| OBJECTIVE: To study the mechanism of paeoniflorin inhibiting myocardial dysfunction via A1-adenosine receptor in septic mice.METHODS: 1. Health SPF male Kunming mice were used to reproduce experimental septic model with cecal ligation and puncture(C LP). And keeping the weight of these mice between from 30 g to 35 g. In preliminary experiments, we confirmed that dosage of 20 mg/kg and 1 mg/kg were the proper dose of paeoniflorin and DPCPX. These mice were randomly divided into 8 groups which was at least 5 mice each of groups. Groups were as follows: A. CLP+ DMSO+ H2O;B.CLP+ DMSO + Pae;C.CLP+ DPCPX+ H2O;D.CLP+ DPCPX + Pae;E. Sham+ DMSO + H2O;F. Sham+ DMSO + Pae;G. Sham+ DPCPX + H2O;H. Sham + DPCPX + Pae. 2. The mice would be sacrificed 12 hours later after the operation. Cardiac tissues from mice were immobilizated, fixed, embedded, H&E strained pathological sections and the morphological change was observed by florescent microscope. 3. To observe the effect of of various of cytokines in septic mice blood serum and cardiac tissue by paeoniflorin on the A1-adenosine receptor, 6 cytokines were measured with Luminex x MAP technique after the blood serum and cardiac sample were collected 6 hours later after the operation. The cytokines include interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), interleukin-12(IL-12), interferon-γ(IFN-γ) and interleukin-10(IL-10). 4. To study the mechanism of inhibiting septic and cardiac dysfunctional mice myocardial cell apoptosis by paeoniflorin on the A1-adenosine receptor. Western blot method were used to evaluate the myocardial phosphorylated and unphosphorylated proteins levels of P38, ERK, JNK and the expression level of Bcl-2.RESULTS: 1. Under the microscope, a lot of histological lesion of myocardial cell in CLP model groups was observed, such as edema, a disorganization of myocytes. Compare with other C LP groups, paeoniflorin theropy group could obviously relieve the pathological changes of cardiac tissue. DPCPX groups are the opposite. All of Sham groups appear a normal morphology. 2. For blood serum, all of the level of 6 cytokines in CLP groups are evidently higher than Sham groups(P<0.05). Except for IL-10, the CLP+ DMSO+ H2 O group’s values are remarkably higher than C LP+ DMSO + Pae group(P<0.05), but lower than CLP+ DPCPX+ H2 O group(P<0.05). And the CLP+ DPCPX+ Pae group’s values are obviously than the others(P<0.05). The values of IL-10, in contrast with them. Then the Sham groups are no sense of statistics between each other(P>0.05). 3. For cardiac tissue, all of the level of 6 cytokines in CLP groups are evidently higher than Sham groups(P<0.05). Except for IL-10, the CLP+ DMSO+ H2 O group’s values are remarkably higher than CLP+ DMSO + Pae group(P<0.05), but lower than CLP+ DPCPX+ H2 O group(P<0.05). And the C LP+ DPCPX+ Pae group’s values are obviously than the others(P<0.05). The values of IL-10, in contrast with them. Then the Sham groups are no sense of statistics between each other(P>0.05). 4. For, all of the dephosphorylation of P38, ERK and JNK in CLP groups are evidently higher than Sham groups(P<0.05). Except for Bcl-2, the CLP+DMSO+H2O group’s dephosphorylation are remarkably higher than C LP+DMSO + Pae group(P<0.05), but lower than C LP+DPCPX+H2O group(P<0.05). And the CLP+ DPCPX+ Pae group’s dephosphorylation are obviously than the others(P<0.05). The expression level of Bcl-2, in contrast with them. Then the Sham groups are no sense of statistics between each other(P>0.05).CONCLUSIONS: Paeoniflorin can inhabit their cardiac dysfunction and relieve myocardial injury in septic mice by regulating inflammatory cytokines in serum and cardiac tissue. The mechanisms of these role, may be intimately related to regulate the activation of MAPKs signaling pathway and the expression of Bcl-2, and simultaneously blocked by DPCPX. The results show that paeoniflorin has protective effects on cardiac dysfunction of septic mice through A1-adenosine receptor. |
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