| Objective:With the fast pace of modern society and increase of life pressure,fatigue has become a state not being neglected.As a pathogenic factor,fatigue might lead to a variety of diseases.Researches displayed that the incidence of the spleen qi deficiency pattern was high in people with fatigue.Based on analysis of fatigue phenomena in tired people,causes of the spleen deficiency triggered by tired people and by traditional fatigue mode were different.By researching on the disease and mechanism of learning and memory ability decreasing due to the spleen deficiency caused by fatigue was of great importance in explorations of intervening measures on symptoms differentiated as the spleen deficiency and pathological characteristics in tired people.The pathogenesis in traditional Chinese medicine(TCM)of the spleen deficiency caused by fatigue was originated from the theory of exhaustion-fatigue damaging the spleen.The pathogenesis of the spleen deficiency due to fatigue in TCM influencing on learning and memory ability was complied with the theory of the spleen storing reflection.The biological mechanism was ascribed to imbalance of central nervous system pathways induced by GABA and GLu.Therefore,under the support of State Administration of Traditional Chinese Medicine of the People’s Republic of China,Traditional Chinese Medicine on Navigation a key discipline of Beijing University of Chinese Medicine(BUCM)and Collaborative Innovation Center of BUCM on TCM navigation,by researching on ancient literatures and modern documents aimed at study of the cause and pathogenesis of the spleen deficiency and explanations of the relationships between fatigue and spleen deficiency,and spleen deficiency and learning and memory ability.Based on this,the TCM theory with guiding significance and related mechanism of neurophysiology were summarized targeting on spleen fortifying formula Buzhong Yiqi decoction and liver soothing and spleen fortifying formula modified Tifukang in the treatment of the spleen deficiency due to fatigue,further to tidy up and perfect the TCM theories of exhaustion-fatigue damaging the spleen,the spleen storing reflection,and the spleen corresponding with thought based on the five minds,and finally establish the theoretical research framework.Meanwhile,based on the theory of exhaustion-fatigue damaging the spleen and clinical characters of the spleen deficiency caused by fatigue in modern population,rat model of the spleen deficiency due to fatigue by modified multiple plate-form method(MMPM)was established.From the perspectives of the spleen storing reflection and the spleen corresponding with thought based on the five minds,Buzhong Yiqi decoction a classical formula on treating the spleen deficiency was selected as a targeting Chinese medicine group,and modified Tifukang a liver-soothing and spleen-fortifying prescription was chosen as a TCM control group.By integral experiment,targets of pharmacological effects by Buzhong Yiqi decoction and modified Tifukang in the treatment of decrease of learning and memory ability in rats in spleen deficiency due to fatigue were clearly stated in order to explore neurobiological mechanisms in different levels and lay the research foundation for thought of diagnosis and treatment in developing combination of disease and symptom.Methods:1.By retrieving and research on relevant literatures from the ancient works,China National Knowledge Internet(CNKI in Web version)ranging from 1979 to 2016,and VIP(Web version)ranging from 1989 to 2016 with the keywords of exhaustion-fatigue damaging the spleen,spleen deficiency,spleen storing reflection,and the spleen corresponding with thought based on the five minds,the understanding of exhaustion-fatigue damaging the spleen,spleen storing reflection,and the spleen corresponding with thought based on the five minds in TCM theory were combed by statistical analysis on relevant literatures and the correlation among the fatigue,spleen deficiency,and learning and memory ability was further studied.2.Exploring and establishing the ideal animal model of exhaustion-fatigue damaging the spleen were used for intervening measures of the spleen deficiency pattern,causes and pathological characteristics of the spleen deficiency,and further for modern mechanism study of promoting on the spleen deficiency syndrome.3.Dynamic observations on rat model with the spleen deficiency caused by fatigue and regulating effects of Buzhong Yiqi decoction and modified Tifukang on symptoms of rat’s spleen deficiency and learning and memory ability from different levels of behavioristics,histomorphology,and molecular biochemistry explore its biological mechanisms.Results:1.Experimental test from rat model of the spleen deficiency due to fatigue displayed that the ideal experimental model of the spleen deficiency due to fatigue was the small platform with water beneath group of 12 hours per day for 21 days,whose symptomatic scores of the spleen deficiency pattern were in accordance with the standard of spleen qi deficiency pattern model.2.By Morris water maze,the escape latency in the small platform with water beneath group of 12 hours per day for 21 days was much longer than that in the blank control group.Times of crossing the platform and percentage of distance in target quadrant were greatly decreased in comparison with those in the blank control group with statistical differences.Compared with model group,the latency was shortened and times of crossing the platform and percentage of distance in target quadrant were increased in Buzhong Yiqi decoction group and low-dosage of Tifukang group,which were displayed statistical differences.It was displayed that spleen-fortifying formula of