Figure 7.

TNFα inhibition inhibits CKD‐dependent vascular calcification by inhibiting aortic PERK‐eIF2α‐ATF4‐CHOP signaling. A, Survival rate of 5/6 nx ApoE^−/− mice treated with either vehicle (saline) or 10 mg/kg infliximab. 17 animals per group were analyzed for survival analysis. B, Calcified area stained by von Kossa in aortic sinus of sham‐operated ApoE^−/− mice (left), 5/6 nephrectomized ApoE^−/− mice treated with vehicle (middle) or infliximab (right). Arrow indicates calcified lesion. C, Calcified lesions in 5/6 nx or sham‐operated ApoE^−/− mice treated with either infliximab or vehicle for 10 weeks. ***P<0.001 vs sham‐operated ApoE^−/− mice treated with vehicle. ^###P<0.001 vs 5/6 nx ApoE^−/− mice treated with vehicle. Six samples per group were analyzed. D, Aortic mRNA levels in 5/6 nx or sham‐operated ApoE^−/− mice treated with either infliximab or vehicle for 10 weeks. *P<0.05 vs sham‐operated ApoE^−/− mice treated with vehicle. ^#P<0.05 vs 5/6 nx ApoE^−/− mice treated with vehicle. Six samples per group were analyzed. E, Aortic, p‐PERK, (p)‐ and total ATF4, and CHOP protein expression in 5/6 nx or sham‐operated ApoE^−/− mice treated with either infliximab or vehicle. F, Immunohistochemical detection of CHOP in the aortic sinus of 5/6 nx ApoE^−/− mice treated with either infliximab or vehicle. G, CHOP‐positive area in 5/6 nx or sham‐operated ApoE^−/− mice treated with either infliximab or vehicle for 10 weeks. **P<0.01 vs sham‐operated ApoE^−/− mice treated with vehicle. ^##P<0.01 vs 5/6 nx ApoE^−/− mice treated with vehicle. Six samples per group were analyzed. ALP indicates alkaline phosphatase; ApoE, apoprotein E; ATF4, activating transcription factor 4; CHOP, C/EBP homologous protein; nx, nephrectomized; OCN, osteocalcin; TNFα, tumor necrosis factor‐α.