| Diabetic Nephropathy is one of the most serious complications of diabetes, which greatly affects the quality of life of the patients. The main pathological features of Diabetic Nephropathy are glomerular hypertrophy, tubular thickening and mesangial matrix accumulation. With the economic development, people’s living standards have improved and diet structure has changed. As a result, the incidence of diabetes and diabetic nephropathy has gradually increased. However, the pathogenesis of Diabetic Nephropathy has not been fully elucidated by Western Medicinal research, so the treatment of Diabetic Nephropathy is still on the way. Clinical studies have shown that Traditional Chinese Medicine (TCM) Radix Astragali and Radix Puerariae Lobatae has a certain therapeutic effect on diabetic nephropathy, so it is important to deeply study the therapeutic mechanism of Radix Astragali and Radix Puerariae Lobatae on Diabetic Nephropathy. Endoplasmic reticulum stress is one of the focus at home and abroad, and there is a certain association between the endoplasmic reticulum stress and Diabetic Nephropathy. Therefore, it is significant to conduct deep study of the mechanism of endoplasmic reticulum stress in Diabetic Nephropathy by Traditional Chinese Medicine Radix Astragali and Radix Puerariae Lobatae.Objective:Analyzing the influence of the changes of XBP1, related protein and mRNA of the endoplasmic reticulum stress in the treatment of the early stage of Diabetic Nephropathy to discuss the treatment and mechanism of the injection of Puerarian and Astragalus.Method:1. Divide the SPF level diabetes model mice KKAy into model group and treatment group, the same age mice C57BL/6J as the control group. The treatment group were administered by intraperitoneal injection from the12weeks. Weighted the body weight of mice every other week, measured the blood glucose at12,14,16weeks. Specimen collection in16th week, detect the biochemical indexes and morphological changes and RNA, protein expression.2. Divide the Human renal tubular epithelial cells (HK-2cells) into control group, model group and treatment group. Add culture medium with high glucose after of adherent in model group, detect the expression of mRNA and proteins after adding Astragalus injection combined puerarin injection in24h,48h,72h.Result:1.The KKAy mice occurred a significant obesity, food increased, water intake, and high blood sugar. Compared with the control group, the blood sugar of treatment group was lower than the model group in16th week, the renal function of treatment group was improved, and pathological changes of treatment group also significantly reduced.The expression of GRP78mRNA in model group was significantly higher than the normal group(P<0.05), the expression of GRP78mRNA and proteins in treatment group were lower than the model group(P<0.05). The expression of XBPls mRNA and proteins in model group and treatment group were higher than the control group(P<0.05), the expression of protein of IRE1-P in model group and treatment group were higher than the control group(P<0.01), and the expression of IRE1-P in model group were higher than the model group(P<0.01). the expression of XBPls mRNA in treatment group was higher than the model group, the expression of protein of XBP1was lower than the model group, the expression of phosphorylated proteins of IRE1was higher than the model group.2. In the detection of three points in time, there is no statistical differences between groups in the time24hours. The expression of XBPls mRNA in treatment group and model group were higher than the control group in the time48and72hours (P<0.05). The mRNA expression of XBPls and GRP78in model group were higher than the control group (P<0.05). While the treatment group was lower than the model group (P<0.05). In the expression of proteins by Western Blot detection, the expression of GRP78ã€IRE1ã€XBP1were higher than the control group in72hours(P<0.05). The expression of GRP78in treatment group was lower than the model group(P<0.05). The expression of IRE1in treatment group was higher than the control group (P<0.01), The expression of XBP1in treatment was higher than the control group (P<0.05).Conclusion:1. The combination of Astragalus injection and Puerarin injection have a therapeutic effect in the early stage of DN, which mainly reflected in the reduce of blood sugar, improvement of renal function, and the reduce of kidneys disease.2. There is ERS reaction in the early stage of DN, in the expression of mRNA and protains, the main reaction was the synthesis of XBP1smRNA.3. The combination therapy can regulate the ERS reaction in the early stage, which adjust the pathway of XBPls, to protect the tubular epithelial cells and the kidney function. |
