| Objectives:Block the contact of the γ chain family of cytokines and JAK-STAT signaltransduction pathway by the CP-690550, to explore its impact on the γ chain familyof cytokines in the immediate immune, and ischemia-reperfusion myocardial clearprotective effect.Method:We adopt the heart transplantation of rat to construct the model of injuryinduced by myocardial ischemia/reperfusion: Take out the donor heart and match theaortic and pulmonary artery of the donor heart with the postcava and aortaventralis,respectively. After the surgery, if the donor heart beats again and the donor rat issurvive, which means the model construction is successful.The experiment is divided into5groups:6control groups,6model groups,6CP-690550high dose groups,6CP-690550medium dose groups and6CP-690550low dose groups. As the heart transplantation model of rat is successful constructed,we lavage each group with the CP-690550of different concentration and respectiveamount saline for model group and control group. After12hours, each group is doseagain. The rat is put to death after24hours, taking blood from abdominal vein andgathering the donor heart. Then we keep the serum and test myocardial tissue andorganization homogenate according to related index.Detection method and observation indicators:1. The variations of IL-2, IL-4, IL-7, IL-15in serum are test through EnzymeLinked ImmunoSorbent Assay (ELISA) and related biochemical method is used todetect the content of CK, LDH,SOD, MDA, in myocardial tissue.2. The sappanwood fine-Iraq red dyes is used to observe the variations ofmyocardial morphology.3. The expression of Bcl-2and Bax protein in myocardial tissue is detected by using SABC method.4. The apoptosis of cardiocytes is detected by using Terminal-deoxynucleoitidylTransferase Mediated Nick End Labeling (TUNEL).Results:1. Compared to normal group, the detection results of model group indicate that theexpression of IL-2, IL-7, IL-15(P<0.001) is improve, but which is reduce for IL-4（P<0.001）. For drug group, the opposite trend of γ-chain family cytokine is appear,the expressions of IL-2, IL-7, IL-15are reduce (P<0.05) but which is improve forIL-4(P<0.05).2. Compared to normal group, the detection results for model group indicate that:the activity of SOD（P<0.001）、CK（P<0.001）、LDH（P<0.05） are reduce, thecontent of MDA is increase (P<0.01). For drug group, the trend is opposite, SOD、CK、LDH (P<0.05) is improve, which is reduce for MDA（P<0.05）.3. For model group, myocardial cell get swell and deformation and a part ofmyocardial fibers horizontal grain is not clear or disappear. Compared to modelgroup, the degree of myocardial cell being degeneration of drug group decrease.4. For model group, the expression of Bax is increases. Bcl-2is not been expressedand myocardial cell appear a large apoptosis. Compared to model group, theexpression of Bax and Bcl-2are decrease and increase, respectively. Furthermore,the degree of myocardial cell apoptosis is alleviates.Conclusions:1. When ischemia-reperfusion injury happens, the content of promote inflammationfactors IL-2, IL-7and IL-15are improve greatly, and the content ofanti-inflammatory factor IL-4are decrease greatly. The γ-chain family cytokineclosely related to the injury. By block the interfere from JAK3-STAT signaltransduction pathways to the expression of the γ-chain family cytokine,ischemia-reperfusion injury myocardial is been protected.2. CP-690550can alleviate the myocardial damage for heart transplant ischemia-reperfusion model, the mechanism is:1) intervene γ-chain family cytokine: decrease promote inflammation factors and increase anti-inflammatory factor.2) resistance tooxygen free radical damage: increase the activity of myocardial SOD, CK and LDHand decrease the content of MDA.3) increase anti-apoptotic protein BCL-2onmitochondria apoptosis pathways and decrease the expression of promote apoptoticprotein Bax by blocking JAK3-STAT signal transduction pathways. |
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