RIPK3-Mediated Necroptosis in Diabetic Cardiomyopathy Requires CaMKII Activation

Figure 1

Cardiac dysfunction, CaMKIIδ activity, and necroptosis are augmented in DCM. Male C57BL/6 mice were injected with 60 mg/kg/d STZ for 5 consecutive days after a 12-hour overnight fast. Mice in the control group were injected with the same amount of citrate buffer. (a–c) Cardiac function was assessed by echocardiography, and EF, FS, and E/A were calculated. (d) Myocardium injury was measured by HE staining.  μm. (e) cTnI was detected. (f) The mRNA levels of CaMKIIδA, CaMKIIδB, and CaMKIIδC of the myocardium were detected by quantitative real-time PCR. 18S was serviced as a housekeeping mRNA. (g–h) Expression of CaMKII oxidation (ox-CaMKII), CaMKII phosphorylation (p-CaMKII), and total CaMKII was quantified by western blot. GAPDH was used as a loading control. (i) RIPK3 protein expression was quantified by western blot. GAPDH was used as a loading control. (j, k) Cell apoptosis of myocardium was detected with TUNEL staining and quantified with Image J analysis software.  μm. (l) Cleaved-caspase 3 and caspase 3 protein expression were quantified by western blot. GAPDH was used as a loading control. and , significantly from control, .

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