Effect Of Large Plant Rhodiola Capsule On High Altitude Hypoxia-induced Fatigue,mood And Cognitive Impairment

The vast plateau area of our country not only has an important economic and political position,but also an important military strategic position.It is the key to the country’s social stability.However,the high altitude hypoxic environment has an important impact on the body’s exercise and cognitive functions,resulting in a decline in the group of people exposed to hypoxia,anxiety and depression,and a decline in cognitive function,which seriously threatens the military operation efficiency of the officers and soldiers of the plateau troops.Therefore,it is of great significance to find a drug that can not only enhance the exercise tolerance of plateau,but also help to improve the emotional and cognitive impairment caused by hypoxia exposure in plateau to improve the combat effectiveness and operational ability of plateau troops.Large plant rhodiola capsule are prepared from the water extract of the dried roots of Rhodiola rosea.They are clinically used for the treatment of cardiovascular diseases such as coronary heart disease and angina pectoris.The impact of dysfunction has not yet been studied.The compound Danshen dripping pill is an anti-hypoxic drug equipped by our army.It is a traditional Chinese medicine for the treatment of cardiovascular diseases that has completed the second clinical trial in the United States.The study first evaluated the effects of Large plant rhodiola capsule on the ability to withstand atmospheric hypoxia and anti-fatigue in mice,and then simulated acute altitude hypoxia exposure to observe the changes in fatigue,anxiety,depression and cognitive function in mice.On the basis of observing the effects of Large plant rhodiola capsule on fatigue,mood and cognitive impairment caused by simulated acute altitude hypoxia exposure,and finally discussing how Large plant rhodiola capsule can improve anxiety,depression and cognitive impairment caused by altitude hypoxia In order to reveal the potential application value of Large plant rhodiola capsule in improving brain-body dysfunction related to high altitude hypoxia.Chapter 1 Improve normobaric hypoxia tolerance and anti-fatigue effect of large plant Rhodiola capsule in miceThe male BALB/c mice aged 6～8 weeks were randomly divided into control,compound Danshen dripping pill(CDDP,0.122 g·kg^-1),and large plant Rhodiola capsule(LPRC,0.35,0.7,and 1.4 g·kg^-1)group（n=10）.Mice were ig administered with saline,CDDP,or LPRC for seven days,respectively.Anoxia endurance of LPRC was assessed by a gas mixture of low oxygen test and hypoxia in hermetic space test.The anti-fatigue effect of LPRC was evaluated by a weight-loaded swimming test and rotarod test.Colorimetric method was used to detect the changes of glycogen content in liver and gastrocnemius tissue,fatigue-related indexes[including the content of urea nitrogen（BUN）and lactic acid（BLA）,the activity of lactate dehydrogenase（LDH）]and oxidative stress-related indexes[including the activity of superoxide dismutase（SOD）,glutathione peroxidase（GSH-PX）,the content of malondialdehyde（MDA）]in serum in exhaustive mice after non-load swimming.1.Improve normobaric hypoxia tolerance and anti-fatigue effect of large plant Rhodiola capsule in miceThe hypoxia survival time and weight-loaded swimming time of mice in the LPRC(0.7,1.4 g·kg^-1)were significantly longer than the control group.The time in rotarod of learning phase and test phase of mice in the LPRC 1.4g·kg^-1was significantly longer than the control group.2.Effects of Large plant Rhodiola Capsule on Related Indexes of Fatigue in Exhausted Swimming MiceAfter non-load swimming,the contents of liver glycogen and muscle glycogen and the activities of LDH,SOD,and GSH-PX in the LPRC were significantly higher than the control group.Simultaneously,BUN,BLA,and MDA levels in the LPRC were markedly lower than the control group.Compared with the control group,the weight-loaded swimming time of mice in the CDDP 0.122 g·kg^-1was significantly prolonged,the liver glycogen content and GSH-PX activity were significantly increased.Chapter 2 Effects of altitude hypoxia exposure in different time on p physical efficiency,mood and cognitive function of miceRandomly divide male BALB/c mice into CON,High altitude hypoxia-3 day group（HAH-3）,and High altitude hypoxia-7 day group（HAH-7）.And each group contained ten mice.Mice in HAH group were place in the multi-factor composite environment simulation medical science experiment cabin,and were decompressed at a constant speed of 5 m/s to the simulated altitude of 6100 m.The hypoxia treatment lasted for 3 days and 7 days.After hypoxia,the physical,emotional and cognitive function of mice were evaluated immediately.1.Effects of high altitude hypoxia on body weight and physical efficiency of miceThe results of the weight change trend of mice in each group showed that compared with the CON group,the weight of the mice in the HIA-3 and HAH-7 groups decreased significantly after the simulated altitude hypoxia exposure for 1 day,and the weight decreased to the lowest after 3 days of hypoxia exposure;After that,the weight of the mice did not change significantly but was still significantly lower than the normoxic control group.The results of autonomous activity showed that compared with the CON group,the autonomous activity distance of HAH-7 mice was significantly increased.It is suggested that simulated altitude hypoxia can cause mice to lose weight and increase autonomous activities without significant effects on physical fitness.2.Effects of high altitude hypoxia on recognition memory in miceNew object recognition test was used to assessment the cognitive ability of mice.The results showed that compared