The Regulatory Effect Of SCFAs On Proliferation And Differentiation Of Bovine Preadipocytes

Fat deposition in animals is the product of proliferation and differentiation of preadipocytes.The increase and hypertrophy of adipocytes are closely related to fat deposition in animals.Short chain fatty acids(SCFAs)are involved in the regulation of lipid and glucose metabolism,and can regulate the balance between the synthesis and oxidation of fatty acids in the body.SCFAs also promoted adipocyte differentiation by reducing lipid accumulation and increasing the expression of peroxisome proliferator-activated receptor γ(PPARγ),CCAAT enhancer binding protein α(C/EBPα)and sterol regulatory element binding transcription factor 1(SREBF1).Therefore,we propose the scientific hypothesis that SCFAs may be involved in regulating the proliferation and differentiation of bovine preadipocytes,and we intend to explore the effect of SCFAs on the proliferation and differentiation of bovine preadipocytes in depth,and clarify the role of SCFAs in the proliferation and differentiation of bovine preadipocytes.This study provides a theoretical basis for the subsequent study of the effects of SCFAs on fat development and body fat deposition in cattle.In the present study,we isolated and cultured bovine preadipocytes in vitro to investigate the effect of SCFAs on the proliferation and differentiation of bovine preadipocytes.Different concentrations of SCFAs were added to bovine preadipocytes,and preadipocytes were divided into 4 groups.The cells were divided into Control group,Sodium acetate(SA)group,Sodium propionate(SP)group and Sodium butyrate(SB)group.At the end of the treatment,the cells were collected,and the proliferation of preadipocytes was detected by flow cytometry.The protein expression of cell proliferation-related protein Ki67 and the protein expression of cell cycle-related proteins,cyclin D1,D2,D3,E1(CCND1,CCND2,CCND3 and CCNE1)and cyclindependent kinases 2,4,6(CDK2,CDK4 and CDK6)were detected to identify the effect of SCFAs on proliferation of bovine preadipocytes.The same concentrations of SA,SP and SB were added during the process of differentiation into mature adipocytes.Oil red O staining was used to detect the formation of lipid droplets in adipocytes,and the protein expression levels of differentiation-related molecules including PPARγ,C/EBPα and SREBF1 in adipocytes were measured.Finally,the effect of SCFAs on the differentiation of bovine preadipocytes was determined.The results of in vitro experiments showed that 0.1 mmol/L SCFAs could promote the proliferation of bovine preadipocytes to varying degrees,increase the proliferation activity,accelerate the transition from G1 phase to S phase,and accelerate the cell cycle progression.At the same time,0.1 mmol/L SCFAs could promote the proliferation of preadipocytes by increasing the expression of proliferation-related protein Ki67 and cell cycle-related proteins CCND1,CCND2,CCND3,CCNE1,CDK2,CDK4 and CDK6.Under the stimulation of 0.1 mmol/L SCFAs,oil red O staining showed that the production of lipid droplets in bovine preadipocytes increased significantly and the lipid droplets became significantly larger.SCFAs can also promote the differentiation of bovine preadipocytes by up-regulating the protein expression of differentiationrelated proteins PPARγ,C/EBPα and SREBF1.In summary,in this study,bovine adipose precursor cells were isolated and cultured in vitro.All three different SCFAs,SA,SP and SB at 0.1mmol/L,promoted the proliferation and differentiation of bovine preadipocytes.This provides a theoretical basis for the subsequent study of the effects of SCFAs on fat development and body fat deposition in cattle,and has important theoretical and practical significance for the study of fat development and the regulation of body fat deposition in cattle.