| Background:Chronic fatigue syndrome (CFS), which was officially named by the U.S. Centers for Disease Control and Prevention (CDC) in 1988, is an unexplained mental and physical sub-health status with duration more than 6 months. It often comes with fatigue, fever (or consciously fever), sore throat, muscle pain, joint pain, headaches, concentration, poor memory, sleep disorders and depression and other non -specific performance. With social lifestyle changing, environmental degradation and the impact of various factors in recent years, CFS onset showed an increasing trend in the number and the age of the incidence in young people is between the ages of 30-50 years old. Chronic fatigue syndrome is prevalent in modern populations, and it is an important risk factor for sudden death and premature aging. Chronic fatigue syndrome is called by "Consumption" in traditional Chinese medicine. Traditional Chinese medicine has a rich theory, long history and unique advantages in the prevention and treatment of disease. Our previous study found that Tonifying kidney Herbs have already displayed a better efficacy in the prevention and treatment of aging and age-related diseases. In the randomized, double-blind, placebo-controlled clinical trials, we found that tonifying kidney Herbs (Bushenyishou capsule etc.) could improve degenerative disease symptoms, inhibit T cell apoptosis and apoptosis-related gene transcription level in the elderly human beings and aged rats, etc. In a series of subsequent trials, we also found tonifying kidney Herbs can delay aging, old age related disease prevention by regulating the neuroendocrine immune network, improving immune function, antioxidant and free radical scavenging, etc. CFS pathogenesis and natural aging process has a similar path. May the tonifying kidney drug be used for prevention and treatment of CFS in young adults? And what are the therapeutic effects? we do not know yet. We will use Caenorhabditis elegans as CFS models in this study. The purpose is to explore high-throughput screening methods for the CFS therapy, drug evaluation index system for therapeutic effects, and preliminary mechanism of the drug action.Objective:1. To explore the high-throughput screening methods for CFS treatments by using Aβ1-42 transgenic C. elegans CFS model, and explore herbs in the effective dose;2. To evaluate the effects of 2,3,5,4’-tetrahydroxystilbene-2-O-β-D-glucoside(TSG) in improving the survival state of stress and the overall lifespan in C. elegans by simulating stress fatigue syndrome of various stress states.Methods:1. The L1-larvae were cultured with different concentrations of traditional Chinese herbs. In the control groups and herbs groups, internal Aβ protein toxicity (Aβ1-42 transgenic mutant CL4176) were exerted as the stresses for CFS, the ability of resisting stress was assayed by survival rate under stress.2. The L4-larvae were cultured with three concentrations of TSG:TSG -High (125μ M/L), TSG-Middle (62.5μM/L), TSG -Low (25μM/L). In the control group and TSG groups, internal Aβ protein toxicity (Aβ1-42 transgenic mutant CL4176), heat stress (35℃), and oxidative stress (juglone 250μg/mL) were exerted as he stresses, the ability of resisting stress was assayed by survival rate under stress. the survival vitality and related indicators analysis were assayed by overall average and maximum lifespan (25℃), movement distance in 30 seconds, quantitation of the number of GFP nuclei in PD4251 in control and TSG groups, pharyngeal pumping movement in 30 seconds and the egg number in reproductive period.Results:1. The high-throughput screening methods for CFS treatments was established by using Aβ1-42 transgenic C. elegans CFS model; Epimedium, Polygonum, two kinds of drugs which have therapeutic effects for C. elegans CFS model, were screened out of 17 kinds of traditional Chinese herbs.2. Compared to control group, in TSG-H and TSG-M group, the survival periods were significantly increased under Aβ protein toxicity (P<0.05),the average and maximum survival time in TSG groups were significantly increased under heat stress (P<0.05); in TSG-H group, the survival periods were significantly increased under oxidative stress (P<U.U5); The mean and maximum litespan were prolonged in TSG-H group (P<0.05). Compared to the control group, the active phase and average individual movement distance in TSG-H group were significantly extended (P<0.05); and pharyngeal pumping movement in BS-H group was higher (P<0.05). The degeneration of worms’body wall muscle with myo-3::GFP in PD4251 was also delayed by TSG-H treatment in Day4 and Day 10(25℃)(P<0.05). No significant difference was shown in spawning capability (P> 0.05) between the groups.Conclusions:1. The method of high-throughput sereening the herbs with Aβ1-42 transgenic C. elegans CFS model CL4176 was established that the water extract of herbs could be added directly to the NGM mediums, and Epimedium and Polygonum in the effective dose were the typical ones which enhance the stress capacity of C. elegans;2. TSG could significantly enhance the stress capacity of C. elegans against internal and external environment, and prolong their lifespan. It could also interfere their normal production, improve the quality of life, thus laying a foundation for further mechanism studies and pharmacological researches on TSG in preventing and treating CFS. |
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