with the CON group,the new object preference index of the HAH-3 group was significantly decreased 1 h after learning,and the new object preference index of the HAH-7 group had no significant change;The new object preference index of mice in the HAH-3 group and HAH-7 group was significantly decreased 24 hours after learning.These results suggest that high altitude hypoxia for 3 days and 7days can cause cognitive impairment in mice,and hypoxia for 3 days is more serious.3.Effects of high altitude hypoxia on anxiety-like behavior in miceThe results of open field experiment and elevated plus maze experiment were used as the evaluation indexes of anxiety-like behaviors in mice.The results of the open field experiment showed that compared with the CON group,the number of entry into the central area of the open field of the mice in the HAH-3 and HAH-7 groups was significantly reduced,and the stay time in the central area was significantly shortened.The results of the elevated plus maze experiment showed that compared with the CON group,HAH-3 mice had a decreasing trend of open-arm entry times,and HAH-7mice had a significant decrease in open-arm entry times;HAH-3 and HAH-7 The open arm stay time of mice was significantly shortened.The above results suggest that the mice of simulated altitude hypoxia for 3 days and 7days can have anxiety-like behaviors,and the anxiety symptoms aggravate with the prolonged exposure time.4.Effects of high altitude hypoxia on depression-like behavior in miceThe results of forced swimming test and tail suspension test as evaluation indexes of depressive behavior in mice.The results of the forced swimming experiment showed that compared with the CON group,the immobility time of the mice in the HAH-3 group and the HAH-7 group was significantly prolonged.The results of the tail suspension experiment showed that compared with the CON group,the immobility time of the HAH-3 group was prolonged,and the immobility time of the HAH-7 group was significantly prolonged.The above results suggest that mice exposed to simulated altitude hypoxia for 3 days and 7 days can have depression-like behaviors,and the depression symptoms aggravate with the prolonged exposure time.The above experiments show that simulated 6100m altitude hypoxia exposure can cause abnormal increase in autonomous activities of mice,anxiety,depression-like behaviors and cognitive impairment.Exposure to6100 m hypoxia for 7 days can be used as a model for drug efficacy evaluation.Chapter 3 Effect of Large plant Rhodiola Capsule on high altitude hypoxia-induced fatigue,mood and cognitive impairmentThe hypobaric hypoxia chamber was used to simulate high altitude hypoxia for 7 days at an altitude of 6100 m.The effects of Large plant Rhodiola Capsule on fatigue,anxiety and depression and cognitive impairment induced by simulated high altitude hypoxia were observed.1 Effects of large plant Rhodiola capsule on body weight and physical efficiency of high altitude hypoxia miceThe results showed that compared with the normoxic control group,on the first day of simulated altitude hypoxia exposure,the weight of the mice in each hypoxia group decreased significantly.With the prolonged hypoxia time,the weight of the mice did not change significantly from the third day but was still significant.Lower than the CON group;the rod time was significantly shortened.The weight-bearing swimming time of mice in the LPRC group and the CDDP group was significantly prolonged;the mice in the LPRC group had a significantly longer stick time than the HAH group.The above results suggest that LPRC can improve the fatigue endurance of high altitude hypoxia mice.2 Effect of Large plant Rhodiola Capsule on high altitude hypoxia induced anxiety-like behavior in miceThe open field test was used to evaluate the anxiety-like behavior of mice.The results showed that compared with the control group,the total distance of open-field movement of mice in the plateau hypoxia group had no obvious change,and the holding time in the central area of open-field was significantly shortened.Each administration group had no significant effect on the total distance of open field movement in plateau hypoxia mice.The residence time in the open field center of mice in the small and large dose LPRC group and CDDP group was significantly prolonged,which was significantly different from that in the HAH group.These results suggest that high altitude hypoxia can induce anxiety-like behavior in mice,and LPRC and CDDP can improve anxiety-like behavior in mice.3 Effect of Large plant Rhodiola Capsule on high altitude hypoxia induced depressive-like behavior in mice.Sucrose preference test was used to evaluate the depressive-like behavior of mice.The Sucrose preference rate of mice in the plateau hypoxia group decreased significantly before and after hypoxia.Compared with the control group,the Sucrose preference rate of mice in the plateau hypoxia group was significantly decreased.The sucrose preference rates of mice in the middle and large doses of LPRC were significantly higher than those in the model group,and the sucrose preference rates of mice in the CDDP,Sal and small doses of LPRC groups showed an increasing trend.The above results suggested that hypoxia at high altitude could lead to depression-like behavior in mice,and LPRC could improve depression-like behavior in mice,while compound CDDP and Sal had no obvious effect.4 Effect of Large plant Rhodiola Capsule on recognition memory in HAH miceNew object recognition test was used to evaluate the memory function of mice.The results showed that compared with the control group,the new object preference indexes of mice in the plateau hypoxia model group were significantly reduced at 1 h and 24 h after learning.Compared with the