The Creation Of Fatigue-type Sub-health Mice Model And The Effect Of Its Serum Corticosterone Caused By Weikang Granula

BACKGROUNDSub-health state is defined as no clear diagnosis, but a series of functional changes in body, is a middle state of human-health and illness, Its clinical manifestations include fatigue, decreased energy, decreased ability to respond and adapt to reduced capacity, Guangzhou (our group), Beijing and other relevant units of the sub-health epidemiological investigation showed that:sub-health population rate was 60% to 70%, and showed the characteristics of the following aspects:(1) universal; (2) individual consciousness is not easy; (3) the Objectivity and clarity of it are not recognized by modern medicine; (4) It can cause a variety of chronic diseases and can also be recovered to health. There are many clinical manifestations of sub-health state, but the large-scale epidemiological study by our group showed that fatigue is not only one of the important clinical manifestations of a sub-health state, but also contributed to the development of sub-health and fatigue-type sub-health is one of the main types of sub-health. Fast pace of life and work, a heavy psychological and social pressures, lack of rules of diet, adequate sleep and other factors may be related with the sub-health, but the ways in which these associated factors leading to fatigue-type sub-health state occurs is not clear. If the occurrence of fatigue-type sub-health related factors in classification, then, fatigue, chronic stress, anger, depression, sleep disorders can be categorized as physical or psychological stress areas. Stress is the body under stress at the time of the original stimulus non-specific systemic reactions occur, mainly through the release of a series of hormones and neurotransmitters to increase the body's basal metabolic level to mobilize the body in the best condition, to overcome damage to the original stress on the body, including adrenal cortical excitability increased glucocorticoid secretion, in the time of stress plays an important role. However, if the bad effects of stress on the body of the original continuous, the stress can be manifested as a dynamic and continuous process, by the corresponding target effects of hormones leading to disorder and disease occur within the environment. Fatigue, chronic stress, sleep disorders, stress is such a persistent source of bad stress, long-term effects on the body will result in "health-sub-health-illness" dynamic development, sub-health is between health and disease an intermediate between the state of health of its upstream and downstream migration of changes between the disease, this change is two-way migration, the formation of Health-Sub-health-the dynamic changes in the disease process, so early intervention to promote the health of its recovery and prevent the development of the disease is especially important. As the sub-health disease mechanism is not clear so there is not effective means of medical therapy. According to the factors of sub-health, Weikang-granula was invent to regulate the yin and yang, qi and blood of organs. According to the clinical observation shows that Weikang-granula has a good effect to reduce the fatigue of patients with sub-health. In order to further study the mechanisms of fatigue-type sub-health, it is necessary to introduce fatigue-type sub-health model to overcome the limitations of human trials, yet there is no relevant animal model. Therefore, this research is intended to establish the fatigue-type sub-health of mice, and research the mechanisms of fatigue-type sub-health under chronic fatigue stress base on mice model and research the mechanisms of the intervention of fatigue-type sub-health and the prevent form disease. This topic will be divided into two parts:PartⅠ:Establish fatigue-type sub-health mice model.PartⅡ:Preliminary study on the changes of fatigue-type sub-health mice's serum corticosterone caused by Weikang Granula.AIMPartⅠ:Simulate mainly sub-health factors, establish fatigue-type sub-health mice model. According to the characterize behavior, pathological and biochemical parameters and serum corticosterone levels of mice to define fatigue-type sub-health and evaluate the model.PartⅡ:To study the mechanism of action of relieving the fatigue of sun-health status by Weikang Granula and lay the foundation mechanism of targeted therapy.METHODSPartⅠ:Model group were based on fatigue-type sub-health rat model and improved to mice model. We kept mode mice in plastic boxes which size for 29cm x 18cm x 15cm and filled with water to a height of 0.8 cm to simulate the fatigue, crowded, lack of sleep, fear, anxiety and other stress factors to cause Sub-health state of fatigue. The normal control group does not offer any stimulation, free food. Weight-loaded forced swimming test was used to evaluate the extent of fatigue. Water temperature (25℃) The mice swam with a load of lead rings that weighed approximately 7% of their body weight and were attached to their tails. Exhaustive swimming standard was 10 seconds after mice