The Inflammatory Injury Study Of Toll-like Receptor Signal Pathway Of Fluoride On Cardiac In Rat

[Objective]With the further study on the fluoride,there are many researches on the damage of fluoride to the soft tissue.However,fewer researches is known about the dynamic effects of fluoride exposure on the body and the molecular mechanisms underlying cardiac inflammation.In this research,we will study the toxic effects of different doses of NaF on the effects of rat's heart.Using high-throughput sequencing technologies to explore the molecular mechanisms of cardiac immune injury in rat exposed to fluoride,and provide the basis theoretical for further consummating cardiac immune injury mechanism of fluoride.[Methods]Using fluoride exposure method to establish the experimental model in rats,rats with similar body weight were selected and randomly allotted to 3 groups:control group,low fluorine group(30mg/L NaF)high fluoride group(90mg/L NaF);during the period of infection,observed and recorded the changes of diet and body weight.After 6 months take's heart tissue of rats,using light microscope and transmission electron microscope to observe morphology,measuring metabolic enzyme activity in myocardial tissue,using RT-qPCR method to evaluate the expression of the cardiac inflammatory pathways related to mRNA(CD14,MR,Fcry,IRAK1/2,CNK,CARD9,IL-1,IL-6,IL-10,TGF-beta,V-ATPase).using Western Blotting to detect the expression of IL-1,IL-6 and IL-10 protein in rat heart,and the correlation analysis was carried out to study the cardiac toxicity of fluoride[Results]1.Compared with the control group(Control),there was no significant difference in body weight between the experimental groups,while the weight of the rats in the experimental group decreased with time grows(p<0.05).2.Rat myocardial tissue by HE staining showed that the control group myocardial cell myofilament and muscle fiber arranged orderly,clear stripes,cell morphology,the nucleus is clear,uniform staining,cell gap junction closely;low fluorine group myocardial cell myofilament arranged loose,but the overall change is not obvious;the high fluoride group Center muscle muscle fibers arranged disorderly,and even the emergence of large transverse fracture,cannot significantly observed,cell gap increases,the gap with a large number of bleeding points,nuclear condensation,cell rupture.3.The myocardial tissue TEM results show that:in the control group,the myocardial tissue structural integrity,myocardial cells arranged orderly,muscle fiber integrity,Z line,M line is clearly visible,the number and morphology of mitochondria were normal,intercalated disc connected complete;Effects of low concentration on the structure of myocardial cells in rat left ventricular muscle fiber fracture is not obvious,appear very little muscle fiber fracture;the muscle fiber appeared serious faulted,cardiac cell was disorder and swelling,the Z line,M line was broken obviously,the structure of most mitochondrial was broken and appeared the vacuoles,intercalated disconnection completely interrupted in the high group.4.Metabolic enzyme activity in myocardial tissue showed that:compared with control group,creatine kinase(CK),aspartate aminotransferase(GOT)and creatinine(CRE)were increased in the high fluoride group.5.Using RT-PCR to detect the expression of immune related of genes CD14,MR,Fcry,IRAK1/2,CNK,CARD9,IL-1,IL-6,IL-10,TGF-? V-ATPase in rat heart.The results showed that,compared with control group the expression of CD14,MR,Fcry,IRK1/2,CNK,CARD9,IL-1,IL-6,IL-10,TGF-? mRNA were decreased in both treatment(p<0.001,p<0.01,p<0.05),while the expression of V-ATPase increased in high fluoride group(p<0.01,p<0.05).6.Western-blotting was used to analyze the expression of IL-1,IL-6,and IL-10 protein in the myocardial tissue of rats.The results showed that IL-1,IL-6 and IL-10 proteinin each treatment group were significantly lower than those in the control group(p<0.001,p<0.01,p<0.05)[Conclusion]In the expression,fluoride exposure injury of rat heart tissue for structural damage of myocardium,the macro level is confirmed by transmission electron microscopy and cardiac enzyme assay,at the micro level on the molecular mechanism of injury research shows that fluoride effects on the heart may be through the impact of changes of immune related genes in the inflammatory pathway to achieve.