Experimental Research On The Correlation Between "antibiotic-related Diarrhea" And "spleen-yang Deficiency Syndrome" Based On Metabonomics

Antibiotic-associated diarrhea(AAD)belongs to the category of "diarrhea" in TCM.Diarrhea is based on spleen deficiency.The spleen is the acquired basis,the source of Qi and blood biochemistry,and the pivot of Qi elevation and descent.The spleen has the functions of transporting,clearing and collecting blood.Spleen-yang is the active part of Yang in spleen-qi.By its warm,dynamic,upward and outward functions,it can maintain the function of the spleen to transport,nourish and nourish,and promote the normal operation of the spleen.The deficiency of Spleen-yang leads to the weakness of the valley of transport and transformation,the weakness of the essence of transport and transformation,which turns into wetness and turbidity,the clearing of Yang does not rise,and the wetness sinks and diarrhea occurs.Long-term unreasonable use of antibiotics leads to accumulation of toxic drugs,depletion of spleen and stomach qi,vulnerability of spleen yang,inappropriate spleen movement,no valley,adverse Qi machine,imbalance of ascent and descent,no ascent of Qingqi,no descent of turbidity,endogenous turbidity,which leads to antibiotic-related diarrhea.Many modern physicians believe that "spleen-yang deficiency" is closely related to AAD,and the legislation of "warming the middle and strengthening the spleen" has achieved good results.However,there is a lack of data to support the internal relationship between them.Therefore,based on metabonomics,this study constructed the AAD model induced by antibiotics and the Spleen-Yang deficiency model induced by bitter cold drugs.The serum of SD rats was detected by ultra-high performance liquid chromatography-four-stage time-of-flight mass spectrometry(UPLC-Q-TOF).Endogenous markers of different models were screened out,and the relationship between AAD and Spleen-Yang deficiency syndrome was explored from the metabolic level.Basis and data support.PurposeTo investigate the relationship between antibiotic-related diarrhea and Spleen-Yang deficiency syndrome in two groups of rats by observing the general situation and stool changes of antibiotic-related diarrhea and Spleen-Yang deficiency syndrome,and to screen endogenous markers of two groups of animal models by using metabonomics,and to explore the correlation between AAD and Spleen-Yang deficiency syndrome from the metabolic level.And provide the relevant basis for internal links.MethodsThirty SD rats were randomly divided into three groups:blank control group,antibiotic-related diarrhea group and Spleen-Yang deficiency group,with 10 rats in each group.The models were made with saline,clindamycin and rhubarb respectively.Abdominal aortic blood of SD rats was collected after successful modeling,and the serum of SD rats was detected by ultra-high performance liquid chromatography-four-stage time-of-flight mass spectrometry(UPLC-Q-TOF).Data were analyzed by SPSS20.0 software,and metabolomics data were analyzed and counted by markerlynx software to confirm endogenous differentials.Metabo Analysis 3.0was used to identify the metabolic pathways of potential biomarkers.Results1.Weight changes:On the 10th day of modeling,the weight of AAD group was significantly higher than that of blank control group(p=0.021);the weight of Spleen-Yang deficiency group was significantly higher than that of blank control group(p=0.002);the weight of AAD group was not significantly different from that of Spleen-Yang deficiency group(p=0.229).2.2 hours of total fecal weight:On the 10th day of modeling,compared with the total fecal weight of rats in the blank control group,the total fecal weight of rats in the AAD group was p=0.031 with statistical significance;compared with the blank control group and the AAD group,the total fecal weight of rats in the Spleen-Yang deficiency group had no statistical difference(p>0.05).3.Metabonomics analysis results:22 biomarkers were identified in AAD group compared with blank control group,and 21 biomarkers were identified in Piyang deficiency group compared with blank control group.Seven endogenous biomarkers were screened out from AAD group and Spleen-Yang deficiency group:homoserine O-phosphate,glycine glutarate,dodecyl carnitine,VPGPR enterostatin,DG(14:0/14:0/0:0),D Bilin and PI(16:0/18:2(9Z,12Z)4.Metabolic pathways:The pathways with higher influencing factors in AAD-related metabolic pathways are Valine,leucine and isoleucine degradation(degradation of valine,leucine and isoleucine);the pathways with higher influencing factors in Spleen-Yang deficiency-related metabolic pathways are Pentose and glucuronate interconversions(mutual conversion of pentose and glucuronate).Conclusions1.Apparent characteristics and TCM syndromes,such as general condition,weight change and stool condition after successful modeling,show that AAD is correlated with Spleen-Yang deficiency syndrome(rhubarb modeling).2.The results of serological analysis of metabonomics showed that AAD and Spleen-Yang deficiency syndrome(rhubarb model)had some same endogenous markers,and there was a certain correlation at the level of metabonomics.3.Metabonomic analysis showed that AAD was not identical with endogenous markers of Spleen-Yang deficiency syndrome,but related to the difference of metabolic pathways of AAD and Spleen-Yang deficiency syndrome.4.The results of metabolic pathway suggest that AAD is related to amino acid metabolism(rhubarb model)and Spleen-Yang deficiency is related to glucose metabolism.