| Viral myocarditis is a mostly common disease which happened on pediatriccardiovascular，it is the onset of an upward trend in recent years and has become oneof the main reasons of teenagers unexplained sudden death, moreover, chronic VMCcan also lead to the occurrence of dilated cardiomyopathy, serious harm to life andhealth in children. By far, more than20types of viruses have been now known tocause myocarditis, specific affinity coxsackie B3virus (Coxsackievirus B3, CVB3)most common on myocardial infection in the crowd. By far VMC pathogenesis hasnot been fully elucidated.12-lipoxygenase (12-LO) is a class of multifunctionalenzyme prosthetic group to the non-heme iron protein, which belonging to a lipidenzyme family, has been widely distributed in the various parts of the body and has avariety of biological functions.12-LO is an important signaling molecule, canpromote cell proliferation through oxidative stress response and inflammation, andparticipate in a variety of diseases pathological processes, and plays an important rolein the cardiovascular,involved in high blood pressure,atherosclerosis atheroscleroticdisease pathological process. The purpose of this study is to investigate the impact of12-LO and12-LO inhibitor skullcap against the incidence of viral myocarditis inmice, the feasibility of providing theoretical and experimental basis for the12-LOinhibitor baicalein treatment of viral myocarditis,12-lipoxygenase inhibitorbaicalein inhibition of12-LO activity is expected to treat the VMC.（1） The role of12-LO in the pathogenesis of viral myocarditisBALB/C mice by intraperitoneal injection of Coxsackie virus B3(CVB3)making the classic model of viral myocarditis, the establishment of a mouse model ofviral myocarditis. Take the4-week-old BALB/c male mice120, weight13~15g,were randomly divided into two groups, the control group (n=40) and viral model group (n=80), the virus model group mice were inoculated intraperitoneally0.1ml102TCID50CVB3virus to establish VMC model, while the control groupEagel’s virus intraperitoneal injection of the same amount of virus-free culturemedium (MEM). All mice conventional breeding. In four days after the virusinoculation,7days,14days,21days randomly portion mice, weighing wereobserved in mice changes in appearance,timely collect peripheral blood, centrifugedto obtain serum spare then dislocated killed, quickly take heart, dry filter paperweighed, and then fixed with formaldehyde made after histopathological check. Byreverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistrywas used to detect12-LO changes in the myocardium, fluorescent probe calciumimage analysis to detect intracellular free calcium (Ca2+) changes, the simultaneousdetermination of the myocardial pathology integral, serum creatine kinase MB mass(CK-MB mass), a Nitric Oxide (NO) and other changes, observed CVB3infectiontime12-LO and related indicators of changes in the pathogenesis of VMC, and CVB3infection myocardial pathologic score of myocardial tissue12-LO, and myocardialpathology scores and12-LO, VMC pathogenesis research and clinical treatment toprovide a new theoretical foundation and experimental basis. The results show that:the model mice, three days after the injection of the virus disease, mice, emergingreduced activity, Eat, hair stood on end, the color becomes dark, slow to react tostimuli, began to appear in the first four days after the death of seven days to14daysfor the death of the peak (15, accounting for65.2%of the deaths), gradually reducesymptoms Rule14d,17died throughout the experiment, the mortality rate was28.3%, control mice, no deaths occur, the survival rate of100%. Model mice ateach time point myocardial pathological changes and inflammatory cell infiltration,myocardial necrosis, the most serious of7days-14days Mingkesaqi table B3virusintraperitoneal injection can successfully build Balb/c mice myocarditis model.Semi-quantitative RT-PCR analysis Coxsackie B3virus infection after four days,7days,14days and21days in the myocardial tissue12-LO expression changes,wefound four days after infection, the expression of12-LO starts to rise, and thengradually increase to seven days, and reached the peak, four days after infection,7 days,14days and21days of12-LO expression than normal control group. ThatCVB3infection after14days of myocardial tissue expression of12-LO presents atime-dependent manner. Linear correlation analysis to determine the12-LO withcardiac pathology score, serum CK-MB mass has positive correlation, suggestingthat12-LO VMC may be involved in the occurrence and development. And of nitricoxide (NO), intracellular Ca2+carried correlation analysis, found that12-LO and ofNO, intracellular Ca2+was being related to sex, prompted12-LO may is NO andintracellular Ca2+the precipitating factor.(2)12-LO inhibitor baicalein treatment effect of viral myocarditis in mice240Balb/c male mice were randomly divided into five groups the normal miceblank control group (n=40), the animal model mice in the control group (n=50),high-dose treatment group (n=50), in the dose treatment group (n=50), low-dosetreatment group (n=50). The VMC model and specimen collection method with thefirst part of. CVB3infection in order to study the role of12-LO inhibitor baicalein inthe pathogenesis of VMC, two hours after intraperitoneal injection0.4mg/mL,0.2mg/mL,0.1mg/mL three doses, namely high, medium and low dose baicalein0.5ml. Myocardial tissue HE staining light microscope morphological changes inmice,using the semiquantitative score plus the pathological score, using RT-PCRdetection of myocardial12-LO changes,using fluorescent probe calcium imageanalysis to detect intracellular Ca2+changes,using colorimetric determination of theserum concentration of NO, observed12-LO,12-LO inhibitor baicalein and eachgroup of mice indicators of change. The results showed that: By comparison of the12-LO inhibitor baicalein treatment group and the mouse model and blank controlgroup, confirmed the decreased expression of12-LO12-LO inhibitor baicaleintreatment group at each time point, myocardial pathology score was reduced, serumCK-MB mass lower, at the same time,the content of NO, the intracellular Ca2+concentration also reduced; inhibiting effect in a dose dependent manner, theinhibitory effect is most obvious high-dose group, the low-dose group suppressioneffect weakest, and12-LO expression was positively correlated. Tips baicaleinreduce myocardial inflammation in mice by inhibiting the expression of12-LO, improve myocardial injury in mice. Baicalein inhibition of12-LO activity in thetreatment of viral myocarditis, and its possible mechanism of the inhibition of NOexpression, lower intracellular Ca2+concentration.Conclusion:Comprehensive on the results of this study suggest that: compared with normalcontrol group,12-Lo is significantly increased in the myocardial tissue of mice withviral myocarditis，the pathological score of12-LO expression levels in myocardialtissue of mice with viral myocarditis, CK-MB mass were positively correlated,suggesting that12-LO with viral myocarditis mice myocardial tissue damage processclosely related to the severity, which can be regarded as reflected factor in the degreeof the inflammatory and indicators of myocardial necrosis, indicating that the12-LOplays an important role in the development of viral myocarditis.12-LO expressionamount of iNOS and NO intracellular Ca2+was a positive correlation, suggesting that12-LO may be a precipitating factor NO and intracellular Ca2+.12-LO inhibitorbaicalein has some therapeutic effect in mice with viral myocarditis.Intervention therole of12-LO can reduce the viral myocarditis lesions.12-LO inhibitor baicalein maybe effective in the treatment of viral myocarditis. This study clarifies the role of12-LO in the pathogenesis of viral myocarditis, providing a potential avenue for theprevention of viral myocarditis and a new idea for other inflammatory diseaseresearch. |
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