-dose Chinese medicine group(P<0.05),the contents of MTROS in metformin group and high-dose Chinese medicine group decreased(P<0.05),MMP in all Chinese medicine groups increased(P<0.05),the activity of mitochondrial complex I in metformin group significantly increased(P<0.01),and that of complex Ⅱ in metformin group significantly increased(P<0.05),the activity of mitochondrial complexes Ⅳ in metformin group and high-dose Chinese medicine group was significantly increased(P<0.01),and that of mitochondrial complexes V in metformin group,middle-dose Chinese Medicine Group and high-dose Chinese medicine group was significantly increased(P<0.01).Conclusion:D-galactose-induced aging mice have heart and aortic mitochondrial damage and decreased energy metabolism,which may be an important factor in cardiovascular aging;Yiqi Huoxue Yangyin Decoction can reduce mitochondrial ROS content and increase membrane potential.Improve the vitality of mitochondrial energy metabolism-related complexes,and delay the aging of the heart and aorta by protecting the mitochondrial energy metabolism function.Based on the results of the two parts of the experiment,it can be speculated that the dysfunction of mitochondrial energy metabolism may be closely related to cardiovascular aging.Yiqi Huoxue Yangyin Decoction may delay the aging of the heart and aorta by reducing the content of mtROS,increasing MMP,and protecting the activity of mitochondrial complex Ⅰ,Ⅱ,Ⅲ,Ⅳ,and Ⅴ. |
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