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A **
16 5
** 4 *
mRNA level 10 mRNA level 3 2

p16 5 p21 1
0 0
6
Control 6 hh 12 h Control 6 hh 12 h
6
B
Leptin Leptin Leptin
Control
6 h 12 h **
* **
p16 16 kDa 1.5 * 1.5 ** 0.6 **
p16/β⁃actin 0.5 p21/β⁃actin 0.5 γ⁃H2AX/β⁃actin 0.2
p21 21 kDa 1.0 1.0 0.4
γ⁃H2AX 17 kDa 0 0 0
Control 6 h 12 h Control 6 h 12 h Control 6 h 12 h
β⁃actin 42 kDa
Leptin Leptin Leptin
C ** ** **
mRNA level 2.5 * mRNA level 4 3 2 mRNA level 5 4 3 2 * ** mRNA level 3 2 *
2.0
1.5
1.0
IL⁃1β 0.5 0 TNF⁃α 1 0 MCP⁃1 1 0 IL⁃6 1 0
Control 6 h 12 h Control 6 h 12 h Control 6 h 12 h Control 6 h 12 h
Leptin Leptin Leptin Leptin
D
Control Leptin **
β⁃GAL positive cell count 150

100
50

50 μm 50 μm 0 Control Leptin

A：The mRNA levels of p16 and p21 in leptin stimulated MCM. B：The protein expression of p16，p21，and γ⁃H2AX in leptin stimulated MCM.
**
*
C：The mRNA levels of IL⁃1β，TNF⁃α，MCP⁃1，and IL⁃6 in leptin stimulated MCM. D：Analysis of MCM senescence by β⁃Gal. P < 0.05，P < 0.01
（n=3）。
图1 瘦素促进小鼠心肌细胞衰老
Figure 1 Leptin promoted MCM senescence

Leptin treatment time（min）
* *
0 15 30 45 60 **
1.5 ** 1.0
p⁃PI3K 85 kDa * 0.8 **
p⁃PI3K/t⁃PI3K p⁃AKT/t⁃AKT 0.4
85 kDa 1.0
t⁃PI3K 0.6
p⁃AKT 60 kDa 0.5 0.2

60 kDa 0
t⁃AKT 0
0 15 30 45 60 0 15 30 45 60
β⁃actin 42 kDa Leptin treatment time（min） Leptin treatment time（min）

*
**
The phospholation levels of PI3K and AKT in leptin stimulated MCM. P < 0.05，P < 0.01（n=3）.
图2 瘦素激活小鼠心肌细胞PI3K/AKT信号通路
Figure 2 Leptin activated PI3K/AKT signaling pathway in MCM