| Tubular interstitial fibrosis (TIF), almost the common process of all kinds of chronic renal diseases progressing to end-stage renal failure, is characterized by tubular ectasia or atrophy, infiltration of inflammatory cells, proliferation or activation of fibroblast and deposition of extracellular matrix (ECM) in the pathological changes. Inflammatory reaction is not only one of the tumor-initiating links but also throughout the renal tubular interstitial fibrosis. Effectively inhibiting the inflammatory reaction plays an important part in controlling the occurrence and development of renal tubular interstitial fibrosis.Objective:Animal model with TIF made by unilateral ureteral obstruction (UUO) with valsartan as control drug, the study is to observe the effects of Jiedu Huoxue therapy on pathomorphology in renal tissue, serum creatinine (Scr), blood urea nitrogen (BUN), interleukin-1β (IL-1β), tumour necrosis factor-a (TNF-α), monocyte chemoattractant protein-1(MCP-1), and to explore the protective effect and possible mechanism of Jiedu Huoxue therapy in UUO rats, so as to provide experimental evidence for clinical treatment.Methods:Fifty healthy female Sprague Dawley (SD) rats were randomly divided into five groups:Control group, Model group, Valsartan group, Jiedu Huoxue decoction low-dose group and high-dose group. All animals were adaptively fed for one week. Except the Control group, rats in other four groups were made into unilateral ureteral obstruction (UUO) model by ligaturing the left ureter on the eighth day. All rats were treated with drugs by oral infusion once a day. Rats in Valsartan group, Jiedu Huoxue decoction low-dose group and high-dose group were treated respectively with Valsartan26mg/kg-d, Jiedu Huoxue decoction12g/kg-d and24g/kg-d for fourteen days and the volume was lml/l00g. Physiological saline of the same volume was given to rats in Control group and Model group, once a day for14days. All the animals were sacrificed on the twenty-second day. The specimen of serum was produced to detect the contents of blood urea nitrogen and serum creatinine. Take the left kidney and prepare the nephridial tissue bomogenate to detect IL-1β by radioimmunoassay, to observe the change of pathomorphology by HE and Masson staining, and to detect TNF-α and MCP-1by immunohistochemistry.Results:1. General StatesThere were no obvious changes on fur, diet, spirit, activity in rats in control group. The rats in Model group were listless, short of diet and activity, and its fur turned disheveled. The rats in other treatment groups were superior to those in Model group.2. Detection of the Contents of Serum Creatinine and Blood Urea NitrogenThe contents of serum creatinine (117.090±14.521μmol/L) and blood urea nitrogen (9.689±0.570mmo1/L) in Model group were significantly higher than those in control group (61.118±12.492,7.485±0.354)(P<0.01). The contents of serum creatinine (95.218±15.805,81.914±9.598,83.863±8.716) and blood urea nitrogen (8.832±0.289,8.374±0.297,8.573±0.344) in Valsartan group, Jiedu Huoxue decoction high-dose group and Jiedu Huoxue decoction low-dose group were obviously lower than those in Model group (P<0.0l).3. Changes of Pathomorphology in Renal TissueHE staining showed that the epithelial cells in renal tubule of control group arranged in neat rows without abnormal changes. There were degeneration, necrosis and abscission of the epithelial cells, tubular ectasia, and a large number of inflammatory cells infiltrating in interstitial space in renal tubule of Model group. In Valsartan group, Jiedu Huoxue decoction low-dose group and high-dose group were better than Model group.Masson staining showed that there were a few expressions of collagen in interstitial space in control group. But a lot of collagen deposited in interstitial space in Model group. Compared with Model group, the expression of collagen reduced significantly in Valsartan group, Jiedu Huoxue decoction low-dose group and Jiedu Huoxue decoction high-dose group.4. Expressions of TNF-α in Kidney Detected by ImmunohistochemistryThere were weak expressions of TNF-α in tubular epithelial cells in control group, pale yellow. Compared with control group, the expressions remarkably increased in Model group, brown orange. Compared with Model group, the expressions reduced in Valsartan group, pale yellow. Those in Jiedu Huoxue decoction low-dose group and Jiedu Huoxue decoction high-dose group obviously decreased, pale yellow.5. Expressions of MCP-1in Kidney Detected by ImmunohistochemistryThe expressions of MCP-1showed a small amount in control group. Compared with control group, the expressions significantly increased in tubular epithelial cells in Model group, brown orange. Compared with Model group, the expressions decreased in Valsartan group, pale yellow. Those in Jiedu Huoxue decoction low-dose group and Jiedu Huoxue decoction high-dose group remarkably reduced.6. Expressions of IL-1β in Kidney Detected by RadioimmunoassayThe expressions of IL-1β in Model group (144.510±14.462pg/ml) were remarkably higher than those in control group (107.640±14.657)(P<0.01). Compared with Model group, they sharply decreased in Valsartan group (130.590±9.803), Jiedu Huoxue decoction low-dose group (127.420±11.274) and Jiedu Huoxue decoction high-dose group (124.580±12.060)(P <0.01).Conclusions:1. Jiedu Huoxue decoction works to reduce significantly the injury of the structure of renal tissue in TIF rats and improve renal function. It is confirmed that Jiedu Huoxue decoction plays a role in decreasing obviously renal tubular interstitial fibrosis and protecting the kidney from the experimental angle.2. The high expressions of inflammatory cytokines IL-1β, TNF-α, MCP-1 in renal tissue in unilateral ureteral ligation rats show that inflammatory reaction is one of the important mechanisms leading to renal tubular interstitial fibrosis.3. Jiedu Huoxue decoction is able to reduce the high expressions of IL-lβ, TNF-α, MCP-1in renal tissue in unilateral ureteral ligation rats, and to retard the progress of renal tubular interstitial fibrosis by inhibiting inflammatory reaction. |
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