| Objective: To study the self-styled KKAy mice with Diabetic nephropathy as the research object,and observe the general condition and pharmacodynamics of diabetic nephropathy mice treated with Baxiancao Decoction,with irbesartan and metformin as drug control groups,and pathological morphology.Study,immunohistochemistry,Western Blot,real-time fluorescent quantitative PCR and other experimental techniques to observe the expression of NLRP3,Casepase-1,IL-1β inflammatory factors protein and gene in the kidney of early diabetic nephropathy KKAy mice The protective mechanism of decoction on the kidney of early Diabetic nephropathy and its effect on NLRP3-Casepase-1-IL-1βsignal transduction pathway provide an experimental basis for the antagonistic inflammatory injury of Baxiancao Decoction.Methods:1.Effect of Baxiancao Decoction on the pharmacodynamics of Diabetic nephropathy KKAy miceSeventy KKAy female mice and 10 C57BL/6J female mice at 7 weeks of age were purchased.After 1 week of free feeding and drinking,they were given KKAy mouse-specific feed for 4 weeks.When KKAy mice reached blood glucose ≥ 16.7 mmol/ L;24h urine protein ≥ 0.4mg mice successfully modeled for diabetic nephropathy.C57BL/6J mice were randomly divided into blank groups,and the rest were randomly divided into model group,Baxiancao decoction low dose group,Baxiancao decoction high dose group,Baxiancao decoction plus irbesartan group,Baxiancao decoction plus metformin group,and erbe.Shatan group,metformin group.According to the model weight conversion,the corresponding group of drugs is administered by gavage.Animal weight,water intake,urine volume and general state were observed during the experiment.After 8 weeks of drug intervention,blood glucose was detected in the tail vein blood.Blood samples were collected from the blood to determine serum creatinine,serum urea nitrogen,total cholesterol,triglycerides,low density lipoprotein cholesterol,and high density lipoprotein cholesterol.Urine presentationused to detect urine protein and creatinine.The kidney weight of the mice was recorded and the kidney weight was calculated.Body weight ratio,while preserving kidney tissue according to the requirements of each experiment.2.HE staining and PAS staining were used to observe the pathological changes of renal tissue after treatment with Baxiancao Decoction.3.Immunohistochemistry was used to detect the expression of NLRP3 and IL-1β inflammatory factors in renal tissues.4.Western Blot was used to detect the quantitative expression of NLRP3,Casepase-1 and IL-1β inflammatory factors in renal tissues.5.Real-time quantitative PCR was used to detect the relative quantitative gene expression of NLRP3 and IL-1β inflammatory factors in renal tissues.Results:1.Effects of Baxiancao Decoction on the pharmacodynamics of Diabetic nephropathy KKAy miceCompared with the model group,the Chinese medicine group and the Chinese medicine plus western medicine can improve the activity of the mice,and the spirit is better than the model group.The hair loss and facial alopecia are less than the model group,and there are no symptoms such as white turbidity in the eyes and white turbidity in the urethra.Compared with the model group,the difference in the relative reduction of drinking water in the 5 weeks before treatment was statistically significant(P<0.05).Compared with the model group,the weight ratio of kidney to body weight of Chinese medicine plus irbesartan group and Chinese medicine plus metformin group was better than that of high dose group of traditional Chinese medicine(P<0.05).Compared with the model group,the blood glucose of the Chinese medicine plus metformin group decreased,the difference was statistically significant(P<0.05).Compared with the model group,the total cholesterol,triglyceride,low-density lipoprotein cholesterol,and high-density lipoprotein cholesterol of the high-dose Chinese medicine group,the Chinese medicine plus irbesartan group,and the Chinese medicine plus metformin group decreased,the difference was statistically significant.(P<0.05).Compared with the model group,the blood urea nitrogen and serum creatinine of the high-dose Chinese medicine,Chinese medicine plus irbesartan group and irbesartan group were significantly different(P<0.05).Compared with the model group,the 24-hour urine protein decreased in the high-dose Chinese medicine group,the Chinese medicine plus irbesartan group,and the Chinese medicine plus metformin group,the difference was statistically significant(P<0.05).2.HE and PAS results showed that compared with the model group,the glomerular capillary basement membrane was slightly thickened after treatment with high-dose Chinese medicine;mesangial matrix mild hyperplasia,mesangial cell mild hyperplasia,renal tubular epithelial cells Granular degeneration;focal infiltration of focal macrophages was observed in the renal interstitial.3.The results of immunohistochemistry showed that compared with the model group,the average optical density and positive area ratio of NLRP3 and IL-1β in the high-dose group of traditional Chinese medicine,Chinese medicine plus irbesartan group,and Chinese medicine plus metformin group decreased.Academic significance(P<0.05).4.Western Blot results showed that compared with the model group,the expression of NLRP3,Caspase-1 and IL-1β protein in the normal group was less,and the difference was statistically significant(P<0.05).Compared with the model group,the relative quantitation of NLRP3,Caspase-1 and IL-1βin the high-dose group,the Chinese medicine plus irbesartan group and the Chinese medicine plus metformin group decreased,the difference was statistically significant(P<0.05).5.Real-time quantitative PCR results showed that compared with the model group,the expression of NLRP3 and IL-1β in the normal group was less,and the difference was statistically significant(P<0.05).Compared with the model group,the gene expressions of NLRP3 and IL-1β in the high-dose group of Chinese medicine,Chinese medicine plus irbesartan group,and Chinese medicine plus metformin group decreased,the difference was statistically significant(P<0.05).Conclusions:1.Baxiancao Decoction can alleviate the symptoms of poor dementia and alopecia areata in KKAy mice,which can reduce the serum creatinine,blood urea nitrogen,triglyceride,total cholesterol,24-hour urine protein and other indicators in KKAy mice.Can slow down renal function damage in Diabetic nephropathy KKAy mice.2.Inflammatory damage plays an important role in the progression of Diabetic nephropathy,and inflammatory molecules such as NLRP3,Caspase-1,and IL-1β are involved in this inflammatory process.3.Baxiancao Decoction can improve the renal pathological structure of Diabetic nephropathy KKAy mice,and reduce the relative quantitative expression of NLRP3 and IL-1β in renal tissue.4.Baxiancao Decoction can reduce the relative quantitative protein expression of NLRP3,Caspase-1 and IL-1 β in renal tissue of Diabetic nephropathy KKAy mice,and reduce the relative gene expression of NLRP3 and IL-1β.5.Baxiancao Decoction has a certain protective effect on the kidney of Diabetic nephropathy KKAy mice.The mechanism of its existence may reduce the relative quantitative expression of NLRP3 and IL-1 β in renal tissue,and down-regulate NLRP3,Caspase-1,IL-1 β.The relative quantitative expression of relative quantitative protein and m RNA regulates the NLRP3-Caspase-1-IL-1 β signal transduction pathway,thereby slowing the inflammatory injury of diabetic nephropathy. |
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