Abstract: Objective To explore the role of endoplasmic reticulum ryanodine receptor 1 (RyR1) expression and phosphorylation in sepsis-induced diaphragm dysfunction. Methods Thirty SPF male SD rats were randomized equally into 5 groups, including a sham-operated group, 3 sepsis model groups observed at 6, 12, or 24 h following cecal ligation and perforation (CLP; CLP-6h, CLP-12h, and CLP-24h groups, respectively), and a CLP-24h group with a single intraperitoneal injection of KN-93 immediately after the operation (CLP-24h+KN-93 group). At the indicated time points, diaphragm samples were collected for measurement of compound muscle action potential (CMAP), fatigue index of the isolated diaphragm and fitted frequency-contraction curves. The protein expression levels of CaMK II, RyR1 and P-RyR1 in the diaphragm were detected using Western blotting. Results In the rat models of sepsis, the amplitude of diaphragm CMAP decreased and its duration increased with time following CLP, and the changes were the most obvious at 24 h and significantly attenuated by KN-93 treatment (P<0.05). The diaphragm fatigue index increased progressively following CLP (P<0.05) irrespective of KN- 93 treatment (P>0.05). The frequency-contraction curve of the diaphragm muscle decreased progressively following CLP, and was significantly lower in CLP-24 h group than in CLP-24 h+KN-93 group (P<0.05). Compared with that in the sham-operated group, RyR1 expression level in the diaphragm was significantly lowered at 24 h (P<0.05) but not at 6 or 12 following CLP, irrespective of KN-93 treatment; The expression level of P-RyR1 increased gradually with time after CLP, and was significantly lowered by KN-93 treatment at 24 h following CLP (P<0.05). The expression level of CaMKII increased significantly at 24 h following CLP, and was obviously lowered by KN-93 treatment (P<0.05). Conclusion Sepsis causes diaphragmatic dysfunction by enhancing CaMK II expression and RyR1 receptor phosphorylation in the endoplasmic reticulum of the diaphragm.

Key words: sepsis; diaphragm dysfunction; RyR1; phosphorylation; muscle weakness