| Objective:We investigate the localization and expression of Integrin Linked Kinase(ILK) in rats by Unilateral Ureteral Obstruction (UUO) model and the role of ILK in Tubularinterstitial Fibrosis(TIF), in order to explore the effect of Centella asiatica on ILK and ILK-mRNA expression, to clarify the mechanisms of Centella asiatica in easing TIF and provide theoretical basis for the clinical application of centella asiatica in preventing TIF.Method:The method of ligating the left ureter were used in establishing the model.60 male SD rats were randomly divided into control group, model group, high-dose Centella asiatica (HC) group, intermediate-dose Centella asiatica (MC) group, low-dose Centella asiatica (LC) group, Fosinopril group,10 rats in each group. UUO rat model were set up except the control group. Treatment groups were given a gavage of appropriate doses while control group and model group were given same amount of saline.14 days after the operation, all rats were executed for the following tests:1, Blood were taken from the abdominal aorta for the test of SCR and BUN.2, The obstruction side kidney tissue were taken at the same time for the use of HE staining in order to observe the pathological changes, tubular dilatation and atrophy of obstructed kidney; Masson staining for the evaluation of renal interstitial fibrosis; immunohistochemical staining for the detection of localization and expression tendency of ILK protein in renal tissues.3, The technology real-time PCR (rt-PCR) were used to detect tissue ILK-mRNA levels.Results:1, Rats' postoperative conditions:one model group rat died 2 days after the operation, one LC group rat died 4 days after the operation. The rats'wound healed well 5 days after the surgery. UUO model rats'activity and fur lustrousness decreased compared with control group.2, UUO groups saw increased SCR and BUN levels compared with the control group, with a significant difference (P<0.05), there is no statistical difference between UUO groups among two kidney function indicators(P> 0.05).3, Conventional HE and Masson staining:control group saw no significant pathological changes; UUO group saw significant tubular expansion of various levels, part of small tubular atrophy, large amount of interstitial inflammatory cell infiltration and small amount of fiber leakage and increased collagen fibers after 14 days. Treatment group was lower than model group but still higher than control group (P<0.05).4, Results of immunohistochemical methods in detecting localization and expression of renal ILK protein and RT-PCR methods in detecting ILK-mRNA main ILK expression was in renal tubular epithelial cells, with a small amount of expression within the nucleus, and a small amount of basal expression in glomerular endothelial cells and vascular endothelial cells; control group saw no significant difference (P> 0.05), ILK protein and mRNA levels in each UUO group were higher than that of the control group. Within UUO groups, the expression in those groups with drug intervention was lower than the model group (P< 0.05); LC group was similar with the MC group (P> 0.05) and HC group was almost the same as Fosinopril group (P> 0.05). The expression in HC group and Fosinopril group were lower than that in LC group and MC group (P< 0.05). There was no statistically significant difference of SCR and BUN between those drug intervention groups(P> 0.05).Conclusion:1, ILK protein and ILK mRNA level is obviously higher in UUO rats'renal tissue than in normal conditions, indicating that ILK plays an important role in the process of TIF.2, Centella asiatica could probably alleviate TIF by regulating the expression of ILK and promoting the Extracellular Matrix (ECM) degradation. The effect of Centella asiaticais is highly related to its dosage, and effect of high-dose Centella asiaticais is the equivalent of Fosinopril.3, It is safe to use Centella asiaticais for with no obvious renal toxicity. |
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