| Purpose To observe the therapeutic effect of Bao Shen Recipe(BSF)on spontaneous DN mice and explore the mechanism of intestinal bacteria regulationMethods One hundred and twenty-five 6-week-old db/db diabetic mice were randomly divided into 5 groups:db/db-m(DN model group);db/db-hd(BSF high-dose treatment group);db/db-md(BSF medium-dose treatment group);db/db-ld(BSF low-dose treatment group),db/db-me(positive control group);DN model was constructed,db/m of wild mice was used as control(blank control group);db/db-me was given irbesartan as positive control;at the end of 8 weeks of age,high,medium,and low-dose BSF treatments were given,respectively,and the positive control group was given irbesartan treatment for 26 weeks,during which body weight was recorded every 4 weeks,random blood glucose,urinary microalbumin excretion rate,blood urea nitrogen,and serum creatinine were measured,and the animals were sacrificed at 18 weeks of treatment,and renal tissues were obtained for HE,Masson,and PAS staining to observe renal pathological changes,and renal tissue homogenates were prepared from the other kidney.The levels of IL-6,TNF a and TNF-1R and TNF-2R in the kidneys of different groups of mice were measured by ELISA.At the same time,fecal samples were taken at early,middle,and late stages of treatment(1,8,and 18 weeks of administration)and high-throughput sequencing was performed using 16S rRNA to observe changes in intestinal bacterial structural diversityResults①db/db mice were fed for 8 days,the urinary albumin excretion rate was significantly increased,and the DN model was successfully constructed.② At 1,4,8,12 and 18 weeks of db/db-m administration,the body weight level and blood glucose of the mice were significantly increased compared with those of db/m mice in the same week group,with statistical differences(P<0.01);at 4 weeks of administration,the body weight level and blood glucose excretion of the db/db-hd group were decreased compared with those of the db/db-m group,with statistical differences(P<0.01).Urinary microalbumin and serum creatinine were significantly increased in the other groups at 20 weeks of age(P<0.01).The levels of blood urea nitrogen and serum creatinine in db/db-me group were decreased(P<0.01),and the levels of blood urea nitrogen and serum creatinine in high and medium dose BSF groups were significantly lower than those in db/db-me group(P<0.01)①Renal pathology after 18 weeks of drug treatment showed diffuse proliferation of glomerular cells and stroma,basement membrane thickening,granular and vacuolar degeneration of tubular epithelium,and scattered inflammatory cell infiltration in the db/db-m group,and high and medium doses of BSF were able to reduce the matrix expansion score(P<0.05),and the degree of reduction was greater than that in the db/db-me group,with a statistically significant difference(P<0.05)④,after 4 weeks of drug treatment,IL-6,TNFα,TNF-1R,and TNF-2R levels decreased in the BSF low-dose,medium-dose,and high-dose treatment groups compared with the db/db-m group(P<0.05),with a more significant decrease in the db/db-hd group(P<0.01)⑤ ter analysis of the number of species categories,the number of species showed a significant increase after 16 weeks of feeding in db/db-m group mice compared with db/m group mice,with a statistically significant difference(P<0.05)After 8 weeks of drug treatment,the diversity and abundance of bacteria in db/db-hd mice were significantly lower than those in db/db-me group(P<0.05),especially high-dose BSF could make the number of intestinal bacteria species in DN mice close to that in normal control mice.After 16 weeks of drug treatment,there was no difference in the number of intestinal bacteria species in the other groups of db/db-m(P>0.05).Compared with db/m mice,there was no difference in the number of intestinal bacteria species in the high-dose BSF(P>0.05),and other groups showed increased intestinal bacteria(P<0.05)⑥ The specific differential bacteria were analyzed,and 1 week after early dru g treatment,Verrucomicrobia、Akkermansia、Lachnospiraceae_NK4A136_group were significantly higher in the db/db-m group than in the normal control grou p(P<0.05).Compared with db/db-m group,the above flora was decreased in different doses of BSF(P<0.05).while the abundance of Ruminococcaceae UC G 014in the Ruminococcaceae group was increased(P<0.05).In the middle stage of 8-week treatment with drugs,the abundance of Alistipes and Lachnosp iraceae NK4A136_group were reduced in the db/db-hd group compared with th e db/db-m group(P<0.05).At the late stage of 18 weeks of drug treatment,Alistipes and Lachnospiraceae_NK4A136_group increased in db/m(P<0.05),a nd unidentified Enterobacteriaceae increased in db/db-me(P<0.05)Conclusion BSF can delay the progression of diabetic nephropathy,effectively reduce the excretion rate of blood glucose,creatinine and urinary microalbumin,and improve renal pathology in DN model,and the mechanism may be related to the regulation of intestinal flora and the inhibition of a series of inflammatory factors.This study provides some theoretical basis for the clinical translation of BSF. |
PDF Full Text Request