Effect Of Berberine On Lipopolysaccharide-induced Inflammation Of Human Endometrial Stromal Cells

Objective: Endometritis is an endometrial inflammatory disease caused by pathogenic microbial infection.The main pathogens are common bacteria such as Escherichia coli,Streptococcus,followed by Mycoplasma.Berberine(BBR),also known as berberine,is an isoquinoline alkaloid that has long been considered a traditional medicine for clearing away heat and detoxifying and anti-intestinal bacterial infections.There are no reports on the study of berberine in the treatment of human endometritis.In view of its broad-spectrum antibacterial effect,this experiment detects the therapeutic effect of berberine by establishing an in vitro human endometrial stromal cell inflammation model.The anti-inflammatory mechanism of BBR is studied at the level.Methods: In this study,primary cultured cells were used to extract human endometrial stromal cells(hESCs)by secondary mesh screening.On the basis of this,a lipopolysaccharide(LPS)-induced hESCs-induced inflammation model was established.The CCK8 method was used to detect the proliferative effect of different concentrations of LPS and BBR on hESCs at different time points,and the LPS-induced inflammation model was identified by ELISA.In order to study the anti-inflammatory effect of BBR in vitro,the secretion of inflammatory factors IL-1βand TNF-α was detected by ELISA,and the mRNA expression levels of inflammatory factors IL-1β,TNF-α and IL-6 were detected by qRT-PCR.qRT-PCR was used to detect the mRNA expression of NF-κB classical inflammatory pathway-related proteins TLR4,IKKβ and NF-κB p65,and the protein expression of NF-κB p65 was detected by Western-Blot method.Results: 50μg/mL LPS measured by CCK8 method had the most proliferative effect on hESCs.There was no inhibition on hESCs in the range of ≤40μg/mL BBR.After induction of hESCs at 50μg/mL LPS for 24 h,the supernatant IL-1β wasdetected by ELISA.The level of TNF-α was significantly higher than that of the control group(P<0.001),which proved that the inflammatory model was established successfully.BBR inhibited the secretion of hESCs-induced inflammatory factors IL-1β and TNF-α induced by LPS,and down-regulated IL-1β,TNF-α and mRNA transcription level of IL-6(P<0.05);anti-inflammatory mechanism of BBR,BBR down-regulates LPS-induced mRNA expression of TLR4/IKKβ/NF-κB p65 inflammatory signaling pathway-related proteins TLR4,IKKβ,NF-κB p65(P<0.005),Western-Blot method showed that BBR inhibited the protein expression of NF-κB p65(P<0.005).Conclusion: BBR can down-regulate the gene expression levels of NF-κB inflammatory signaling pathway-related proteins TLR4,IKKβ,NF-κB p65 induced by LPS,as well as inhibit the expression of the key protein NF-κB p65 and reduce the expression of inflammatory factors(IL-1β,TNF-α,IL-6)and secretion(IL-1β,TNF-α),indicating that BBR can improve LPS-induced inflammation of human endometrial stromal cells,providing a new treatment for clinical treatment of endometritis.