利拉鲁肽抗CXC趋化因子配体16诱导的人足细胞脂质沉积作用及机制

Objective To explore the effect and mechanism of liraglutide on CXC chemokine ligand 16 (CXCL16)-induced lipid deposition in human podocytes. Methods ① Glucose 40 mmol·L-1 stimulated human podocytes for 24 h, and palmitic acid 250 μmol·L-1 stimulated human podocytes for 6 h. The expression level of CXCL16 protein in podocytes was detected by Western blotting. ② Recombinant CXCL16 100 μg·L-1 stimulated podocytes for 0, 6, 12 and 24 h, and Western blotting was used to detect the expression level of podocyte schizin protein. ③ The podocytes were divided into cell control group, CXCL16 100 μg·L-1 group, CXCL16+liraglutide 10, 50 and 100 nmol·L-1 group, and CXCL16+simvastatin 100 nmol·L-1 group, adding 2 drugs After 1 h, recombinant CXCL16 100 μg·L-1 was added and cultured for 24 h, and the lipid droplet area of ​​podocytes was detected by Oil Red O staining. ④ The podocytes were divided into cell control group, CXCL16 100 μg·L-1 group, CXCL16+liraglutide 100 nmol·L-1 group, and CXCL16+simvastatin 100 nmol·L-1 group. CXCL16 was added 2 hours after the drug was added. 100 μg·L-1 was continued to be cultured for 24 h. Phalloidin staining was used to detect the percentage of actin stress fibers in podocytes. Tumor necrosis factor α (TNF-α) and transforming growth factor β (TGF-β) in the cell culture medium were detected by ELISA. ) and interleukin 1β (IL-1β) protein concentration, real-time fluorescence quantitative PCR was used to detect TNF-α, TGF-β and IL-1β mRNA expression levels; Western blotting was used to detect podocyte cleavage factor and cholesterol regulatory element binding protein 1 ( SREBP1), SREBP2 and adipogenic differentiation-related protein (ADRP) expression levels. Results ① Compared with the cell control group, both high sugar and palmitic acid stimulation caused a significant increase in the expression level of CXCL16 in human podocytes (P<0.01). ② Compared with the cell control group, CXCL16 stimulation can cause a significant down-regulation of podocyte schizophrenia expression (P<0.01). ③ Compared with the cell control group, CXCL16 stimulation can cause a significant increase in the lipid droplet area of ​​podocytes (P<0.01), and both simvastatin and different concentrations of liraglutide can significantly alleviate this change (P<0.01). ④ Compared with the cell control group, recombinant CXCL16 can cause a significant decrease in the percentage of actin stress fibers in podocytes (P<0.01), and a significant increase in the protein concentrations of TNF-α, TGF-β and L-1β in the culture medium (P< 0.01), podocyte TNF-α, TGF-β and IL-1β mRNA levels were significantly increased (P<0.05, P<0.01), schizolin expression was significantly down-regulated (P<0.01), SREBP1, SREBP2 and ADRP protein expression levels Significantly increased (P<0.01); compared with the CXCL16 group, liraglutide and simvastatin could significantly restore the percentage of actin stress fibers in podocytes (P<0.01) and downregulate TNF-α, TGF-β and TNF-β in the culture medium. IL-1β protein concentration (P<0.01), inhibited podocyte TNF-α, TGF-β and IL-1β mRNA levels (P<0.05, P<0.01), up-regulated fissin protein expression level (P<0.01), down-regulated SREBP1, SREBP2 and ADRP protein expression levels (P<0.01). Conclusion Liraglutide can reduce podocyte damage by inhibiting lipid deposition induced by CXCL16 and alleviate inflammatory activation caused by lipid deposition, which may be related to liraglutide's ability to inhibit the expression of SREBP and ADRP.