| Purpose:In this study,the expressions of P53,SLC7A11,GPX4,COX2,NOX1 and FTH1 in Diabetic Kidney Disease(DKD)model rats were observed,and the regulation of P53,SLC7A11,GPX4,COX2 and NOX1 by Chinese traditional medicine compound Yitangkang was explored.Effect and mechanism of FTH1 inhibiting iron death on kidney injury in DKD rats.Materialandmethod : Fifty SPF healthy Wistar rats were randomly divided into 5groups by random number table method,including blank control group,model control group,Yitangkang low-dose group,Yitangkang medium-dose group and Yitangkang high-dose group,with 10 rats in each group.After 4 weeks of feeding,the diabetic nephropathy animal model was replicated,and streptozotocin(STZ)was intraperitoneally injected at 45mg/kg in all groups.After successful modeling,mice in the three groups were given low,medium and high dose intragastric administration of yitangkang decoction at low,medium and high dose,respectively.Model control group and blank group were given equal volume of normal saline intragastric administration once a day,for a total time of 6 weeks.Determination of dose is calculated by using the conversion formula of human and biological crude drugs.The dosage of traditional Chinese medicine is the clinical dosage of Yitangkang.The dosage of high dose is equal to two times of the middle dose,and the dosage of low dose is equal to 0.5 times of the middle dose.In other words,rats in yitangkang medium-dose,high-dose and low-dose groups were given the same volume of Yitangkang decoction 10.5g/kg,21g/kg and 5.25g/kg by gavage,respectively.Fasting blood glucose and body weight of rats were detected before modeling and at the second,fourth and sixth weekend of Yitangkang intervention,and samples were taken at the sixth weekend after treatment.The levels of urinary protein,serum creatinine and urea nitrogen were detected by automatic biochemical analyzer.At the same time,the levels of MDA,T-SOD and GSH were detected by Elisa.The histopathological changes,especially the changes of glomerulus and renal tubules,were observed by HE staining.The expression of P53,GPX4,FTH1,SLC7A11,COX2 and NOX1 proteins and gene expression in kidney tissues of rats were detected by Western Blot and RT-PCR.Results:1.Morphological level: the blank group rats have a good mental state,flexible movement,moderate water intake,good eating,normal urinary excretion,smooth coat color,good gloss;compared with the blank group,the weight of the rats in the model group has dropped sharply after STZ intraperitoneal injection,and at the same time,the amount of water and urine excreted have also increased significantly,and the mental aspect has shown a sluggish state,fatigue,slow movement,and poor nutrition;after intervening with the low,medium and high dose groups of Yi Sugar Kang,it is lower than that of the model group.The mental state of the rats in the medium and high dose groups began to improve,the weight showed a recovery trend,the food intake improved,the water intake decreased,and the symptoms of urinating frequency also improved significantly.2.Body weight: compared with the blank group,the body weight of the remaining four groups was significantly increased after feeding the high-fat diet(P < 0.01);And the chain was produced by intraperitoneal injection of urea with cephalosporins quantitative after the rat model of diabetic nephropathy,compared with the blank control group and model control group,and yi sugar,low,medium and high dose group,the rats’ weight were present at the time of two weeks to detect decline(P < 0.01),compared with the four groups of mice weight between groups was no significant difference(P < 0.01);After intragastric administration of Yitangkang,compared with the blank control group,the body weight level of the other four groups was still lower than the blank control group(P < 0.05 or P < 0.01);Compared with model control group,body weight in low,medium and high dose Yitangkang groups was significantly increased(P < 0.01);Compared with low-dose and medium-dose groups,the body weight of yitangkang high-dose group was higher but the difference was not significant(P > 0.05).3.Fasting blood glucose level: after modeling,compared with blank control group,the fasting blood glucose level of model control group,yitangkang low,medium and high groups showed a trend of significant increase(P < 0.01),and there was no significant difference in fasting blood glucose level between these four groups(P > 0.05);After 2,4,and 6 weeks of intragastric administration of Yitangkang,the fasting blood glucose level of rats in TCM group 3 was slightly higher than that in blank control group(P < 0.05);Compared with model control group,yitangkang low-dose,medium-dose and high-dose groups showed a significant decrease in fasting blood glucose level(P < 0.01);Compared with the low-dose group,the fasting blood glucose level of yitangkang medium-dose and high-dose groups was decreased more significantly(P < 0.05).4.Renal function level: After the test,the content of blood creatinine,blood urea nitrogen and urine protein in the model group was significantly increased compared with that in the blank group(P<0.01);compared with the model control group,it was found that the blood creatinine,blood urea nitrogen and urine protein content of the three groups of Chinese medicine Yi sugar kang were significantly reduced(P<0.01);compared with the low dose group,the urine blood creatinine,blood urea nitrogen and urine protein content of rats in the middle and high dose groups of Yi sugar kang were more significantly reduced(P<0.05)..5.Renal pathology: rats in the blank group have normal glomerular size,morphological order,no inflammatory infiltration of the renal interstitium,no visible hyperplasia in the mesangial region,neatly arranged renal tubules,and complete structure;compared with the blank group,the number of kidney cells in the model group is significantly reduced,the glomeruli are significantly increased,the vacuoles are present in the cells of the renal tubular epithelium,the mesangial cells appear to proliferate significantly,and the mesangial matrix also increases significantly;compared with the model control group,the size and morphology of the glomeruli of rat rats in the low,medium and high dose groups of Yi Sugar Kang tend to be normal At the same time,the arrangement of renal tubules also tends to be neat,and the structural relatively complete basement membrane hyperplasia is slight,which is significantly less than that of the model group.6.Oxidative stress levels: The T-SOD and GSH levels in the model group were significantly reduced compared with those in the blank control group,but the MDA content showed a significant increase trend(P<0.01);compared with the model control group,the levels of T-SOD and GSH in the low,medium and high dose groups of Yixinkang were significantly improved,while the levels of MDA showed a significant decrease(P<0.05,P<0.01);compared between the low,medium and high doses of Yishankang in the three groups,the increase in T-SOD and GSH levels in the medium and high dose groups was the most significant,and the decrease in MDA levels was also the most significant(P<0.05 or P<0.01);there was no significant difference between the medium and high doses of Yixinkang(P>0.05).7.Iron death-related factor protein and m RNA expression: Compared with the blank control group,the expression levels of P53,COX2 and NOX1 in the model group were significantly increased,while the expression levels of SLC7A11,GPX4 and FTH1 were significantly reduced(P<0.05,P<0.01);the expression levels of P53,COX2 and NOX1 were significantly reduced in the low,medium and high dose groups of Yishankang compared with the model control group,while the expression levels of SLC7A11,GPX4 and FTH1 were significantly increased(P < 0.01).The difference between the low,medium and high doses of Yishankang was not obvious(P > 0.05).Conclusion:1.TCM Compound Yitangkang has an outstanding effect on regulating the disorder of glucose metabolism and the level of renal function in diabetic nephropathy rats in a dose-dependent manner.2.TCM compound Yitangkang has a dose-dependent effect on recovering the pathological damage of renal tissue,especially the damage of renal tubules in diabetic nephropathy rats.3.Yitangkang can activate the expressions of SLC7A11,GPX4 and FTH1,and inhibit the expressions of P53,COX2 and NOX1.4.The Traditional Chinese medicine Compound Yitangkang can regulate the kidney of DKD rats by affecting iron death,and its specific mechanism is realized by activating the expressions of SLC7A11 and GPX4. |
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