Protective effect of CTRP6 on cerebral ischemia/reperfusion injury by attenuating inflammation, oxidative stress and apoptosis in PC12 cells

The newly identified C1q/tumor necrosis factor (TNF)-related protein-6 (CTRP6) is a highly conserved paralog of adiponectin with modulatory effects on metabolism and inflammation. However, the role of CTRP6 in cerebral ischemia/reperfusion (I/R) injury remains unknown. The aim of the present study was to explore the protective effects of CTRP6 against cerebral I/R injury and elucidate the possible underlying mechanisms. Oxygen-glucose deprivation and reperfusion (OGD/R) was used to induce an I/R injury model in vitro. Western blotting, reverse transcription-quantitative PCR, ELISA and flow cytometry analysis were used to measure the levels of CTRP6 along with those of inflammation-, oxidative stress- and apoptosis-related cytokines. The results indicated that CTRP6 expression was markedly downregulated following OGD/R. OGD/R also increased i) the activities of pro-inflammatory factors TNF-α, interleukin (IL)-1β, IL-6 and the levels of the oxidative products reactive oxygen species and malondialdehyde; ii) the ratio of apoptotic PC12 cells and iii) the expression of the pro-apoptotic proteins Bax, cleaved caspase-3 and cleaved caspase-9. In addition, the activities of the anti-inflammatory factors IL-10 and superoxide dismutase and the expression of the anti-apoptotic protein Bcl-2 were decreased. However, overexpression of CTRP6 rescued OGD/R-stimulated exacerbation of inflammation, oxidative stress and apoptosis. Mechanistically, OGD/R activated Ras homolog family member A (RhoA)/Rho-associated coiled-coil-containing protein kinase (Rock)/phosphatase and tensin homologue deleted on chromosome 10 (PTEN) signaling, whereas CTRP6 overexpression restored the expression of RhoA, Rock, PTEN, phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt). Furthermore, when CTRP6 and RhoA were overexpressed at the same time, RhoA abolished the protective effects of CTRP6 overexpression on OGD/R-induced inflammation, oxidative stress and apoptosis, while the presence of a PTEN inhibitor recovered the protective effects of CTRP6. Taken together, the findings of the present study indicate that CTRP6 attenuates cerebral ischemia/reperfusion-induced inflammation, oxidative stress and apoptosis via inhibiting the RhoA/Rock/PTEN pathway, thereby activating PI3K/Akt signaling.