Buzhong Yiqi decoction and low-dosage of Tifukang of liver-soothing and spleen-fortifying prescription both had improvement effect on rat’s learning and memory ability in the level of behavioristics.3.Histomorphological test results displayed GABA and GLu receptors in hippocampal CA1 area and striatal area in rats from different groups that content of GABAal,GABAAa5,NR1,and NR2A receptors in hippocampal CA1 area in spleen deficiency due to fatigue model group(abbreviation as model group)was greatly increased compared with those in the blank control group.In striatal area,in comparison with 21-day blank control group,content of GABAα1,GABAAα5,NR1,and NR2A receptors in rats in model group was significantly increased.Compared with model group,low-dosage of Tifukang could decrease the content of GABAal and GABAAa5 receptors in hippocampal CA1 area and striatal area and increase the content of NR1 and NR2A receptors in hippocampal CA1 area and striatal area in rats.4.The detection results of gene expressions in GABA and GLu receptors in hippocampal CA1 area and striatal area in rats in each group displayed that mRNA expressions in GABAα1 and GABAAa5 receptors in the hippocampal CA1 area in the model group were obviously increased compared with those in the blank control group,while mRNA expressions in NR1 and NR2A were decreased compared with those in the blank control group.In the striatal area,in comparison with 21-day blank control group,mRNA expressions in GABAal and GABAAa5 receptors in model group were significantly increased in comparison with those in the blank control group,while mRNA expressions of NR1 and NR2A were lower compared with those in the blank control group.In comparison with the model group,mRNA expressions in NR1 and NR2A receptors in hippocampal CA1 area and striatal area were greatly increased in the medium dosage of Tifukang group.In the low-dosage of Tifukang group,mRNA expressions in GABAal and GABAAα5 receptors in hippocampal CA1 area and striatal area were greatly decreased,while in NR1 and NR2A receptors were significantly increased with a better effect.5.The detection results of protein expressions in GABA and GLu receptors in hippocampal CA1 area and striatal area in rats in each group displayed that receptor protein expressions of GABAal,GABAAa5,NR1,and NR2A in the hippocampal CA1 area in the model group were obviously increased compared with those in the blank control group.In the striatal area,in comparison with 21-day blank control group,content of GABAα1,GABAAα5,NR1,and NR2A receptors was increased in rats in the model group.Compared with the model group,Buzhong Yiqi decoction could decrease GABAα1 and GABAAa5 protein expressions in hippocampal CA1 area and striatal area and up-regulate NR1 and NR2A receptor protein expressions with a better effect.In the low-dosage of Tifukang group,effect of decreasing GABAal and GABAAa5 protein expression and increasing NR1 and NR2A protein expression were also great.Conclusion:1.By small platform with water beneath,spleen-deficiency pattern model due to fatigue can be established and small platform with water beneath group of 12 hours per day for 21 days is the most ideal model.2.Spleen-fortifying formula of Buzhong Yiqi decoction can obviously improve spleen-deficiency symptoms in the model group whose effect is superior to that by modified Tifukang with actions of soothing the liver and fortifying the spleen,which indicates that with regard to spleen-deficiency symptoms in rats in the model group,method of spleen-fortifying is superior to that of soothing the liver and fortifying the spleen.3.Learning and memory ability is decreased in rats in the model group.Both Buzhong Yiqi decoction and low-dosage of Tifukang can both greatly improve decrease of learning and memory ability in rats in spleen deficiency due to fatigue.Effect by low dosage of Tifukang is better than that by Buzhong Yiqi decoction,which suggests that methods of fortifying the spleen and soothing the liver and fortifying the spleen both improve learning and memory ability to a certain degree and effect of soothing the liver and fortifying the spleen is more ideal.4.In the model group,variation tendencies of receptor content,gene level and protein level in GABAal and GABAAa5 receptors in hippocampal CA1 area and striatal area in rats are tantamount with a remarkable increase.mRNA expressions in NR1 and NR2A receptors are both decreased,but the content of receptors and protein expressions were increased.It is indicated that increase of GABA due to long time model intervention leads to fatigue in rat’s body and nerve center.In the development of fatigue,increase of GABA and its receptor levels can indirectly causes increase of excitatory neurotransmitter and its receptor level.Inhibition process takes the superiority by inhibitory neurotransmitter like GABA in central nervous system,which leads to the occurrence of fatigue in nerve center.5.Spleen-fortifying formula of Buzhong Yiqi decoction and low-dosage of Tifukang a liver-soothing and spleen-fortifying prescription both can improve poor learning and memory ability due to spleen deficiency caused by fatigue,which indicates that rats in spleen deficiency due to fatigue presents with liver depression to some extent by drugs counterevidence.TCM intervention by fortifying the spleen and soothing the liver and fortifying the spleen both can improve the learning and memory ability in model rats.The action point may relate with regulatory pathways of GABA and GLu neurotransmitter receptors,and it is speculated that regulating effect on inhibitory neurotransmitter of GABA and its receptors is more remarkable. |
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