ABSTRACT
Since ancient times， delaying aging， health， and longevity have been the universal wish of people. Nowadays， China gives top strategic priority to the development of people's health. How to maintain a healthy life and slow down the aging of the human body is a problem worthy of our attention. Human aging can be shown as cell senescence from the microscopic level. Cell senescence is a process in which cell proliferation and differentiation and physiological function gradually decline. It is a normal physiological function responsible for the removal of damaged cells and is the regeneration and recovery of tissues after injury or acute stress. Aging is an irresistible natural law. Although it is inevitable， it is possible to delay aging. Energy metabolism is an important basis of cell function， in which cells use nutrients such as sugar and fat to produce adenosine triphosphate （ATP）. Mitochondria serve as the cell's power stations， where sugars， fats， and amino acids are eventually oxidized to release energy. Mitochondrial function decreases with age. Changes in mitochondrial dynamics， reactive oxygen species content， autophagy， and metabolites can cause dysfunction of electron transport chain and oxidative phosphorylation， and induce mitochondrial dysfunction. Mitochondrial dysfunction is one of the internal causes of many aging-related diseases， such as neurodegenerative diseases， Alzheimer′s disease， and atherosclerosis. Chinese medicine with few side effects and rich ingredients and health care moxibustion with safety and efficacy have been widely applied to the field of anti-aging. This study reviewed the effect of mitochondrial function on cell senescence， and retrieved， analyzed， and summarized research papers on the mechanism of traditional Chinese medicine （TCM） and moxibustion in delaying aging by affecting mitochondrial function， which is expected to provide new insights for further research in this field.
ABSTRACT
Objective To construct mouse dual specificity phosphatase-1 (DUSP1) 3'-untranslated region (3'-UTR) dual luciferase reporter system, and to identify its function. Methods The total RNA of RAW264.7 murine macrophage cells was extracted and reverse-transcribed into cDNA. The full length of mouse DUSP1 3'-UTR fragment was amplified by PCR and sub-cloned to the immediate downstream of luciferase cDNA in pGL3-control luciferase expression plasmid to obtain the recombinant plasmid pGL3-Luc-DUSP1- 3'UTR (LuDP). LuDP was then verified by PCR, restriction endonuclease and DNA sequencing analysis. Plasmid LuDP was transiently co-transfected into NIH3T3 cells with internal control plasmid pRL-TK, and the effects of DUSP1 3'-UTR on the expression of attached luciferase gene was analyzed with dual luciferase reporter assay system 48 hours after transfection. Both the luciferase expression module from LuDP and renilla luciferase expression module from pRL-TK were co-subcloned into pLenti6-TR vector to construct dual luciferase reporter plasmid LuDP/RL. The recombinant plasmid LuDP/RL was co-transfected with pcDNA3.1-HuR-FLAG (the eukaryotic expression plasmid of binding protein HuR) and miRNA mimics mmu-miR-101a into NIH3T3 cells respectively. The effects of HuR and mmu-mi-R101a on the expression of DUSP1 3'-UTR attached luciferase gene were also analyzed with dual luciferase reporter assay system. Results The dual luciferase reporter plasmid LuDP/RL was successfully constructed. The mouse DUSP1 mRNA 3'-UTR significantly down-regulated the expression of attached luciferase gene in NIH3T3 cells (0.14 ±0.01 fold, P<0.01). The co-transfected HuR up-regulated the expression of luciferase in LuDP/RL plasmid (1.40 ± 0.20 fold, P<0.05), and co-transfected mmu-miR-101a down-regulated the expression of LuDP/RL (0.57 ± 0.18 fold, P<0.05). Conclusion The constructed dual luciferase reporter plasmid LuDP/RL can be used to analyze the regulatory mechanism of mouse DUSP1 gene expression controlled by post-transcriptional regulatory elements.