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Purpose established by intraperitoneal injection of endotoxin (lipopolysaccharide, LPS) in rats with acute kidney injury (Acute kidney injury, AKI) model to explore fasudil hydrochloride (Fasudil hydrochloride, Fasudil) AKI serum tumor necrosis factor-alpha (Tumor necrosis factor-alpha, TNF-α), endothelin -1 (Endothelin-1, ET-1), renal tissue Rho kinase - Ⅰ (Rho kinases-Ⅰ, ROCK-Ⅰ) and nuclear factor-kappa B p65 (Nuclear Transcription Factor of Kappa B p65, NF-kappaB p65) the impact of changes, renal protective effect of Fasudil in AKI. Methods Male clean grade 65 Wister rats were randomly divided into control group, the AKI model group and treatment group. Control group: 5. 1 ml of saline by intraperitoneal injection. Model group: 30. (1h, 6h, 12h, 24h, 48h, 72h), at each time point for each five rats were randomly divided into six time points. LPS, 6mg/kg body weight, diluted with normal saline to 1 ml intraperitoneal injection of AKI model. Treatment groups: 30. (1h, 6h, 12h, 24h, 48h, 72h), at each time point for each five rats were randomly divided into six time points. Each group is given before the injection of LPS the Fasudil 30mg/kg body weight, the same saline diluted to 1 ml intraperitoneal injection. 48h and 72h 24h additional the Fasudil 30mg/kg. Observed in each group, depending on the point in time ranging from 1 h-72h, 5 ml of blood in the corresponding time points after the rats were killed. Determination of endotoxin levels in plasma, serum creatinine (Cretonnes CRE), blood urea nitrogen (blood urea Nitrogen, BUN), ELISA (enzyme-linked immunosorbent assay, ELISA) detection of serum TNF-alpha, ET-1 the concentration of content. Immunohistochemistry was used to detect renal tissue ROCK-I, NF-kappaB p65 expression. The optical microscope synchronization observed each rat renal pathological changes. 1 of serum CRE and BUN change comparison: model group CRE, BUN 6h significantly increased compared with the control group, a statistically significant difference (P lt; 0.05); and peaked at 24h, 48h then decreased significantly. Treatment group at each time point CRE, BUN changes than model group corresponding time points increased significantly model 12h the 24h group CRE.BUN and treatment of 12h, 24h group, a significant difference (P lt; 0.01). (2) changes in serum TNF-alpha: the levels of serum TNF-alpha concentration immediately 1h significantly increased compared with the control group, there were significant differences (P lt; 0.01), and then decreased rapidly 6h and its later point in time has been in a relatively low level, single peak curve was steep; TNF-alpha concentration changes in the treatment group at each time point than model group was significantly 1h group model of TNF-alpha concentration hour treatment group compared to the statistically significant difference ( P lt; 0.05). Serum ET-1 changes: the levels of serum ET-1 concentration was significantly increased at 1 h, a significant difference compared with the control group (P lt; 0.01), and then began to decline and reached the peak at 12h, 72h basic to normal. Model 6h, 12h, 24h group ET-1 change with treatment group corresponding time points were significantly different (P lt; 0.05) (4) changes in plasma LPS: the model group and treatment group, plasma LPS level in 1h rapid increase, compared with the control group, a significant difference (P lt; 0.01), and peaked at 6h, 12h and rapidly decreased in the model group corresponding time points compared with the treatment group were not statistically different. 5 Comparison of changes in renal tissue ROCK-I expression: control group ROCK-I in renal tubular epithelial cells display trace expression; model 1h group Rock-I positive area was significantly increased compared with the control group, the difference was statistically significance (P lt; 0.05); and reached the peak at 12h, 24h began to decline. Model 6h, 12h, 24h, 48h group Rock-I expression area and the treatment group corresponding time points were significantly different (P lt; 0.05) 6. Comparison of changes in renal tissue NF-kappaB p65 expression: control group, NF-κB p65 in glomerular and tubular micro-expression. Model group, NF-kappaB p65 positive area in an immediate increase in 1h, compared with the control group, a significant difference (P lt; 0.01), and reached the peak at 12h, 24h group decreased significantly. Model 1h, 6h, 12h, 24h set of positive area and the treatment group corresponding time points were significantly different (P lt; 0.05) Biopsy generally normal control: model group early glomerular renal tubular epithelial cell swelling, fatty degeneration and vacuolar degeneration, late emergence of varying degrees of renal tubular epithelial cell necrosis, nuclear condensation, dissolved and disappeared. Necrosis of the renal tubular epithelium into the lumen, was a blur granular structure. Tubules interstitial congestion, edema and inflammatory cell infiltration. The treatment group had renal tubular epithelial cell swelling, a small amount of degeneration and necrosis, no significant changes in renal interstitial and inflammatory cell infiltration. 8 tubular necrosis Rating: model group at the 1h severe renal pathological damage, and peaked at 24h. Model 1h tubular necrosis score was significantly higher significant difference (P lt; 0.01), compared with the control group. Compared to the model group at all time points tubular necrosis score and treatment group were statistically significant (P lt; 0.05) Conclusion Fasudil may be blocking Rho / Rho kinase pathway and NF-kappaB p65 pathway, reducing ET-1 and inflammatory cytokines TNF-alpha expression level, to improve renal function, to reduce the LPS-induced renal injury, has a protective effect on the LPS-induced rat kidney AKI.
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