| Objective:To observe the expression of TGF-1/Smad2 pathway and related factors in renal tissue of DN rats by LHSH,and to explore the possible mechanism of LHSH delaying renal fibrosis of DN.Materials and methods:80 SD rats were divided into control group(sham group),model group(DN),western medicine group(ARB)and LHSH compound treatment group(LH).LHSH compound treatment group was divided into LHSH compound optimization formula(LHD),LHSH compound high dose group(LHB)and LHSH compound low dose group(LHA).The right kidney of the rat was removed and fed with high sugar and fat.One month later,a 45mg/kg intraperitoneal injection of streptozotocin(STZ)was performed.When blood glucose reached16.7mmol/l and urine protein was positive,the model was successfully established.Sham group rats were given the same trauma except the right kidney was not removed,and then intraperitoneally injected with sodium citrate buffer 45mg/kg.After the successful modeling,normal saline of 1ml/d was given to sham group and DN group,ARB group was treated with irbesartan tablets at 75mg/kg/d,The LHD group was given LHSH compound to add and subtract group750mg/kg/d,The LHB group was given LHSH compound high dose group750mg/kg/d,The LHA group was given LHSH compound low dose group 375mg/kg/d.After 8weeks of administration,HE staining was used to observe renal histological changes,Masson staining was used to detect renal fibrosis,biochemical tests were used to detect renal function,blood glucose and blood lipids,routine urine tests were used to detect urinary ACR and24-hour proteinuria,ELISA was used to detect inflammatory factors,and Western Blot was used to detect the expression changes of NF-B and TGF-1/Smad2 pathways.Result:1.General situation: the rats in DN group had typical clinical symptoms such as polydipsia,polyphagia,polyuria and weight loss.LHSH compound had significant improvement on the mental state,activity and symptoms of diabetic nephropathy in rats.2.Biochemical test: compared to Sham group,serum creatinine,urea nitrogen and blood lipid levels were increased in DN group,it is statistical significance(p< 0.05).Both LH group and ARB group had a decreasing effect on the level of biochemical indexes in DN group.However,the therapeutic effect of LHD group was better than that of ARB group,and the difference was statistically significant(p < 0.05).3.Urine routine: compared to the DN group,the quantitative levels of urine ACR and 24-hour urine protein in the LH group and the ARB group were decreased,showing statistical difference(p < 0.05).The therapeutic effect of LHD group was significant compared with that of ARB group(p< 0.05).The therapeutic effect of LHA group and LHB group was lower than that of ARB group(p < 0.05),and there was no difference between the two groups.4.Histological examination: under light microscope,HE and Masson staining indicated that hypertrophy of glomerulus,thickening of basement membrane and proliferation of mesangial stroma were observed in rats in the DN group.Renal vesicles were fissure-like,and there was even balloon adhesion.There was no edema in tubule interstitium,granulosity of epithelial cells,turbidity and swelling,and no brush-like edge shedding.Inflammatory cells infiltrate the cortical stroma,and there is focal interstitial fibrosis.All the treatment groups had a remission effect on renal pathological changes,and the renal pathological changes in LHD group were less severe than those in other treatment groups.5.Inflammation detection: the expression of IL-18 was detected by ELISA,and the expression of Nox4 protein was detected by WB.The expression levels of IL-18,Nox4 and NF-?B were the highest in the DN group,and the lowest in the sham group(p < 0.05).Both LH and ARB groups inhibited the expression of IL-18,Nox4 and NF-? B.There was a statistically significant difference between the LHD group and the ARB group(p < 0.05),while there was no difference between the LHA group and the LHB group.6.Fibrosis index: no CTGF protein expression was observed in sham group kidney,while CTGF protein expression was observed in DN group kidney.The expression was strongest in the DN group(p < 0.05).Both LH group and ARB group had the effect of inhibiting theexpression of CTGF protein,and there was a difference between the two groups(p<0.05).Regarding the TGF-1/Smad2 signaling pathway and CTGF protein expression,compared to sham group,the TGF-1/Smad2 signaling pathway and CTGF protein expression were significantly up-regulated in the DN group,and the difference was statistically significant(p< 0.05).LHD group had an inhibitory effect on the up-regulation,which was significantly higher than that of ARB group(p< 0.05).Conclusion:1.LHSH(LHD)can improve the general situation of early diabetic nephropathy rats on renal function and reduce indicators such as urinary protein.It has a clear renal protective effect on diabetic nephropathy.2.LHSH(LHD)can inhibit the inflammatory response in early diabetic rats,and its mechanism is related to the inhibition of IL-18,Nox4 protein and NF-?B.3.LHSH(LHD)can alleviate renal fibrosis in early diabetic rats,and the TGF-1/Smad2 signaling pathway plays a major regulatory role.LH can alleviate renal fibrosis in early diabetic rats,and the TGF-1/Smad2 signaling pathway plays a major regulatory role. |
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