Mechanism Of PPARγ Inhibiting TLR4 Expression In Mice Partial Hepatic Ischemia/reperfusion Injury

Test 1 Dynamic of rosiglitazone inhibition TLR4 expression in mouse partial live ischemia/reperfusionObjective To investigate the effect of rosiglitazone as PPARγagonist inhibiting the TLR4 expression in ischemic lobes in mouse partiallive ischemia/reperfusion injury (IRI)in BABL/C mice and explore the action of rosiglitazone inhibition the TLR4 receptor mediated inherent immune response.Mefhods The model of the mouse partial live IRI was established. All the animals were divided randomly into 3 groups: rosiglitazone+IRI group, vehicle (DMSO+IRI) group and sham group. The liver samples were collected when Mice were sacrificed after 0h, 2h, 4h and 6h to analyze the acute phase of liver IRI. The dynamic express of TlR4 mRNA was detected quantitatively by real-time quantitative RT-PCR. The level of TNF-αand ALT in portal vein were measured in all groups.Results After restoration of blood supply, the expression of TLR4 in ischemic lobes were strengthened in 0h, 2h, 4h, 6h of IRI in rosiglitazone+IRI group and vehicle group(P<0.05). The most intensive expression of TLR4 was present at 4 hour reperfusion in ischemic lobes in vehicle group. At the same time the expression of TLR4 mRNA in ischemic lobes in rosiglitazone group, as compared to vehicle group was significantly decrease (P<0.05). The expression of TLR4 of ischemic lobe at the 4h of blood supply restoration was the minimum among 0h, 2h, 4h, and 6h of blood supply restoration in rosiglitazone group (P < 0.05). The expression of TLR4 of ischemic lobe at the 4h of blood supply restoration was the maxlmum among 0h, 2h, 4h, and 6h of blood supply restoration in mice partial liver IRI in vehicle group (P< 0.05).The TNF-αand ALT of portal vein was down-regulated markedly in rosiglitazone group compared to vehicle group at each point of time in mice partial liver IRI (P<0.05).Conclusion Rosiglitazone inhibite the TLR4 receptor mediated inherent immune response and it may play roles in lessen the liver IRI. The best time of rosiglitazone inhibiting the TLR4 expression is four hours after reperfusion. Test 2 Mechanism of PPARγinhibiting TLR4 expression in mice partial hepatic ischemia/reperfusion injuryObjective:To investigate the mechanism and potential therapeutic targets for hepatic IRI, our study observed that PPAR-γregulate the TLR4 expression and portal vein serum levels of TNF-α, IL-10 and ALT in the mouse liver IRI. We research the interactive specific mechanism and biological effect of PPAR-γactivation by rosiglitazone, TLR4 and IL-10 in the process of hepatic partial IRI.Mefhods:The model of the mouse partial live IRI was established. All the animals were divided into 5 groups: rosiglitazone+IRI group, rosiglitazone+ BADGE+IRI group, BADGE +IRI group, DMSO+IRI group and sham group. The liver samples were collected when Mice were sacrificed after 4h to analyze the acute phase of liver IRI. PPARγand TLR4 mRNA was detected by real-time quantitative RT-PCR. The level of plasma TNF-α, IL-10 and ALT in portal vein were measured. Pathological sections of ischemic liver (HE staining) were used for histological analysis.Results:The expression of PPARγin rosiglitazone+IRI group was significantly heighten compared with rosiglitazone+ BADGE+IRI group ,BADGE+IRI group and DMSO+IRI group (P < 0.05). The expression of TLR4 mRNA in rosiglitazone+IRI group was significantly decrease compared with rosiglitazone+ BADGE+IRI group ,BADGE group+IRI and DMSO+IRI group (P<0.05). The level of IL-10 in portal vein was markedly up regulated in rosiglitazone+IRI group compared with rosiglitazone+ BADGE+IRI group ,BADGE+IRI group and DMSO+IRI group (P<0.05). The level of portal vein TNF-αand ALT was markedly down regulated in rosiglitazone+IRI group compared with rosiglitazone+ BADGE+IRI group ,BADGE group+IRI and DMSO+IRI group (P<0.05), above every group raise up the sham group(P<0.05). Pathology section revealed light to moderate liver cells edema in rosiglitazone+IRI group and liver cells severe edema, significant structural damage of lobular in rosiglitazone + BADGE +IRI group, BADGE+IRI group and DMSO+IRI group. Conclusion:These results demonstrated that rosiglitazone activating PPARγmarkedly depressed TLR4 expression and the level of portal vein TNF-α, along with increased the level of portal vein IL-10 through a PPARγ-dependent pathway. BADGE eliminate of the role rosiglitazone. PPARγmarkedly depressed TLR4 mediating inherent immune response to lead to lessen hepatocellular damage via IL-10 pathways.