Ophiopogonis polysaccharides inhibit TNF-α/NF-κB/NLRP3 pathway to mitigate submandibular gland inflammation in a Sj?gren's syndrome murine model
10.16016/j.2097-0927.202508016
- VernacularTitle:麦冬多糖通过抑制TNF-α/NF-κB/NLRP3信号通路减轻干燥综合征小鼠颌下腺炎症
- Author:
Qingni HUA
1
;
Yueyue CHEN
;
Suling WU
Author Information
1. 南京中医药大学养老服务与管理学院
- Keywords:
Ophiopogon japonicus polysaccharide;
Sj?gren's syndrome;
TNF-α/NF-κB pathway;
NOD like receptor family pyrin domain containing 3
- From:
Journal of Army Medical University
2025;47(23):2913-2921
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the anti-inflammatory effects of Ophiopogon japonicus polysaccharides(OJP)on submandibular gland inflammation in a mouse model of Sj?gren's syndrome(SS)and its impact on the tumor necrosis factor-α(TNF-α)/nuclear factor kappa-B(NF-κB)/NOD-like receptor family pyrin domain containing 3(NLRP3)signaling pathway.Methods ① Eight C57BL/6J mice served as the control group(Control group),while 32 C57BL/6J mice were subjected to immune induction to establish the SS model and then randomly divided into 4 groups(n=8 each):model group(SS group),low-dose OJP group[OJP(10 mg/kg)group],high-dose OJP group[OJP(20 mg/kg)group],and hydroxychloroquine group(Hyd group).The Control and SS groups received intragastric administration of saline,the OJP(10 mg/kg)and OJP(20 mg/kg)groups received 0.2 mL of OJP solution at 10 mg/kg or 20 mg/kg,respectively,and the Hyd group received 0.2 mL of hydroxychloroquine solution at 100 mg/kg,once daily for 4 weeks.Water consumption and salivary flow rate were measured.Serum levels of inflammatory cytokines interleukin-6(IL-6),interleukin-1β(IL-1β),and TNF-α were detected by ELISA.Pathological changes in submandibular gland tissue were observed by HE staining.Protein expression related to the TNF-α/NF-κB/NLRP3 pathway was assessed by Western blot,and TNF-α and NLRP3 protein expression was detected by immunofluorescence.② RAW264.7 macrophages were divided into blank control group(Control group),model group(SS group),low-dose OJP group(OJP1 group),medium-dose OJP group(OJP2 group),and high-dose OJP group(OJP4 group).After 24 hours of culture,the medium was removed;the Control group received 2 mL of complete DMEM medium,the SS group received 2 mL of complete DMEM medium containing 16 μg LPS,and the different OJP dose groups received 2 mL of complete DMEM medium containing 16 μg LPS plus OJP at concentrations of 1,2,or 4 mg/mL,respectively.Inflammatory cytokine levels were measured by ELISA,TNF-α/NF-κB/NLRP3 pathway-related protein expression was detected by Western blot,and TNF-α protein expression was detected by immunofluorescence.Results ① Compared with the Control group,the SS group showed increased water consumption and decreased salivary flow rate(P<0.05),elevated serum levels of IL-1β,IL-6,and TNF-α(P<0.01),aggravated pathological damage observed by HE staining in submandibular glands,and increased expression of NLRP3,apoptosis-associated speck-like protein containing a CARD(ASC),cysteinyl aspartate specific proteinase(Caspase-1),IL-1β,TNF-α,TNF receptor type 1(TNFR1)/glyceraldehyde-3-phosphate dehydrogenase(GAPDH),and phosphorylated NF-κB p65(p-NF-κB p65)/NF-κB p65(P<0.01),with immunofluorescence also showing elevated TNF-α and NLRP3 protein expression.Compared with the SS group,the Hyd group and both OJP dose groups exhibited decreased water consumption,increased salivary flow rate(P<0.01),reduced serum levels of IL-1β,IL-6,and TNF-α,alleviated submandibular gland pathological damage,decreased expression of NLRP3,ASC,Caspase-1,IL-1β,TNF-α,TNFR1/GAPDH,and p-NF-κB p65/NF-κB p65 proteins by Western blotting(P<0.01),and reduced TNF-α and NLRP3 protein expression by immunofluorescence.② Compared with macrophages treated solely with LPS,those treated with OJP intervention showed reduced levels of inflammatory cytokines and decreased expression of NLRP3,ASC,Caspase-1,IL-1β,TNF-α,TNFR1/GAPDH,and p-NF-κB p65/NF-κB p65(P<0.01).Conclusion OJP may exert anti-inflammatory effects by inhibiting the activation of the TNF-α/NF-κB/NLRP3 signaling pathway,thereby alleviating the inflammatory response in SS mice.
