A Study On The Multi - Target Of Delaying The Aging Of Yang Deficiency Mice

Objective:Preliminarily demonstrate the effect that Youguiwan can improve Yang-deficiency constitution of the mouse to delay aging. We can partly reveal the biological mechanism of the effect from the study on energy metabolism, repair of mitochondrion, expression levels of immunity and aging related genes and proteins in many organs. All our work is to provide a scientific basis for syndrome differentiation by constitution differentiation and for health maintenance and anti-aging.Methods:1. According to the principle of Xiao’s animal thermal behavior intelligent detection system, we design a simple animal thermal behavior model to choose mice of Yang-deficiency constitution. After mice treated by Youguiwan, we objectively evaluate the effect in comparison of the controlled group.2. Observe the difference of ultrastructure of mitochondria in the heart and brain among the 3 groups by electron microscope.3. Based on the previous research results, we use RT-PCR to test expression levels of aging-related genes, such as FHIT, HBD, MAP2K6, WNT5B, CSNK1D, FRAT2, DAPK2, GSK3A in the blood samples, and BDNF,TrKB in hippocampus among the three groups. We use in situ hybridization to test the expression levels of CaM in hypothalamus.4. We use immunohistochemistry method to measure expression levels of immunity and aging related proteins, such as FHIT, HBD, MAP2K6, WNT5B, CSNK1D, FRAT2, DAPK2, GSK3A, in the heart, spleen, kidney and head. We also test the expression levels of SOD and MDA in the blood.Results:1. After the treatment, fodder consumption rate and anal temperature rise in experimental group, while weight and rate of tending to 40 degree controlled plate step down. Compared with the Yang-deficiency controlled group, the fodder consumption rate, anal temperature, weight and rate of tending to 40 degree controlled plate show statistical significances, while survival time and visceral index of liver and thymus have a growing trend, visceral index of kidney, heart, lung, spleen, testis and adrenal gland show a decreasing tendency.2. The swelling of mitochondria and breakage of mitochondrial cristae in the heart and head tissues among the 3 groups gradually become worse in order of non-Yang-deficiency controlled group, Yang-deficiency experimental group and Yang-deficiency controlled group.3. Compared with Yang-deficiency controlled group, the genetic expression levels of CSNK1D, DAPK2 and WNT5B significantly lower, while BDNF, TrKB, HBD1 and CaM rise in the experimental group (P<0.05). Compared with the non-Yang-deficiency controlled group, expression levels of DAPK2 and WNT5B lower (P<0.01), while MAP2K6 and CaM rise (P<0.05).4. Compared with Yang-deficiency controlled group, proteic expression levels of WNT5B and FRAT2 in heart significantly lower in the experimental group, while HBD1 significantly rise, GSK3A WNT5B FRAT2 and DAPK2 significantly lower in spleen, WNT5B CSNK1D GSK3A significantly lower and MAP2K6 rise in kidney, WNT5B significantly lower, CSNK1D FRAT2 MAP2K6 and GSK3A significantly rise in head.Compared with non-Yang-deficiency controlled group, proteic expression levels of MAP2K6 significantly lower and DAPK2 significantly rise in spleen in the experimental group, while CSNK1D and DAPK2 significantly rise and MAP2K6 significantly lower in kidney, CSNK 1D and FRAT2 significantly rise and FHIT significantly lower in head.Conclusion:1. Preliminarily demonstrate the Preliminarily demonstrate the effect that Youguiwan can improve Yang-deficiency constitution to delay aging.2. Youguiwan can partly repair mitochondria ultrastructure to improve Yang-deficiency constitution to delay aging.3. It may be one of the biological effect mechanisms of improving Yang-deficiency constitution and delaying aging to suppress apoptosis and tumorigenesis, balance internal clock, enhance immune response and reduce harm of peroxide.4. Youguiwan may enhance the ability of nerve cell repairmen through improving Yang-deficiency constitution, which offer an experimental consult for Chinese herbs delaying aging and for clinical treatment of aging.