Study On The Antidepressant And Anti-fatigue Effects Of The Total Flaconoids Extracted From Xiaobuxin-Tang(XBXT-2) And Its Mechanism

Objective:Gut microbiota is closely related to the pathogenesis of depression.Previous studies have found that XBXT-2 have antidepressant effects,and this study aimed to explore whether the antidepressant effect of XBXT-2 are related to the regulation of gut microbiota.Fatigue is the core physical symptom of depression,and depression and fatigue have a high prevalence.On this basis,we explored whether XBXT-2 has anti-fatigue effect and its mechanism.Methods:1.Evaluation of the antidepressant effect of XBXT-2:Wistar rats were divided into control,model,FXT(10 mg/kg)and XBXT-2(100 mg/kg)groups to prepare the CUS model.The SPT,NSFT and OFT were used to evaluate the antidepressant activity of XBXT-2.2.Evaluate the antidepressant mechanism of XBXT-2 on the CUS model:The composition of gut microbiota of rats was analyzed by 16s RNA pyrosequencing.WB and ELISA were used to detect the levels of immunocellular marker IBA-1,inflammatory mediator TLR-4 and various inflammatory factors in the hippocampus and colon of rats.3.Evaluation of anti-fatigue effect of XBXT-2:C57BL/6J mice were divided into control,caffeine(10 mg/kg)and high,middle and low dose XBXT-2 groups(200、100、50 mg/kg)to establish exercise fatigue model.The anti-fatigue effect of XBXT-2 was evaluated according to behavioral indexes such as the exercise distance and exercise time in the running platform experiment.C57BL/6J mice were divided into control,LPS(0.2 mg/kg),LPS+caffeine(10mg/kg)and high,middle and low dose LPS+XBXT-2 groups(200、100、50mg/kg)to make LPS fatigue model.The anti-fatigue effect of XBXT-2 was evaluated according to the behavioral indexes such as the exercise distance and exercise time in the running platform experiment.4.The anti-fatigue mechanism of XBXT-2 was evaluated on LPS fatigue model and antibiotic interference model:C57BL/6J mice were divided into control,LPS(0.2 mg/kg),LPS+caffeine(10 mg/kg)and LPS+XBXT-2 groups(100 mg/kg)to make LPS fatigue model.The levels of monoamine transmitters in different brain regions were determined by HPLC.The levels of NH3,GLU,BUN,LAC,LDH and CK in serum of mice were detected by blood biochemical analyzer.Glycogen kit was used to detect glycogen content in liver and muscle.The levels of IL-4,IL-6,IL-1β and TNF-α in muscle tissues of mice were detected by ELISA.The gut microbiota of mice was analyzed by 16s RNA pyrosequencing.C57BL/6J mice were divided into control,caffeine(10 mg/kg),XBXT-2(100 mg/kg),ABX,ABX+caffeine(10 mg/kg),and ABX+XBXT-2 groups(100 mg/kg)to make antibiotic interference model in mice.The anti-fatigue effect of XBXT-2 was evaluated according to the behavioral indexes such as the exercise distance and exercise time in the running platform experiment.Results:1.Evaluation of the antidepressant effect of XBXT-2:Behavioral test results showed that XBXT-2(100 mg/kg)could significantly increase sucrose preference and number of standings in OFT,and significantly reduce the latency to enter the central in OFT and the latency to eat in NSFT.2.Evaluate the antidepressant mechanism of XBXT-2 on the CUS model:Sequencing analysis of gut microbiota showed that the relative abundance ratio of Firmicutes to Bacteroidetes decreased in XBXT-2 groups,while the number of beneficial bacteria increased and the number of pathogenic bacteria decreased.WB and ELISA results showed that the levels of IBA-1,TLR-4,IL-1β and IL-6 in hippocampus and IBA-1,TLR-4,IL-1β,IL-6 and TNF-α in colon of rats after XBXT-2 treatment were significantly decreased,while the level of IL-10 was significantly increased.3.Evaluation of the anti-fatigue effect of XBXT-2:The results of exercise-induced fatigue model showed that single administration of XBXT-2(100 mg/kg)and caffeine(10 mg/kg)could significantly increase the exercise distance and exercise time of normal mice in the running platform experiment.The results of LPS fatigue model showed that single administration of XBXT-2(50,100 mg/kg)and caffeine(10 mg/kg)could significantly increase the exercise distance and time of LPS fatigue model mice.4.Evaluate the anti-fatigue mechanism of XBXT-2 on the LPS fatigue model and antibiotic interference model:The results of LPS fatigue model showed that single administration of XBXT-2(100 mg/kg)can significantly increase the 5-HT,DA in the hypothalamus,NH3,GLU in serum,liver glycogen in the liver,muscle glycogen,IL-6,IL-1β,TNF-α in muscle and Campilobacterota,Bifidobacterium,Escherichia-Shigella bacteria and the relative abundance of the Firmicutes/Bacteroidetes ratio in gut;The bacteria of Deferribacterota,Campylobacter and Mucispirillum in gut of LPS fatigue model mice were significantly reduced.The results of antibiotic interference model showed that single administration of XBXT-2(100 mg/kg)and caffeine(10 mg/kg)significantly increased the distance and time of exercise in fatigue model mice,ABX+caffeine(10 mg/kg)and ABX+XBXT-2(100 mg/kg)significantly increased the distance and time of exercise in mice with antibiotic interference.XBXT-2(100 mg/kg)and ABX+XBXT-2(100 mg/kg)had no effect on spontaneous activity trajectory of fatigue model mice.Conclusion:1.Based on the rat CUS model,XBXT-2 has anti-depression and anti-anxiety effects,which are related to the regulation of gut microbiota.2.Based on the LPS fatigue model of mice,XBXT-2 has anti-fatigue effect,which is related to the regulation of energy metabolism,inflammatory factors,neurotransmitters and gut microbiota.Figure[21]table[6]reference[53]...