三棱莪术配伍含药血清通过调控FABP4/PPARγ/NF-κB信号通路抑制子宫内膜间质细胞增殖、迁移和炎症的机制研究OA

Objective:Based on the fatty acid-binding protein 4(FABP4)/peroxisome proliferator-activated recep-tor γ(PPARγ)/nuclear factor κB(NF-κB)signaling pathway,this study investigated the effects of medicated serum from herbal pair SPARGANII RHIZOMA(SR)and CURCUMAE RHIZOMA(CR)on the proliferation,apoptosis,mi-gration,and inflammatory cytokine secretion of ectopic endometrial stromal cells(EESCs).Methods:Normal endometri-al tissues,as well as eutopic endometrium and ectopic lesions from patients with endometriosis(EMS),were collected clinically.The mRNA and protein expression levels of Fabp4 and Pparγ were detected using quantitative real-time PCR(qRT-PCR),immunofluorescence,and Western blot.Primary EESCs were cultured and incubated for 24 h with 10%blank serum,SR-medicated serum,CR-medicated serum,or equal-proportion SR-CR combination-medicated serum.Western blot was used to detect FABP4 protein expression.FABP4 knockdown and overexpression EESC models were established.Cells were divided into a blank control group(10%blank serum),FABP4 knockdown group,FABP4 over-expression group,and SR-CR combination-medicated serum group(10%).Cell viability was assessed using the CCK-8 assay.Apoptosis was determined by flow cytometry.The mRNA expression levels of Caspase3,Bcl2,and Bax were meas-ured by qRT-PCR.Cell migration was evaluated using a wound healing assay.Levels of interleukin(IL)-1β,IL-6,and tumor necrosis factor-α(TNF-α)in supernatants were measured by ELISA.Protein expression of PPARγ and phospho-rylation levels of NF-κB p65 were detected by Western blot.Results:Compared with normal eutopic endometrium,Fabp4 mRNA and protein expression levels were significantly upregulated in both eutopic endometrium and ectopic lesions of EMS,whereas Pparγ expression was significantly downregulated(P＜0.05).Compared with the blank control group,all treatment groups significantly downregulated FABP4 protein expression(P＜0.05).Compared with the blank control group,the FABP4 knockdown group showed significantly decreased cell viability,significantly increased apoptosis,signifi-cantly upregulated mRNA expression of Caspase3 and Bax,significantly downregulated Bcl2 expression,significantly re-duced migration rate,decreased IL-1β,IL-6,and TNF-α levels in supernatants,upregulated PPARγ protein expression,and reduced NF-κB p65 phosphorylation(P＜0.05).The FABP4 overexpression group showed opposite effects com-pared with the knockdown group(P＜0.05).Compared with the FABP4 overexpression group,the SR-CR combination-medicated serum group showed significantly decreased cell viability,increased apoptosis,upregulated Caspase3 and Bax mRNA expression,downregulated Bcl2 expression,reduced migration rate,decreased IL-1β,IL-6,and TNF-α levels,in-creased PPARγ protein expression,and decreased NF-κB p65 phosphorylation(P＜0.05).Conclusion:The equal-pro-portion combination of SR and CR can inhibit EESC proliferation,promote apoptosis,suppress migration,and reduce in-flammatory cytokine secretion by regulating the FABP4/PPARγ/NF-κB signaling pathway.