Clinical Observation Of Parecoxib On Stress Response And Inflammatory Response Of Thoracoscopic Surgery And Spectroscopic Study On The Interaction Mechanism Between Parecoxib And P38MAPK

BackgroundAt present,although the mainstream thoracoscopic radical resection of lung cancer has the advantages of small incision,less injury,subtle operation,less bleeding and so on.However,surgical injury of lung tissue and lymph node dissection and other factors,patients' stress response and inflammatory response are still large;postoperative pain affects the normal cough and expectoration of patients,and the demand for postoperative analgesia is still high.The quality of postoperative rehabilitation is the result of the interaction of many factors:the mode of operation,the time of operation,the degree of stress and inflammatory reaction,the quality of intraoperative anesthesia,the quality of postoperative analgesia,postoperative complications and so on.As early as 2001,Mr.Kehlet,a Danish surgeon,put forward the concept of rapid rehabilitation ERAS(Enhance Recovery After Surgery),which aims to reduce the average hospital stay and improve the quality of postoperative recovery.In the process of ERAS,inhibition of stress response and effective postoperative analgesia are the key links.After thoracic surgery,patients are afraid of pain,dare not take a deep breath,cough and expectoration are limited,resulting in retention of respiratory secretions,blockage of small airways,easy to cause atelectasis,falling pneumonia,and further aggravating inflammatory reaction.Prolonging the length of stay increases the cost of treatment and even threatens the lives of patients.Effective inhibition of perioperative stress and inflammatory response and perfect postoperative analgesia are beneficial to improve the quality of life of patients and reduce the incidence of long-term complications.This experiment was designed to speed up the postoperative rehabilitation of patients,using general anesthesia combined with paravertebral nerve block and intravenous application of parecoxib to speed up the postoperative recovery of patients and provide clinical evidence for the rapid rehabilitation of thoracoscopic patients in the future.Non-steroidal drugs have a good anti-inflammatory and analgesic effect,the traditional non-steroidal anti-inflammatory is diminishing in drugs clinical application due to nausea,vomiting,stomach bleeding and other side effects.Parecoxib is the second generation of COX-2 inhibitor.After intravenous injection,it can be hydrolyzed by liver carboxylesterase to valdecoxib,a highly selective COX-2 inhibitor.It specifically inhibits the anti-inflammatory and analgesic effect of COX-2 by blocking the synthesis of prostaglandins from arachidonic acid.Because of the high selectivity of parecoxib to COX-2,the safety of parecoxib in gastrointestinal tract was higher than that of traditional NSAIDs,and had little effect on platelets and no effect on bleeding time.Preemptive analgesia with parecoxib can avoid central sensitization and pain upregulation caused by intraoperative nociceptive stimulation,reduce perioperative stress and inflammatory response,and prevent and relieve postoperative pain.The results showed that phosphorylated-p38MAPK,was positively correlated with the expression of COX-2 protein.The inhibition of p38MAPK phosphorylation can significantly inhibit the expression of COX-2 protein.lt has been shown that there is a positive correlation between p-p38MAPK and COX-2 protein expression in body stimulated by p38MAPK,and the inhibition of p38MAPK phosphorylation can lead to a significant inhibition of COX-2 protein expression.A variety of pain-related mediators or receptors such as COX-2,interleukin-6,tumor necrosis factor-a are related to the activation of p38MAPK in spinal cord microglia.As far as p38MAPK is concerned,COX-2 is a protein with increased expression due to p38MAPK activation.p38MAPK plays a key role in a large number of cellular responses,such as stress and inflammation,and is closely related to cell development,growth and differentiation.At present,the relationship between the inhibition of stress response and inflammatory response by parecoxib and p38MAPK is not clear.In this study,we studied the interaction between parecoxib and p38MAPK under simulated physiological conditions by spectroscopy,and studied the effect of parecoxib on the structure of p38MAPK protein by circular dichroism spectroscopy and