| Objective:To investigate the establishment method of diarrhea mice with kidney-yang deficiency syndrome,to establish a mouse model of diarrhea with kidney-yang deficiency syndrome,to study the gut microecological mechanism of Sishen pills decoction in the process of kidney-gut impairment in diarrhea with kidney-yang deficiency syndrome.Methods:SPF-grade Kunming male mice were used as the research objects.（1）Different doses of adenine suspension were used for gavage,and the mice were tested for energy metabolism,sexual function,organ index,kidney structure and function,colonic structure,oxidative stress and gut content microbiota.（2）Adenine suspension combined with Folium sennae decoction for different doses and days was used to detect the indexes of mouse kidney structure and function,gut content enzyme activity and microbial activity.（3）Adenine suspension combined with Folium sennae decoction was used to construct the diarrhea mice with kidney-yang deficiency syndrome,and Sishen pills decoction was used to intervene.The indexes of mouse organ index,kidney structure,energy metabolism were detected.（4）After the successful modeling of diarrhea mice with kidney-yang deficiency syndrome in method（3）,mice were tested for kidney function,intestinal inflammatory response,gut content microbiota and SCFAs.（5）After the intervention of Sishen pills decoction in method（3）,the mice were tested for kidney function,intestinal inflammatory response,gut content microbiota and SCFAs.Results:（1）Compared with NC group,serum c AMP level in NMM group was decreased（P<0.05）,cGMP level was increased（P<0.05）,cAMP/c GMP was decreased（P<0.05）,T and E2levels were decreased（P<0.05;P<0.05）,serum Cr and BUN were increased（P<0.01;P<0.01）,MDA level in kidney was increased（P<0.01）,SOD level was decreased（P<0.01）and the structural damage of kidney and colon tissues were obvious.The enrichment of different characteristic bacteria appeared in mice after different doses of adenine.（2）Compared with C group,body weight of mice in M2 group decreased（P<0.05）,serum Cr level increased（P<0.05）,BUN level decreased（P<0.05）,lactase decreased（P<0.01）,sucrase and microbial activity increased（P<0.01;P<0.01）.（3）Compared with CC group,the body weight and anal temperature of mice in CM group decreased（P<0.01;P<0.01）,diarrhea index increased（P<0.01）.Serum cAMP level was decreased（P<0.01）,cGMP level was increased（P<0.01）,cAMP/cGMP was decreased（P<0.01）,and kidney structure damage was obvious.Compared with CX group,body weight and anal temperature in CS group were changed（P>0.05;P>0.05）,diarrhea index decreased（P<0.01）.Serum c AMP,cGMP and c AMP/c GMP were changed（P>0.05;P>0.05;P>0.05）,the damage of kidney structure was alleviated.（4）The characteristic bacteria Lactobacillus intestinalis and Bacteroides acidifaciens in CM group were enriched.After modeling,SCFAs in gut content of mice was inhibited（P<0.01;P<0.01;P<0.01;P<0.01;P<0.01;P<0.01）,and the levels of serum Cr,BUN,IL-6,TNF-αand s Ig A in colon tissues were changed.Lactobacillus intestinalis and Bacteroides acidifaciens were negatively correlated with acetic acid,positively correlated with Cr,and negatively correlated with IL-6 and BUN.（5）The characteristic bacterium Lactobacillus johnsonii in CS group was enriched,and SCFAs were changed.The serum Cr was decreased（P>0.05）,BUN was increased（P>0.05）,the levels of IL-6and TNF-αwere decreased（P<0.01;P<0.05）,s Ig A in colon tissue increased（P<0.01）.Lactobacillus johnsonii and SCFAs were correlated with IL-6,TNF-αand s Ig A.Conclusion:（1）The kidney-yang deficiency syndrome mouse model was successfully established by gavage of 50 mg/（kg·d）adenine for 14 d.（2）Gavage of 50 mg/（kg·d）adenine for 14 d combined with 10 g/（kg·d）Folium sennae for 7 d could significantly damage kidney and gut functions in mice.（3）Diarrhea mice model with kidney-yang deficiency syndrome was successfully established by gavage of 50 mg/（kg·d）adenine for 14 d combined with 10 g/（kg·d）Folium sennae for 7 d.（4）Diarrhea with kidney-yang deficiency syndrome altered the structure and function of gut content microbiota in mice,inhibited SCFAs,damaged kidney structure and function and activated intestinal inflammatory response.The interaction between Lactobacillus intestinalis and Bacteroides acidifaciens with SCFAs,intestinal inflammatory response and kidney function might be the mechanism of gut-kidney impairment during diarrhea with kidney-yang deficiency syndrome.（5）Sishen pills decoction changed the structure and function of gut microbiota,promoted propionic acid,butyric acid,isobutyric acid and isovaleric acid,inhibited intestinal inflammatory response and improved the damage of kidney structure and function.The interaction between the characteristic bacterium Lactobacillus johnsonii and propionic acid,intestinal inflammatory response,and kidney function may be the mechanism of action of Sishen pills in the treatment of diarrhea with kidney-yang deficiency syndrome. |
