Protective Mechanism Of Yishenhuashi Granule On Diabetic Kidney Disease Based On The Theory Of ‘Gut-Kidney Axis’

Objective: By collecting the DKD patient’s general condition and four diagnostic data,were retrospectively analyzed in patients with TCM syndrome distribution.On this basis,the intervention effect of Yishenhuashi Granules on intestinal microflora of diabetic nephropathy model rats was observed,and to explore the therapeutic effect and mechanism of Yishenhuashi granule on DKD based on the “gut-kidney axis” theory.Methods:1.Clinical studies using retrospective study method,collection and screening of DKD patients’ basic situation,the Chinese and western medicine diagnosis,symptoms and four diagnostic information and so on,carried out on the patients’ basic information,syndrome type frequency statistical analysis to study the TCM syndrome features of diabetic nephropathy,the distribution situation,provide a theoretical basis for study of animal experiment next;2.DKD rat model was constructed by high-sugar and high-fat diet combined with low-dose strepzotocin,and randomly divided into normal group(N),model group(M),Yishenhuashi Granule low-dose group(YL),high-dose group(YH)and valsartan group(V).YL,YH and V groups were given 2.77g/(kg·d),5.54g/(kg·d)and 7.38mg/(kg·d)intragastric administration,respectively,and N and M groups were given the same amount of normal saline.The general condition,body weight and blood glucose of the rats were observed.Six weeks later,blood was collected under anesthesia to detect renal function,blood lipid and inflammatory indexes,and the pathological changes of colon and kidney were observed.Total microbial DNA of feces was extracted and sequenced in 16 S r DNA V3+V4 region.Results: 1.Clinical study:(1)A total of 745 DKD patients were included in this study,including 479 male patients and 266 female patients,with a male to female ratio of 1.80:1.(2)The mean age of the patients was(63.65±12.87)years,and 34 patients were under 40 years old,accounting for 4.56% of the total number of patients;291 patients aged 40-59 years,accounting for 34.77%;There were 434 patients aged 60-79,accounting for 51.14%of the total number of patients.There were 83 patients over 80 years old,the total number of patients was 9.53%.(3)The distribution of patients at different stages was analyzed,including 179 patients with DKD stage Ⅰ,accounting for 24.03% of the total number of patients;52 patients with DKD stage Ⅱ,accounting for 6.98%;188 patients with DKD stageⅢ,accounting for 25.23% of the total;120 patients with DKD stage Ⅳ,accounting for16.11% of the total number of patients;There were 206 DKD stage Ⅴ patients,accounting for 27.65% of the total.(4)Frequency analysis of clinical symptoms showed that fatigue,poor sleep,blurred vision,foamy urine,dry mouth and lower limb edema(frequency >30%)were the most common clinical symptoms of DKD.The tongue is mainly red,dark red or dark,light red or light,and the coating on the tongue is thin and white,yellow and greasy,yellow or thin,white and greasy or white,and the pulse is mainly stringy and deep.(5)Syndrome distribution,the deficiency of spleen and kidney was the main syndrome,a total of 317 cases,accounting for 42.55%;There were 181 cases of deficiency of lung and kidney qi(24.30%).122 cases of qi and Yin deficiency syndrome,accounting for16.38%;78 cases of liver and kidney deficiency syndrome(10.47%);47 cases,accounting for 6.31%.A total of 530 patients(71.14%)were combined with standard demonstration,including 197 cases(26.44%)of blood stasis syndrome.There were 151 cases(20.27%)of phlegm and turbidity syndrome.There were 97 cases of damp-heat syndrome,accounting for 13.02%.2.Animal experiments:(1)Body weight,blood glucose and other general conditions:the body weight of rats in the medication group increased slowly with time,and the body weight of YH group was significantly higher than that of YL group(P<0.01).After 6 weeks of treatment,blood glucose in YH group was significantly lower than that in M group(P<0.01),but there was no significant difference compared with YL group(P<0.05).(2)UACR,renal function,blood lipid and inflammation indexes: Compared with M group,UACR results in drug group were significantly decreased(P<0.01),and YH group was lower than YL group(P<0.05).Serum BUN,Cys C,UNAG,TG,TC,LDL,HDL and IL-6of rats in M group were significantly higher than those in N group(P<0.01),T-SOD significantly decreased;Compared with the M group,the indexes of the medication group were improved in different degrees(P<0.01 or P<0.05),compared with YL group,the levels of BUN,TC and TG in YH group were significantly decreased,while the levels of Cys C,LDL and IL-6 were extremely significantly decreased.In addition,there was no significant difference in Scr between the 5 groups.(3)Pathological results showed that there was no significant morphological difference in colonic HE staining of rats in each group.Proliferation of renal mesangial cells and matrix and vacuolar degeneration of tubules were reduced in YH group compared with M group,and ZO-1 expression was increased in colonic tissue.(4)Microbial sequencing: The diversity and abundance of bacteria in group M were lower than those in group N,and the diversity and abundance of bacteria in group YH were higher than those in group M.PCo A analysis showed that the aggregation degree of the M group was poor and outlier samples appeared.Compared with the M group,the aggregation degree of the YH group was significantly recovered,and the distance between the YH group and the N group was closer.The dominant bacteria in each group were Bacteroidetes and firmicutes.The F/B value of rats in M group was significantly up-regulated compared with that in N group,and the F/B value of rats in YH group was decreased compared with that in M group.(5)Species differences: LEf Se analysis and comparison showed that the dominant species in group M were Anaerospira and Prevococcus UCG＿001,the dominant species in group YH were Lactobacillus,Ruminococcus,Clostridium and Subdoligranulum,and the dominant species in group YH were Lactobacillus muri and Ruminococcus sp＿YE281.(6)Functional prediction: At KEGG L2 level,the abundance of functional genes of carbohydrate metabolism,membrane transport,signal transduction,lipid metabolism,DNA transcription,cell community eukaryotic,exogenous biodegradation and metabolism in YH group was significantly higher than that in M group(P<0.05).Conclusions: 1.Clinical study: In DKD patients,the deficiency of spleen and kidney was the main syndromes,and the standard evidence was mainly blood stasis syndrome and phlegm turbidity syndrome.2.Animal experiments: Yishenhuashi granules can regulate the intestinal flora structure through the “gut-kidney axis”,reduce intestinal permeability,increase the diversity of flora,improve the abundance of lactobacillus and other probiotics,thus improving glucose and lipid metabolism,reducing inflammation,reducing urinary toxins in DKD rats,and improving renal function.