three-dimensional fluorescence.It is helpful to understand the action mechanism of parecoxib in body,lays a theoretical foundation for further study of its action mechanism,and has a certain guiding significance for the rational use of drugs and the design of drug derivatives.Part 1 Clinical observation for parecoxib on stress response and inflammatory response of thoracoscopic surgeryObiectonsThoracic postoperative pain affects patients with sputum or deep breathing,hinder postoperative rehabilitation,stress response and inflammatory response is also an important aspect of the recovery of thoracic surgery.A large number of perioperative proinflammatory cytokines production will lead to excessive stress,destruction of the balance between cytokines,excessive stress response will weaken the body's physiological reserves,thereby increasing the inflammatory response,stimulate systemic inflammatory response syndrome(SIRS).Postoperative pain mechanism is complex,inflammatory pain is also an aspect of postoperative pain.In clinical practice,multi-mode analgesia is advocated,and the combined use of drugs or analgesic techniques with different mechanisms of action has less complications.Parecoxib is the second generation of COX-2 inhibitor,by selectively inhibiting COX-2 prevent PG and thromboxane A2(TXA2),and then play an antipyretic.anti-inflammatory and analgesic effect.In this study,patients undergoing thoracoscopic surgery were observed to observe the effect of intravenous injection of parecoxib on stress response and inflammatory response and postoperative analgesia in thoracoscopic surgery,so as to provide basis for postoperative analgesia under the guidance of ERAS.MethodsIn this study,40 patients who underwent elective thoracoscopic lobectomy from March 2016 to March 2017were selected with the approval of the Medical Ethics Committee.The ASA grade was grade ? to grade ?,and the informed consent was signed before operation.All patients underwent general anesthesia combined with ultrasound-guided paravertebral nerve block.All patients underwent the perioperative management model under the concept of ERAS,two hours before the ban on drinking,not indwelling the urine catheter,without preoperative medicine,the use of standardized preoperative preparation and anesthesia process.General anesthesia before the ultrasound-guided paravertebral nerve block,injecting 0.5%ropivacaine 20ml,10min after blocking the anesthesia plane,requiring anesthesia plane in the T4-T8.Patients were randomly divided into control group(group C,n=20)and Parecoxib group(group P,n=20).Parecoxib group were injected with parecoxib 40mg(saline diluted to 4ml)on preoperative(T0),24h after operation(T1)while control group injected with placebo(0.9%NaCl solution 4ml).The arterial blood was collected 8ml before operation(TO),24h after operation(Tl)and 48 hours after operation(T2).The cells were centrifuged at 3500r/min for 15 minutes at 4 C.Take the upper plasma at-80 ? low temperature refrigerator.The levels of ET,TXA2.IL-6 and TNF-a in arterial blood were measured by volume sampling.The total number of opioid drugs,infusion volume,blood loss.operation time,VAS score at 24h and 48 hours,and postoperative hospital stay were recorded.Results1.the general situation of patientsA total of 40 patients met the criteria for inclusion in the study.There was no significant difference in age,sex,height,weight,fentanyl dosage,operation time.blood loss and infusion volume between the two groups(P>0.05).2.postoperative VAS score and hospital stayThe postoperative VAS score in the paroxib group was lower than that in the control group at Tl(P=0.004),and the postoperative VAS score in the paroxib group was lower than that in the control group at T2(P=0.004).Compared with the control group,the postoperative hospital stay in the parecoxib group was lower than that in the control group(P=0.007).3.stress response and inflammatory response indicatorsThere was no significant difference in the levels of plasma ET,TXA2,IL-6 and TNF-? between the two groups before operation(P>0.05).The levels of plasma ET,TXA2,IL-6 and TNF-? in the two groups at T1 were higher than those before operation(P<0.05),and the levels of plasma ET,TXA2,IL-6 and TNF-? at T2 in the two groups were higher than those before operation(P<0.05).Compared with the control group,the levels of plasma ET,TXA2,IL-6 and TNF-? in parecoxib group were significantly lower than those in the control group at T1(P<0.05),but