Inhibitory effect of complement 1q/tumor necrosis factor-related protein 4 on 3T3-L1 preadipocyte differentiation

Maireyanmu·Rozi; Wang Hongping; Zhang Cuiping; Xia Juan; Shen Tian; Lei Tao; Lu Jun; Gao Jie

doi:10.12307/2026.250

Abstract：BACKGROUND:Adipocyte differentiation is a complex biological process involving the transformation of preadipocytes into mature adipocytes.This process plays a key role in the development and progression of obesity and related metabolic diseases.In recent years,the complement C1q/tumor necrosis factor-related protein family,as a new member of the adipokine family,has become a research hotspot in the field of metabolic regulation. OBJECTIVE:To investigate the effect of complement 1q/tumor necrosis factor-related protein 4 on the differentiation of 3T3-L1 preadipocytes and its underlying mechanism. METHODS:The CCK-8 assay was used to determine the effects of different concentrations of complement 1q/tumor necrosis factor-related protein 4 on the viability of 3T3-L1 preadipocytes,and a safe concentration was selected for intervention.3T3-L1 preadipocytes were divided into a control group,an inducer group,and a complement 1q/tumor necrosis factor-related protein 4 group.3T3-L1 preadipocytes were first contact-inhibited for 2 days,followed by adipogenic inducer and complement 1q/tumor necrosis factor-related protein 4 treatment.On day 10 of differentiation,lipid droplet formation was observed using Oil Red O staining.The mRNA and protein expression levels of CCAAT/enhancer binding protein α,peroxisome proliferator-activated receptor γ,and fatty acid binding protein 4 were determined by RT-qPCR and western blot assay.Intracellular triglyceride and total cholesterol levels were measured using triglyceride and total cholesterol detection kits. RESULTS AND CONCLUSION:The safe maximum concentration of complement 1q/tumor necrosis factor-related protein 4 without toxicity to 3T3-L1 preadipocytes was 1 000 ng/mL.Compared with the control group,the inducer group showed increased intracellular lipid droplet formation,elevated triglyceride and total cholesterol levels(P<0.05),and significantly upregulated mRNA and protein expression levels of CCAAT/enhancer binding protein α,peroxisome proliferator-activated receptor γ,and fatty acid binding protein 4(P<0.05).Compared with the inducer group,the complement 1q/tumor necrosis factor-related protein 4 group showed reduced intracellular lipid droplet formation,lower triglyceride and total cholesterol levels(P<0.05).The mRNA and protein expression levels of CCAAT/enhancer binding protein α,peroxisome proliferator-activated receptor γ,and fatty acid binding protein 4 were significantly downregulated(P<0.05).These results suggest that complement 1q/tumor necrosis factor-related protein 4 inhibits the differentiation of preadipocytes into mature adipocytes,and its mechanism of action may be related to downregulating the expression of CCAAT/enhancer binding protein α,peroxisome proliferator-activated receptor γ,and fatty acid binding protein 4.