sank into water without outcrop.Experiment 1:Determine the degree of the modeling Male healthy KM 40 mice in the SPF level was adopted in this study and randomly divided into 6h/d,8h/d, 10h/d model group and normal control group(n=10). Once every 3 days to evaluate fatigue by exhaustive swimming time in the period of 21 days.Experiment 2:Determine the days of making mode Male healthy KM 150 mice in the SPF level was adopted in this study and randomly divided into control and mode groups, each of which contained 5 groups (0,5,9,12, 16 days groups, n=10). Mode mice were kept in plastic boxes 8h/day. Weight-loaded forced swimming test was used to evaluate the extent of fatigue before the mice would be killed. The method of radioimmunity was used to test the level of Serum corticosterone.Experiment 3:the evaluation of model Male healthy KM 40 mice in the SPF level were randomly divided into normal control group and the fatigue of sub-health model group (8h/day for 9 days of mould), natural restoration for 7 days group (rest 7 days after modeling), natural restoration for 14 days (rest 14 days after modeling), each group has 10 mice. Weight-loaded forced swimming test was used to evaluate the extent of fatigue before the mice would be killed. The method of radioimmunity was used to test the level of Serum corticosterone.PartⅡ:Fatigue-type Sub-health model group, natural restoration for 7 days group(normal saline for 7 days after 9 days modeling), weikang high, medium and low dose group were given high, medium and low doses of Weikang Granula solution orally for 7 days after 9 days modeling. Normal control group were given nothing stimulate.Experiment 4:Male healthy KM 60 mice in the SPF level were randomly divided into normal control group, fatigue-type Sub-health model group, natural restoration for 7 days group, weikang high, medium and low dose group. Weight-loaded forced swimming test was used to evaluate the extent of fatigue before the mice would be killed. The method of radioimmunity was used to test the level of Serum corticosterone.RESULTSExperiment 1:With the longitudinal extension of modeling time:there was significant decreased (P<0.01) in swimming time at the fist 12 days in modeling 6h group. There was significant difference (P<0.05) in swimming time at the first 3 days in modeling 8h group.6-12 days decreased significantly. There was significant difference (P<0.01) in swimming time at the first 3 days in modeling 10h group.Experiment 2:With the longitudinal extension of experiment time:(1) There was significant increased (P<0.05) in the weight of body of the control group, but on significant difference(P> 0.05) in model group.(2)There was no significant difference(P> 0.05) in swimming time in control group, but gradually decreased in model group, There was significant decreased in 5 day group. (3) Serum levels of corticosterone in mice modeling 9 days ago there was no significant difference (P> 0.05),9 days later was significantly lower (P<0.05). (4) modeling 9 days, serum biochemistry, blood and the normal control group showed no significant difference (P> 0.05).Experiment 3:Compared With the normal control group:(1) The swimming time and serum corticosterone of fatigue-type sub-health model group were significantly different (P<0.05). (2) Recovery in 7 days group's swimming time was significantly different (P<0.05), while serum corticosterone was no significant differences (P> 0.05). (3) 14-day recovery group's swimming time and serum corticosterone were not significantly different (P>0.05). (4) There were significant increase in swimming time (P<0.05) and significantly lower in serum corticosterone (P<0.05) between recovery for 7days and 14 days.Experiment 4:Compared with the normal control group:fatigue-type sub-health model group swimming time was significantly lower (P<0.05), serum corticosterone increased significantly (P<0.05), The swimming time and blood corticosterone of Weikang Granula high dose group and medium dose group have no significant difference (P> 0.05), Weikang Granula low dose group was significantly lower in swimming time (P<0.05), significantly increased in serum corticosterone (P<0.05).CONCLUSION:1) We successfully created mouse model of fatigue-type sub-health by simulating the occurrence of fatigue-type sub-health related factors, continue to act in mice 9 days,8 hours a day.2) Our studies show that if fatigue-type sub-health risk factors were removed, sub-health state can be recovered by themselves, but need a longer time, intervention by the Weikang Granula can relieve fatigue in mice, block the development to the disease, reduce Sub-health recovery time.3) Fatigue, sleep disturbance, psychological stress factors such as continuing the role of chronic stress caused by adrenal pathological hyperactivity, glucocorticoid may be involved in the pathological increase in the incidence of fatigue-type sub-health development.4) Weikang Granula can lower the level of serum corticosterone of the fatigue-type sub-health, which may be one of the pharmacological effects to the fatigue-type sub-health.