Expression Of TLR4and Fas And The Protective Effects Of Anisodamine In Exhaustive Swimming Rats

Objective: Our previous researches suggest that renal tubular epithelial cell apoptosis isthe main pathological characteristic of over-training-induced acute kidney injury (OTIAKI)and Anisodamine can relieve renal damage by inhibiting excessive renal tissue cellapoptosis,but the exact molecular mechanisms are still unclear. Toll-like receptors（TLRs）are well known as Pathogens in the innate immune system aimed as defending the survivalof the host and play critical roles in tissue injuries and functional deficits. Toll-like receptor4(TLR4),a key immune recptor in the TLRs family,has been considered “a injury-sensitiveprotein” and express highly to induce inflammation and apoptosis in acute kidney injurycaused by many reasons.In the present study,we established the animal model ofover-training-induced acute kidney injury by exhaustive swimming to observe theexpression changes of TLR4,Fas and cell apoptosis,to evalue the relationship between therenal tissue cell apoptosis and TLR4and investigate the influences of Anisodamine onabove changes.Methods:(1) Select48healthy SD rats, weighing200-220g,were randomly divided intothree groups: control group (CN), exhaustive swimming group (ES) and Anisodaminegroup (AD).(2)The animal model of overtraining-induced acute kidney injury wasdeveloped by exhaustive swimming.(3) The renals of rats were obtained for determinationof expression of TLR4protein (immunohistochemical staining, Western Blotting), Fas(Western Blotting),TLR4mRNA(RT-PCR), and renal cell apoptosis (TUNEL)．(4)Statistical analysis: All statistical analysis of the experimental data were used CHISSstatistical software package, the results were expressed as mean±standard deviation (—X—±s).Differences are compared using single factor analysis of variance (ANOVA), correlationanalysis using Pearson correlation test, the P<0.05was considered statistically significance.Results:(1) General changes: Normal control group rats could move freely, the mentalstate is good, responsive, and normal diet; exhaustive exercise group rats were unable tomove, apathetic, indifferent response, lower diet. The state of Anisodamine group rats wasbetter than that in the same period of exhaustive exercise group rats.(2) The renal tissuestructure was no significant changes in CN and the expression of TLR4,Fas,TLR4mRNA,renal cell apoptosis were light. Renal tissue pathological changes were increasedprogressively from ES0h to ES24h (P<0.05). The expression of TLR4,Fas,TLR4mRNAand renal cell apoptosis were significantly increased in ES compared with CN(P<0.05). Theexpression in AD was significantly lower than the same period(P<0.05).(3)Respectively,TLR4protein had significantly positive correlation with the value of Fasprotein and renal tubular cell apoptosis (r=0.8102,P<0.05；r=0.848,P<0.05).Conclusions:(1) Overtraining can induce renal tubular cells apoptosis through activatingFADD signal pathway by impairing TLR4.(2) Pretreatment with Anisodamine inhibited theup-regulation of TLR4, Fas and cell apoptosis in renal tublar cells induced by exhaustiveswimming, which suggest that it play a significant protective effect on rats of OTIAKI.