| BackgroundSepsis is due to microbial or other pathogens invade the human body induced by excessive intense systemic inflammatory response, physiological and pathological processes and a set of clinical manifestations, it may develop to severe sepsis and septic shock. There are more than 18 million global cases of severe sepsis each year. The worry is that the incidence is 0.3%, the rate will increase by 1.5%, to 2020, it will increase to 1 million cases each year in the United States. Now in the United States, more than 75 million people suffering from sepsis each year, of which more than half patients with sepsis stay in ICU about 15.7 days.During the development of sepsis, as it is difficult to control the body’s inflammatory response, multiple organ lead to acute injury, and then develop severe sepsis, septic shock and MODS. The kidney is one of the most commonly attacked organ. Acute kidney injury (AKI) is one of the most common complications of sepsis. Kidney is a special vascular organ in the body, which will lead to the disturbance of machine body environment, and then induce or aggravate the injury of other organs of the body, leading to MODS at last. Although AKI has such a high incidence and mortality rate in patients with sepsis, the study of AKI induced by sepsis is relatively less. Much studies have shown that if renal function of patients with sepsis in mild impairment,it can lead to the increase the fatality rate; if we can intervent at the moment that the glomerular filtration rate (GFR) is still in normal or just slightly decreased the treatment can significantly reduce the mortality of septic patients. Therefore, it is urgent to study the mechanism of pathological and physiological changes of renal injury in patients with sepsis and to find a new effective method for the treatment of AKI.The activation of NF-kappa B can enhance the transcription of TNF, IL-1 and IL-2 genes, and can increase the production and release of TNF- alpha, IL-1 beta and IL 2, and then activate the NF-nuclear factor kappa B. It can also increase the levels of IL-6 and IL-8 of these inflammatory cytokines, which further amplify the signal that causes the initial inflammation and aggravate the injury of the body’s microcirculation until DIC or death. This situation is more common in Sepsis Syndrome and acute respiratory distress syndrome (ARDS). But this situation does not occur without limitation in the body of the disease process, because NF-kappa B has a negative feed back regulation.Resveratrol (Res) is a kind of non-flavonoid phenols. It is a natural resveratrol in many plants, it is a type of plant material to resist bacteria poisoning caused by intrusion. Resveratrol has a strong antioxidant capacity, and it is a non synthetic plant antioxidants, studies have shown that it can inhibit platelet aggregation, vasodilate blood vessels, reduce blood viscosity, also can inhibit the tumor, is usually used for the treatment of coronary heart disease, hyperlipidemia and ischemic heart disease.The study also found that resveratrol is similar to estrogen action. Therefore, it is to try to treat for breast cancer. In the 90’s of last century, China has begun to study resveratrol, some people found that it had a certain therapeutic effect on the heart and tumor, but also can play a role in weight control, improve microcirculation and so on. A study found that resveratrol can improve the renal microcirculation after CLP, perhaps by reducing the free radicals of reactive nitrogen, thereby protecting the renal tubular epithelial cells.Serum creatinine, urea nitrogen and urine volume are the classic standard of AKI. Large quantities of data have confirmed that early AKI biomarkers include neutrophil gelatinase associated lipid carrier protein (NGAL), kidney injury molecule-1 (KIM-1), L type fatty acid binding protein (L-FABP), alpha glutathione s transferase (alpha GST) and interleukin 18 (IL-18), these markers raised 24-48 hours earlier than serum creatinine. Therefore, the detection of these markers is helpful to the judgment for subclinical AKI, and it has important significance for the prognosis of the patients. Recent studies have indicated that IL-18, NGAL, and KIM-1 are predictive of relatively strong signaling molecules in sepsis AKI. For the majority of renal injury, the increase of concentration and proportion of biomarkers in urinary are significantly higher than plasma, for example, the increase of urinary cystatin C (cystatin C) and albumin indicate that the dysfunction of proximal tubular reabsorption.Over the past two years, people gradually began to pay attention to the cycle of miRNA as a biomarker of AKI. Usually only a biological mark level increases more, and a variety of other biological marker elevated levels of consistency, and accompanied by the decreasion of glomerular filtration rate and urine, people can think that the biomarkers and severity of AKI and clinical prognosis correlation is stronger. But only a single, with sufficient sensitivity and specificity of the markers did not reflect the whole process of AKI pathophysiology and disease alters.NF- nuclear factor kappa B in the kidney injury related diseases are widely concerned.The researchers found that the expression of NF-nuclear factor kappa B will rise when the kidney is injury, and then progress to a series of inflammatory reaction.Resveratrol possesses a variety of bioactivities,including anti-inflammatory, antioxidation and improving microcirculation effect. In recent years, animal experimental study on the treatment of sepsis were obtained more positive results.It is that resveratrol can effectively improve the sepsis induced injury, including lung injury and brain injury. Studies have also found that resveratrol can improve many types of kidney injury, including diabetic kidney injury, ischemia reperfusion injury, drug related injury etc.We try to think that NF kappa B will have some changes after sepsis acute kidney injury, but specifically how to change, which factor will be regulated to, it will change or not after the intervention of resveratrol, it is the main purpose and direction of our research.ObjectiveTo investigate