there was no significant difference in the levels of ET,TXA2,IL-6 and TNF-? between the two groups at T2(P>0.05).ConclusionsIntravenous injection of parecoxib can improve the analgesic effect of thoracoscopic surgery,reduce stress response and inflammatory response,shorten the hospitalization time of patients,and accelerate the postoperative recovery of patients,which meets the requirements of multi-mode analgesia and rapid rehabilitation under the concept of ERAS.Part 2 Spectroscopic study on the interaction mechanism between parecoxib and p38MAPKObjectonsThe p38MAPK affect a variety of intracellular responses,with well-recognized roles in inflammation,cell-cycle regulation,cell death,development,differentiation,senescence and tumorigenesis.MAKP(mitogen-activated protein kinase)is a widely expressed serine/tyrosine kinase,which plays an important role in a variety of signal transduction pathways in mammalian cells.In recent years,it has been found that p38 signaling pathway is an important branch of MAPK pathway.Under the stimulation of pathological factors,nerve cells can activate p38MAKP and then activate MAKP signaling pathway to secrete inflammatory cytokines such as IL-1?(interleukin-1?)and TNF-?(tumornecrosisfactor-a).Causing nervous system damage.Studies have shown that the expression of COX-2 protein is positively correlated with p-p38MAPK,the activator of p38MAPK,and the inhibition of p38MAPK phosphorylation can significantly inhibit the expression of COX-2 protein.A variety of pain-related mediators or receptors are associated with the activation of p38MAPK in spinal cord microglia,such as TNF-?,IL-1?,NK-1(Neurokinin-1)receptor and COX-2.In addition,p38MAPK activation can further increase prostaglandin synthesis and participate in central sensitization by activating COX-2.Parecoxib decreased the stress response and inflammatory response during perioperative period.It is speculated that the mechanism of reducing the expression of TNF-a,IL-1 and other proteins may not only directly inhibit cyclooxygenase,but also play a role by inhibiting the pathway implicated by p38MAPK.In this study,we used spectroscopy-based methods to study the interaction between parecoxib and p38MAPK under simulated physiological conditions.Circular dichroism spectroscopy and three-dimensional fluorescence techniques were used to study the effect of parecoxib on the structure of p38MAPK protein.At the same time,the binding site number,binding constant and thermodynamic constant of the interaction between parecoxib and p38MAPK were calculated.MethodsThe interaction between parecoxib and p38MAPK was studied by transmission electron microscope,UV-vis absorption spectrum,steady-state/time-resolved fluorescence spectrum,circular dichroism spectrum,energy transfer and so on.ResultsUV-vis absorption,circular dichroism and three-dimensional fluorescence can reflect that the spatial structure and microenvironment of p38MAPK are changed due to the combination of parecoxib,resulting in the gradual loosening of the overall skeleton structure of p38MAPK.Some hydrophobic aromatic cyclic amino acids and polypeptide chains were exposed to the solution.According to the experimental results of fluorescence spectrum and transmission electron microscope,parecoxib can lead to the agglomeration of p38MAPK,and they can combine with p38MAPK at l:l to form parecoxib-p38MAPK complex through the action of non-covalent bond force-electrostatic force.Parecoxib has a certain effect on the fluorescence quenching of p38MAPK,and the fluorescence quenching mechanism is classified as static quenching.The affinity constant KA between parecoxib and p38MAPK at 25 degrees Celsius was calculated by Stern-Volmer equation to be 8.13×104/M.From the Van‘t Hoff equation,? G,A H and ? S are calculated to be-2.80x104J·mol-1,-1.62 ×105J·mol-1·K-1 and-450J·mol-1,respectively.This process is mainly a spontaneous molecular reaction driven by entropy.The binding distance between molecules is 3.95nm.ConclusionsIn this study,it was found that parecoxib caused changes in the spatial conformation and microenvironnent of p38MAPK.p38MAPK has a relatively loose skeleton structure,while the internal hydrophobic polypeptide chain and aromatic cyclic amino acids are constantly exposed.The analysis of all the theoretical data and experimental results show that parecoxib has a significant effect on the structure of P38MAPK,and provides the basic data for the triggering of p38MAPK pathway.