Study On The Mechanism Of Paidu Baoshen Pills In Treating Chronic Kidney Disease Rats Based On Intestinal Microecology And Its Mediated Inflammatio

Objective: Based on the theory of "gut-kidney axis",this study intends to take 5/6nephrectomy model rats as the research object,to explain the mechanism of action of Paidu Baoshen Pills in the treatment of chronic kidney disease,and to provide a scientific empirical basis for its clinical treatment.Methods: SPF SD rats were randomly divided into sham operation group and model group.Rats in the modeling group were used to establish a 5/6 nephrectomy model.After modeling,they were randomly divided into model group,Paidu Baoshen Pills group,and Niaoduqing Granules group(positive control group).The Paidu Baoshen pill group was given Paidu Baoshen pill(0.54g/kg),the Niaoduqing group was given Niaoduqing(1.35g/kg),the model group and the sham operation group were given equal volume of distilled water,once a day for 8 consecutive weeks.After the last administration,various indicators were detected:(1)renal function and 24-hour urine protein quantification.(2)HE and Masson staining of kidney pathology.(3)Enzyme-linked immunosorbent assay was used to detect serum inflammatory factors interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),fecal secretory immunoglobulin A(secretory immunoglobulin A,s Ig A)content.(4)The contents of rat cecum were collected to detect the diversity of rat intestinal flora.(5)The rat intestine was taken for histopathological examination.(6)Rat kidney tissue was taken,and glutathione peroxidase 4(GPX4)was detected by immunohistochemistry;rat serum was taken,and glutathione(GSH)was detected by colorimetry Rat kidney tissue was collected,and reactive oxygen species(ROS)were detected by DCFH-DA fluorescent probe method.(7)The rat intestinal tissue was taken,and the Real-time PCR method was used to detect the toll like receptor 9(toll like receptor 9,TLR9)and nuclear factor kappa-B(NF-κB)in the rat intestinal tissue m RNA expression.Western Blot technology was used to detect the key protein molecules of TLR9/NF-κB signaling pathway TLR9,interferon regulatory factor 7(IRF7),tumor necrosis factor receptor-associated factor-6(tnf receptor-associated factor-6,TRF6),IRF7 and NF-κB p65 phosphorylation expression.Result:(1)Effects of Paidu Baoshen Pills on renal function in ratsCompared with the sham operation group,the serum creatinine,blood urea nitrogen and urine protein of the model group were significantly increased(P<0.05).Compared with the model group,the serum creatinine,blood urea nitrogen and urine protein in the Paidu Baoshen pill group were significantly decreased(P<0.05).(2)The effect of Paidu Baoshen Pill on the pathological changes of rat kidney tissue Renal pathology,HE and Masson staining,compared with the sham operation group,the renal pathological interstitial inflammatory infiltration and collagen deposition in the model group were obvious.Compared with the model group,the Paidu Baoshen Pill group had a significant improvement.(3)Effects of Paidu Baoshen Pills on inflammatory factors IL-6 and TNF-α in ratsCompared with the sham operation group,s Ig A in the model group decreased significantly;IL-6 、 TNF-α Significantly increased(P<0.05).After treatment,compared with the model group,s Ig A in the Paidu Baoshen pill group was significantly higher and inflammatory factors were significantly lower(P<0.05).(4)Effects of Paidu Baoshen Pills on oxidative stress in rats Compared with the sham operation group,GPX4 and GSH in theCompared with the sham operation group,GPx4 and GSH in the model group decreased significantly,and ROS increased significantly(P<0.05).Compared with the model group,GPx4 and GSH in the Paidu Baoshen pill group increased significantly,while ROS decreased significantly(P<0.05).(5)Effects of Paidu Baoshen Pills on the diversity of intestinal flora in rats.Compared with the sham operation group,the abundance of conditional pathogenic bacteria in the model group was relatively increased;(6)Effects of Paidu Baoshen Pills on intestinal pathology in ratsIntestinal pathology HE staining showed that compared with the sham operation group,a large number of lymphocytes infiltrated the intestinal tissue of the model group.Compared with the model group,the Paidu Baoshen Pill group was significantly improved.(7)Effects of Paidu Baoshen Pills on TLR9 and NF-κB m RNA in ratsCompared with the sham operation group,the expression of TLR9 in the model group was significantly increased(P<0.05).After treatment,the expression of TLR9 in Paidu Baoshen Pill group was significantly decreased(P<0.05).Compared with the sham operation group,the expression levels of NF-κB m RNA in the model group and NF-κB were significantly increased(P<0.05).After treatment,the expression of NF-κB m RNA in Paidu Baoshen Pill group was significantly decreased(P<0.05)(8)Effects of Paidu Baoshen Pills on TLR9/NF-κB pathway-related proteins in ratsCompared with sham operation group,TLR9,p-IRF7,TRF6,p-NF-κ B p65,p-IRF7/IRF7,p-NF-κB p65/NF-κB p65、NF-κB p65(nuclear)/ NF-κB p65 in model groupwas significantly increased(P<0.05);After treatment,compared with the model group,in the Paidu Baoshen pill group,TLR9,p-IRF7,TRF6,p-NF-κ B p65 、P-IRF7/IRF7 、p-NF-κB p65/NF-κB p65 、NF-κB p65(nuclear)/ NF-κB p65 decreased significantly(P<0.05).Conclusion: Paidu Baoshen Pills may improve the intestinal microecology and intestinal mucosal barrier function in CKD rats,and then act on the TLR9/NF-κB pathway,inhibit the release of pro-inflammatory cytokines IL-6 and TNF-α,and inhibit oxidation.stress and exert its therapeutic effect.