the pathogenesis of septic AKI, seeking ways to protect sepsis organ suffered from acute injury. This study intends to making sepsis induced AKI model through cecal ligation and puncture,in rats, It imitated the clinical course of infection of abdominal sepsis, causing multiple bacteria infection, and eventually leading to sepsis. To preliminary study on protective effect of resveratrol on AKI in septic rats and possible mechanisms of action, provide a more theoretical and experimental basis for clinical treatment of sepsis in application of resveratrol.Methods1.Twenty SD male rats were feed and divided into Sham group and CLP group randomly, two groups were established by using the CLP sepsis model, Oh,6h,12h, 24h was the time we chose to take blood, ELISA was used to detect changes in renal and inflammatory factors, to clarify 12h was the successful point time that we established sepsis kidney injury model successfully.2.Fifty adult male mice were randomly divided into five groups after normally bred.They were divided into Sham, Sham+RES, CLP, CLP+RES (3mg/kg) and CLP+RES 10mg/kg) five groups randomly. Ten male SD rats were assigned to a group,the weight of rat was about 170-220 gram. Making pathological section to observe renal histological changes.Using ELISA method to measure serum renal damage index、changes of inflammatory factors and the animal survival rate.3.Using glomerular mesangial cells to make model in vitro.Using RES to invervent the glomerular mesangial cells,we test the NF-κBP65 and SIRT1 protein expression by western bloting method,so as to explore the mechanism that the RES could regulate the NF-κBP65.4. SPSS 17.0 statistical software was used for statistical analysis Statistical analysis:results are presented as mean±standard deviation (x±s) with multiple replications in each experiment. Analysis of variance, one-way ANOVAor Welch test was used. The Least-significant Difference (LSD) or Dunnett’s T3 test was used to evaluate differences between means, with a P-value less than 0.05 being considered significant.Results1.The difference of the serum Scr、BUN and other indicators in the Sham group at different time point was not statistically significant (P> 0.05); but in the CLP group, the serum Scr、BUN、ScysC、NGAL、KIM-1、TNF-α、IL-1β、IL-6、 IL-10、had significantly improved over time. Compared with 0 hour,the difference was statistically significant (P<0.05).2.The different of concentration of the serum Scr、BUN、ScysC、NGAL、 KIM-1、TNF-α、IL-1β、IL-6、IL-10、the renal blood flow、GFR and the renal injure index in Sham group and Sham+ RES group were not statistically significant (P>0.05); but in the CLP+RES group,the serum NGAL、KIM-1 concentration decreased after the intervention of resveratrol compared with the CLP group. With the increase of RES concentration, the effect was more obvious, RES intervention group and CLP group had a statistically significant difference (P<0.05), the serum Scr、BUN、Scys C、TNF-α、IL-1β、IL-6、the renal injure index were also simmilar to the above, while in renal blood flow、GFR and IL-10, the situations were also opposite to the above, the CLP+ RES group with different concentrations could be clearly seen in renal blood flow、GFR and IL-10 after the intervention of resveratrol and the CLP group improved significantly, and with concentrations of RES increasing, it improved significantly, there was a significant statistical difference in the RES intervention group and the CLP group(P<0.05).3.The kidney tissue biopsy showed that the pathological changes of glomerulus、renal interstitial and renal tubules in rats of the CLP+RES groups alleviated obveriously.The infiltration of the inflammatory cells of kidney tissue alleviated than the CLP group in corresponding time.The surrival time and surrival rate in the CLP+RES groups were higher than the CLP group.It also proved that resveratrol had a significant effect on controlling and reversaling AKI.4.Between the CLP group and the Sham group after operation, NF-κB P65 protein levels were significantly upregulated, and the difference was statistically significant (P<0.05); and in the CLP group and the CLP+RES group, it could be clearly seen that the NF-κBP65 protein levels significantly decreased compared with the CLP group, there was significant statistical difference (P<0.05) in the RES intervention group and the CLP group. The SIRT1 was opposite.5.The SIRT1 protein levels of LPS group was significantly down-regulated compared with the Control group, and the difference was statistically significant (P <0.05); while in the LPS+RES group,it could be clearly seen that SIRT1 protein levels after the intervention of resveratrol significantly increased compared with the CLP group, there was a significant statistical difference between the RES intervention group and the LPS group(P<0.05); and after transfection of cells of SIRT1 RNAi to block the SIRT1, you could see the SIRT1 protein levels decreased significantly.6.The NF-κBP65 deacetylation of protein expression levels were significantly upregulated during the detection of protein acetylation between the Control group and the LPS group, and the difference was statistically significant (P<0.05); while in the LPS+RES group,it could be clearly seen that the expression of NF-κBP65 deacetylation protein in the LPS group significantly decreased after intervention of Resveratrol, there were significant statistical difference between the(P<0.05) RES intervention group and the LPS group; and after transfection of cells of SIRT1 RNAi to block the SIRT1, you could see the NF-κBP65 deacetylation of protein expression levels significantly increased.Conclusion1.Through establishing cecal ligation and puncture (CLP) model of sepsis, it can replicate the animal pathology of acute kidney injury.2.Resveratrol can alleviate acute kidney injury, significantly reduce the mortality in septic rats,one of the mechanisms is to reduce the kidney tissue inflammation.3.Resveratrol can cause NF-κBP65 to deacetylation through raising SIRT1 significantly, and reduce production of inflammatory cytokines and inhibit inflammation injury to the kidneys. |
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