The Mechanism Of Immune Inflammation Coupling Endoplasmic Reticulum Stress Involved In Heart Failure And The Intervention Role Of Immunoglobulin

Heart failer always accompanied with abnormal activation of immune inflammatorysystem marked as elevated levels of cytokines. And the cytokines in heart failure can startand aggravate heart failure through inhibiting myocardial contractility,promoting cardiacmyocyte hypertrophy,apoptosis and extracellular matrix metabolism.For cytokines increaseits own synthesis through a positive feedback mechanism,so sustained activation ofcytokine will lead to heart failure. TNF-alpha,IL-6play a key role in heart failuremechanisms,whether they can work as BNP for the diagnosis of heart failure,picardiacfunction evaluation and disease monitoring or prognosis,or become the target for thetreatment of heart failure is the focus of current research. In the treatment of heart failure,there is no significantly decrease incidence of heart failure and mortality although manyantagonistic measures was carried out basing on extensive research in theimmune-inflammatory pathogenic role and its mechanisms.Thus,finding a key target forheart failure of immunotherapy is nervous.Endoplasmic reticulum stress (ERS) can correct misfolded proteins,enhance cellularadaptation and ability to resist stress,but if the stress intensity is high or duration is toolong,the ERS will activate the corresponding cell signal transduction and leadto celldamage,and thus participate in immune and inflammatory response. ERS can induceNF-B activation,which mediated immune inflammation; can induce NF-B inhibitionfactor A20expression in turn to inhibit NF-B activity,decreased the expression ofproinflammatory cytokines and induce disability of the cells in the immune. Inflammatorycells caused ERS mediated by the ROS and ROS promote the inflammatory response.Therefore,the coupling of ERS and immune-inflammatory response in the specific cellsinvolved in the heart failure happen,development. However,the relationship betweenexcessive activation of immune inflammation and the ERS response in heart failure wasrarely reported.at home and abroad. Immunoglobulin (IVIG) is a series of protein which have immune substitution andimmunomodulatory activity,can mediate the proliferation and chemotaxis of monocytes-macrophages,lymphocytes and other immune cells,mediate the release of inflammatorymedium and NF-B transcriptional activity,play specific anti-inflammatory effects in theimmune and inflammatory disease. However,how IVIG regulate the response degree ofendoplasmic reticulum stress in myocardial injury,how control heart failure diseaseprogression has not been exactly explained.Our experiment detected the correlation between cytokine expression and cardiacfunction in experimental heart failure and their relationship with the ERS,analysis thecytokines'effection in the happen and development heart failure from the level of clinical,the general and molecularthe possible molecular mechanisms of the couplingimmune-inflammatory and ERS induced heart failure.Through observating theserelationships' change after the intervention of IVIG,revealing the immuno modulatoryeffects of IVIG in heart failure therapy and its possible mechanism to provide theexperimental basis for clinical application of IVIG.Method:The patients of heart failer was divided into tree groups based on cardiac function,NYHA class II group,NYHA III group and NYHA class IV group,taking heath personsas in the same period as healthy controls,detecting serum TNF-a,IL-6and BNP levels byELISA assayLigated anterior descending branch of the left coronary artery of female Wistar rats toestablish myocardial infarction heart failure animal model,at the same time taking shamoperation group as controls.Animals were randomly divided into model group and the IVIGgroup7weeks after surgery,then were given saline and IVIG via intraperitoneal injectionsindividually for5weeks. Detected the emodynamic parameters; morphology chang of thecardiac,observated the myocardial cell morphology and proliferation of collagen fibers viaHE and Masson staining; determinated TNF-alpha,IL-6mRNA expression levels byRT-PCR,detected myocardial tissue TNF-alpha,IL-6protein level of by immunestaining,detected cardiac myocyte apoptosis by TUNEL staining; Determinated thymusand spleen lymphocyte proliferation and secretion capacity of TNF-alpha,IL-6; usingwestern blot to detecte TNF-,IL-6and GRP78protein expression. Result:The serum levels of IL-6,TNF-and BNP increased dramatically campared withhealthy controls and had positive relationship with the degrees of heart fuction,thefurther analysis showed that the surum levels of IL-6,TNF-of the patients with heartfailar had positive relationship with the surum levels of BNP. Compared with the shamoperation group,LVSP and±dp/dtmaxof model group rats were significantly decreasedand LVEDP significantly increased,heart hypertrophy index (HW/BW) and leftventricular hypertrophy index (LVHW/BW) was significantly higher; LVSP and±dp/dtmaofmodel group rats and the HW/BW and LVHW/BW significantly reduced. after IVIGintervention. Morphological test results showed that: heart volume of the model ratsincreased,the geometry of hearts changes,the ventricular chamber expansed,aneurysmformation. HE staining showed myocardial cells of models arranged in disorder,haddifferent degrees of degeneration and hypertrophy. The cardiac volume of IVIG group ratsincreased less significantly compared with the model group,maintained the normalgeometry basically; less aneurysm formation and ventricular chamber dilatation comparedwith the model group.HE staining showed that cardiac cell swelling of IVIG-treated group is significantlyreduced compared with the model group. Masson staining shows the myocardial cellshypertrophy in the model group,cell number reduce and disorganize,a large number ofcan be seen in the cell gap,green collagen fibers significantly decrease in IVIG group..Serum TNF-alpha,IL-6,BNP of Model group significantly increased compared withthe sham group and significantly decreased after the intervention of IVIG,and serumTNF-alpha,IL-6levels showed a positive correlation with serum BNP levels. Theexpression of TNF-alpha,IL-6mRNA and protein in myocardial tissue of model groupincreased compared with the sham operation group and significantly decreased after theintervention of IVIG.. TUNEL staining found that more cardiomyocyte apoptosis in modelgroup than sham operation group and using IVIG could inhibite cardiomyocyteapoptosis.The lymphocyte transformation test results showed that lymphatic stimulationindex of model group was significantly higher than sham operation group. TNF-,IL-6levels of lymphocyte culture supernatant increased significantly compared with the shamoperation group. Lymphocyte proliferation and TNF-,IL-6level in cell culture medium of IVIG group were significantly lower compared with model group. The expression ofGRP78,the marker protein of ERS response,increased in the myocardial tissue of modelgroup and the application of IVIG can significantly decreased the level of GRP78proteinexpression.Conclusion:1. The immune system was activated in heart failure,the performance of theinflammatory cytokines TNF-αlpha,IL-6in serum,protein and gene expression levelswere significantly higher,had positively correlated with the grade of cardiac functional andserum BNP levels2. Immunoglobulin administration can reduce TNF-alpha,IL-6expression levels,improve heart function of rats with heart failure,suggesting that IVIG maybe improve thecardiac function by suppressing abnormal activation of the the immune system.3. Cardiac hypertrophy,myocardial fibrosis and cardiac myocyte apoptosis wereoccered in rats with heart failure. Immunoglobulin can stabilize the normal structure of theheart,inhibit proliferation of myocardial fibers and myocardial apoptosis,and protectstructure and function of the heart.4. The proliferation and differentiation of T and B lymphocyte were enhanced,and itssubsets is activated in rats with heart failure,the expression of GRP78that is landmarkprotein of ERS was increased,suggesting that immune inflammation is associated withERS coupling to participate in the development of heart failure.5. IVIG can inhibit the elevated GRP78and TNF-alpha,IL-6protein,suggesting thatthe IVIG to improve the possible mechanisms of cardiac function is inhibitted ERS in orderto suppress the activation of immune system.