|
Objective: Overtraining-induced acute kidney injury (overtraining-induced acute kidney injury, OTIAKI) has become a major disease hazard to human health, the research attention in recent years, but the damage mode and pathogenesis is unclear, yet clinically lack of effective biomarkers for early diagnosis and prevention measures. Therefore, in-depth study OTIAKI pathogenesis and seek early diagnosis markers and is currently clinical medicine, sports medicine and important issue facing military medicine. Numerous studies show that kidney injury molecule -1 (kidney injury molecule-1, Kim-1) and interleukin -6 (Interleukin-6, IL-6) is the early diagnosis of AKI important biological indicators. In this study, rats were trained to swim to exhaustion caused by excessive build a model of acute kidney injury, overtraining was observed in rats Kim-1 and IL-6 in change and its relationship with kidney injury as well as the impact of anisodamine to deepen their understanding of the pathogenesis, seek new sensitive and specific markers for early diagnosis, enrich their control measures for the early diagnosis and provide the basis for improved treatment level. Methods: (1) Select 56 healthy male SD rats weighing 200-220g. The rats were randomly divided randomly into three groups: sedentary control group (control, CN, n = 8), exhaustive exercise group (exhaustive swimming, ES, n = 24), anisodamine group (Anisodamine, AD , n = 24). CN is the quiet control, did not carry out exhaustive exercise; ES group was based on recovery time after exhaustion 1h after exhaustive divided into groups (ES 1h), exhausted after 6h group (ES 6h) and 24 h after exhaustive group ( ES 24h); AD exhaustive exercise group at 20 minutes before the intraperitoneal injection of anisodamine (10mg/kg) after exhaustive exercise, divided into anisodamine 1h group (AD 1h), anisodamine 6h group (AD 6h) and anisodamine 24h group (AD 24h). Each time point were eight mice. Using non-weight-bearing rats swimming to exhaustion caused by overtraining established model of acute kidney injury. (2) the normal control group, exhaustive (6h, 24h) group, anisodamine (6h, 24h) after exhaustive exercise group was placed in metabolic cages to collect urine, the urine was measured by radioimmunoassay Kim- an activity. Blood from the inferior vena cava, were injected into ordinary test tubes and pipes used anticoagulant AU2700 automatic biochemical analyzer measuring serum creatinine (Scr), blood urea nitrogen (BUN) levels and enzyme-linked immunosorbent assay IL-6 activity. Production was cut into the side of the kidney homogenate stored at -20 ℃ by enzyme-linked immunosorbent assay Kim-1 and IL-6 activity; contralateral kidney placed in 10% neutral formalin fixed, for HE staining, immunohistochemistry Chemical slice to observe pathological changes in the kidney and Kim-1 expression. (3) Statistical analysis: All experimental data were analyzed by SPSS 13.0 software package for statistical analysis, the experimental data are presented as mean ± standard deviation (X - ± s) expressed. Among the groups were compared using ANOVA (one-way ANOVA) analysis, correlation analysis was performed using Pearson correlation test to p lt; 0.05 was considered statistically significant. Results: (1) normal control rats flexible operation, spiritual good, responsive, normal diet; rather exhaustive exercise group were limp and weak, apathetic, indifferent reaction, eating less; anisodamine situation better than rats same period of exhaustive exercise group. (2) normal renal tissue structure of the control group did not change. Exhausted Exhausted after 1h and 6h after slight changes in renal tissue structure, only see part of the glomerular capsule and tubular dilatation, vacuolar degeneration; Exhausion 24h group showed edema proximal tubular epithelial cells, membrane rupture , renal tubular epithelial cell brush border irregular, or even fall off. Anisodamine intervention group than in the same period of exhaustive exercise alone group pathological changes slightly. (3) 1h after exhaustive serum BUN, Scr that is significantly higher, 6h peaked, 24h returned to normal control levels; anisodamine 1 hour, 6 hours group BUN, Scr significantly lower than exhaustive exercise group (p lt; 0.05), but still higher than the normal control group (p lt; 0.05). (4) in plasma IL-6 activity in exhaustion significantly increased after 1 hour (p lt; 0.05), after exhaustive exercise 6h, 24h group compared with the control group had no significant difference (p gt; 0. 05) . Renal tissue IL-6 activity at 1 hour after the exhaustion was significantly higher (p lt; 0.01), exhaustive six hours gradually reduced (p lt; 0.05), until exhaustion 24 hours returned to normal control levels. Anisodamine 1h, 6h renal tissue IL-6 activity than simply exhausted group (p lt; 0.05), with no difference compared with the control group (p gt; 0. 05). (5) Kim-1 in urine after 6 hours at exhaustion was increased (p lt; 0.05), until exhaustion reached a peak after 24 hours (p lt; 0.01); Anisodamine the renal tissue Kim- an activity lower than that exhaustive exercise group (p lt; 0.05), with no difference compared with the control group (p gt; 0.05). Exhausted after renal tissue Kim-1 activity increases recovery time after exhaustion increased gradually, the exhausted group compared with the control group were statistically significant (p lt; 0.05). Anisodamine 1h, 6h group Kim-1 activity than simply exhausted group (p lt; 0.05), with no difference compared with the control group (p gt; 0.05); anisodamine renal tissue Kim-24h 1 activity was also lower than that exhausted group (p lt; 0.05), still higher than the normal control group (p lt; 0.05). (6) the normal control group only weakly expressed Kim-1, exhausted after one hour compared with the control group, the expression of enhanced (p lt; 0.05), exhaustive exercise 6 hours and 24 hours after the expression continues to strengthen, Kim anisodamine intervention group 1 expression simply exhausted group compared with the same period decreased significantly (p lt; 0.05), but still stronger than the normal control group. (7) The correlation analysis: Plasma IL-6 and renal tissue IL-6 activity was positively correlated (r = 0.75, p lt; 0.01); urinary Kim-1 and renal tissue Kim-1 activity was positively correlated (r = 0.632 , p lt; 0.05); urinary Kim-1 expression and Scr levels were significantly correlated (r = 0.932, p lt; 0.01). Conclusion: a renal tissue Kim-1 activity at 1h after exhaustive exercise significantly increased urinary Kim-1 activity at 6h after exhaustion was significantly increased, suggesting that Kim-1 may be OTIAKI early biological indicators;, and may vary injury time extended gradually increased, suggesting that Kim-1 activity in a time dependent manner, and with the severity of injury and related developments. 2 IL-6 activity in exhaustive exercise one hour that is significantly increased, and then gradually decreased to normal levels, suggesting that IL-6 may be overtraining induced acute kidney injury early play an important role. 3 anisodamine can significantly reduce the Kim-1 and IL-6 activity in rats on OTIAKI play a significant protective effect, reduce kidney damage caused by overtraining.
|