HAH group,the new object preference index of mice in the CDDP group was significantly increased at 1 h after learning.The new object preference index of the high-dose LPRC was significantly increased at 1 h and 24 h after learning.The new object preference index of new objects at 1 h and 24 h after learning in high-dose LPRC group was also significantly higher than that in Sal group.These results suggest that plateau hypoxia can damage short-term and long-term memory in mice,and long-term memory damage is more serious.LPRC can improve short-term and long-term memory in mice,and CDDP can improve short-term memory in mice.Chapter 4 Mechanism of Large plant Rhodiola Capsule on high altitude hypoxia-induced fatigue,mood and cognitive impairmentIn order to explore the mechanism of Large plant Rhodiola Capsule on improving emotional and cognitive impairment in mice with high altitude hypoxia,the effects of Large plant Rhodiola Capsule on apoptosis of hippocampal neurons,HIF-1α signaling pathway,memory related proteins and synaptic function in mice with hypoxia were observed from histopathological and molecular levels.1 Effect of Large plant Rhodiola Capsule on LTP in Hippocampus PP-DG Region in HAH miceThe changes of LTP in hippocampus of mice were evaluated by in vivo electrophysiological experiment with PS as the index.The results showed that compared with the control group,the hippocampal LTP of mice in the HAH group was significantly damaged.Large dose LPRC group and Sal group significantly improved LTP injury induced by high altitude hypoxia.The above results suggested that Large plant Rhodiola Capsule could improve the decline of synaptic plasticity in hippocampus of mice caused by high altitude hypoxia.2 Effect of Large plant Rhodiola Capsule on the number of Nissl bodies in hippocampus of HAH miceThe damage degree of hippocampal neurons and the number of Nissl bodies were observed by Nissl staining.The results showed that the hippocampal neurons of the control mouse were organized in an organized manner.,with large and round nuclei and obvious nucleoli,and many Nissl bodies were observed.Compared with the control group,the hippocampal area of mice in the HAH group showed increased neuronal shrinkage,deep staining,damaged cell structure,and decreased number of Nissl bodies,especially in the CA1 area.Compared with the HAH group,the number of hippocampal neurons in the LPRC group and the CDDP group recovered to a certain extent,and the change in CA1 area was the most obvious.The above results suggested that acute high altitude hypoxia exposure could lead to pathological damage of brain tissue,and LPRC and CDDP could protect brain tissue damage caused by hypoxia.3 Effect of Large plant Rhodiola Capsule on the expression of BDNF-Trk B Signaling Pathway and Synaptic Related Proteins in Hippocampus of HAH miceChanges in hippocampal protein of BDNF-Trk B signaling pathway levels were detected by Western blotting.Western blot was used to detect the expression of BDNF-Trk B signaling pathway and synaptic related proteins in the hippocampus of high altitude hypoxia mice.The results showed that compared with the control group,the level of Trk B protein in the hippocampus of mice in the HAH group was significantly increased,and the levels of BDNF and PSD-95 proteins were decreased.CDDP and middle and large doses of LPRC can significantly increase the protein level of BDNF in hippocampus of mice.The expression of Trk B,PI3K,AKT and PSD-95 in LPRC had an increasing trend,CDDP had no significant effect,Sal had an increasing trend.These results suggest that high altitude hypoxia can lead to the loss of memory-related proteins and the impairment of synaptic function in mice.The protective effect of LPRC on hypoxic emotion and cognitive impairment may be mediated by the regulation of memory-related proteins and synaptic structural proteins.4 Effect of Large plant Rhodiola Capsule on the expression of HIF-1α?related proteins in hippocampus of HAH miceChanges in hippocampal protein levels were detected by Western blotting.Compared with the control group,The results showed that the HIF-1α?protein level in the hippocampus of mice in the HAH group had no significant change,the PHD-2 protein level was significantly decreased,the VEGF protein level was increased,and the FIH and EPO protein levels were decreased,but there was no significant difference.Compared with the HAH group,the HIF-1α?and EPO protein levels in the hippocampus of mice in the LPRC showed an increasing trend,while the FIH and VEGF protein levels showed a decreasing trend,and there was no significant effect on the PHD-2 protein level.These results suggest that LPRC may inhibit FIH to rapidly up-regulate HIF-1α?protein level,restore VEGF level and promote EPO expression,thereby improving the body’s hypoxia tolerance.Through the above research,this study mainly got the following conclusions:（1）The fatigue,anxiety,depression-like behavior and cognitive impairment of mice could be induced by hypoxia exposure for 3 and 7 days at 6100 m altitude in hypobaric chamber,and the effect of high altitude environment on neurological function was related to the time of hypoxia exposure.（2）Large plant Rhodiola Capsule could effectively improve the fatigue,anxiety,depression and cognitive impairment of mice induced by high altitude hypoxia.It is suggested that Large plant Rhodiola Capsule may be a preventive and therapeutic drug for fatigue,emotional and cognitive dysfunction caused by plateau hypoxia.（3）Large plant Rhodiola Capsule can improve the synaptic plasticity of hippocampal neurons and protect brain neurons in high altitude hypoxia mice,and the mechanism may be related to the regulation of HIF-1α signaling pathway,BDNF-Trk B and PSD-95 protein expression.