Visfatin Amplifies Cardiac Inflammation and Aggravates Cardiac Injury via the NF- κ B p65 Signaling Pathway in LPS-Treated Mice

Abstract

Background: Visfatin is an adipocytokine that has been demonstrated to be involved in cardiovascular diseases. This study aims at determining the role of visfatin in sepsis-induced cardiac injury and identify its possible mechanisms.

Methods: Dynamic changes in visfatin expression in mice with lipopolysaccharide- (LPS-) induced septicemia were measured. Additionally, mice were pretreated with visfatin and further administered LPS to observe the effects of visfatin on cardiac injury. Finally, septic mice were also pretreated with JSH-23 to investigate whether visfatin regulates cardiac injury via the NF-κB p65 pathway.

Results: Visfatin expression levels in both the heart and serum were increased in LPS-treated mice and peaked at 6 hours, and visfatin was derived from cardiac macrophages. In septic mice, pretreatment with visfatin reduced the survival rate, worsened cardiac dysfunction, and increased the expression of cardiac injury markers, including creatine kinase myocardial bound (CK-MB) and lactate dehydrogenase (LDH). Treatment with visfatin also increased the infiltration of CD3+ cells and F4/80+ cells, amplified the cardiac inflammatory response, and elevated myocardial cell apoptosis. Treatment with JSH-23 reversed the effects of visfatin in septic mice.

Conclusions: This study showed that visfatin amplifies the cardiac inflammatory response and aggravates cardiac injury through the p65 signaling pathway. Visfatin may be a clinical target for preventing cardiac injury in sepsis.

Figure 1

Expression of visfatin in septic mice. (a and b) Dynamic changes in visfatin levels in the serum and heart were detected in mice with LPS-induced septicemia. (c) Cardiac visfatin expression was detected in saline- and LPS-treated mice (200×). (d) Visfatin expression in cardiac macrophages was detected (200×). N = 5 in each group. ^∗p < 0.05 vs. the saline group. ^#p < 0.05 vs. the previous group.

Figure 2

Effects of visfatin on LPS-induced cardiac injury in mice. (a) The survival rates in the four groups were analyzed during follow up; N = 10 − 15 in each group. (b and c) The expression levels of CK-MB and LDH in both the serum and heart were examined. (d–f) LVEDD, LVESD, LVEF, FS, +dp/dt max, and −dp/dt max in each group were detected. N = 5 − 10 in each group. ^∗p < 0.05 vs. the saline + PBS group. ^#p < 0.05 vs. the LPS + PBS group. SL means saline; VFT means visfatin.

Figure 3

Effects of visfatin on cardiac inflammation in LPS-treated mice. (a) Cardiac CD3+ cells and CD68+ cells in the four groups were examined. (b) The expression of MCP-1, TNF-α, IFN-γ, IL-1β, IL-6, IL-12, IL-17, and IL-18 in the heart and serum was detected. (c) The activation of the NF-κB p65 pathway was measured. N = 5 − 6 in each group. ^∗p < 0.05 vs. the saline + PBS group. ^#p < 0.05 vs. the LPS + PBS group.

Figure 4

Effects of visfatin on LPS-induced myocardial cell apoptosis in mice. (a) The expression of cleaved caspase3 and Bcl2, apoptosis-related proteins, was detected. (b) TUNEL-positive cells were examined. N = 5−6 in each group. ^∗p < 0.05 vs. the saline + PBS group. ^#p < 0.05 vs. the LPS + PBS group. VFT means visfatin.

Figure 5

Effects of JSH-23 on LPS-induced cardiac injury and dysfunction in mice. (a) The effects of JSH-23 on the survival rates of visfatin-treated septic mice were measured, N = 10 − 15 in each group. (b and c) Cardiac injury markers in the serum and heart were measured. (d–f) Cardiac structure and function in each group were measured. N = 5 − 10 in each group. ^∗p < 0.05 vs. the LPS + DMSO + VFT group. VFT means visfatin.

Figure 6

Effects of JSH-23 on cardiac inflammation and myocardial cell apoptosis. (a) Phosphorylation of NF-κB p65 pathway components was detected. (b) Inflammatory mediators in the heart and serum were examined. (c). The numbers of TUNEL-positive myocardial cells were quantified. N = 5 − 10 in each group. ^∗p < 0.05 vs. the LPS + DMSO + VFT group.