1.南京中医药大学养老服务与管理学院,江苏南京
2.南京市中医院风湿病科,江苏南京
WU Suling, E-mail:syf039@njucm.edu.cn
Received:06 August 2025,
Revised:2025-11-26,
Accepted:27 November 2025,
Published:15 December 2025
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华庆妮,陈月月,吴素玲.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[J].陆军军医大学学报,2025,47(23):2913-2921.DOI:10.16016/j.2097-0927.202508016.
HUA Q N,CHEN Y Y,WU S L.Ophiopogonis polysaccharides inhibit TNF-α/NF-κB/NLRP3 pathway to mitigate submandibular gland inflammation in a Sjögren’s syndrome murine model[J].J Army Med Univ,2025,47(23):2913-2921.DOI:10.16016/j.2097-0927.202508016.
华庆妮,陈月月,吴素玲.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[J].陆军军医大学学报,2025,47(23):2913-2921.DOI:10.16016/j.2097-0927.202508016. DOI:
HUA Q N,CHEN Y Y,WU S L.Ophiopogonis polysaccharides inhibit TNF-α/NF-κB/NLRP3 pathway to mitigate submandibular gland inflammation in a Sjögren’s syndrome murine model[J].J Army Med Univ,2025,47(23):2913-2921.DOI:10.16016/j.2097-0927.202508016. DOI:
目的
2
探究麦冬多糖(
Ophiopogon japonicus
polysaccharides, OJP)对干燥综合征(Sjögren’s syndrome,SS)小鼠颌下腺炎症的抗炎作用及其对肿瘤坏死因子-α(tumor necrosis factor,TNF-α)/核因子-κB(nuclear factor kappa-B,NF-κB)/NOD样受体热蛋白结构域 3(NOD like receptor family pyrin domain containing 3,NLRP3)通路的影响。
方法
2
①8只C57BL/6J小鼠设为空白组(Control组),32只C57BL/6J小鼠采用免疫诱导法建立SS模型,采用随机数字法分为(
n
=8):模型组(SS组)、低剂量麦冬多糖组[OJP(10 mg/kg)组]、高剂量麦冬多糖组[OJP(20 mg/kg)组]、羟氯喹组(Hyd组)。Control组、SS组灌服生理盐水,OJP(10 mg/kg)和OJP(20 mg/kg)组分别灌胃0.2 mL的10、20 mg/kg OJP溶液,Hyd组灌胃给予0.2 mL的100 mg/kg羟氯喹溶液,每日1次,共4周。测量小鼠饮水量及唾液流率,采用ELISA法检测小鼠血清中白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)、TNF-α炎症因子指标,HE染色观察各组小鼠颌下腺组织病理的改变,Western blot检测TNF-α/NF-κB/NLRP3通路相关蛋白的表达,免疫荧光法检测TNF-α、NLRP3蛋白表达。②将RAW264.7巨噬细胞分为空白对照组(Control组)、模型组(SS组)、麦冬多糖低剂量组(OJP1组)、麦冬多糖中剂量组(OJP2组)、麦冬多糖高剂量组(OJP4组),培养24 h后,去除培养基,Control组加入2 mL的DMEM完全培养基,SS组加入2 mL的含16 μg 脂多糖(lipopolysaccharides,LPS)的DMEM完全培养基,OJP不同剂量组分别加入2 mL的含16 μg LPS及浓度为1、2或4 mg/mL OJP的DMEM完全培养基。采用ELISA法检测炎症因子指标,Western blot检测TNF-α/NF-κB/NLRP3通路相关蛋白的表达,免疫荧光法检测TNF-α蛋白表达。
结果
2
①与空白组比较,SS组小鼠饮水量增加,唾液流率减少(
P
<
0.05),血清中炎症因子IL-1β、IL-6及TNF-α增多(
P
<
0.01),颌下腺组织HE染色显示病理损伤加重,NLRP3、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)、半胱天冬酶-1(cysteinyl aspartate specific proteinase,Caspase-1)、IL-1β、TNF-α、肿瘤坏死因子受体1(TNF receptor type 1,TNFR1)/甘油醛-3-磷酸脱氢酶(glyceraldehyde-3-phosphate dehydrogena
se ,GAPDH)及磷酸化NF-κB p65(p-NF-κB p65)/NF-κB p65升高(
P
<
0.01),免疫荧光结果显示TNF-α、NLRP3蛋白的表达升高。与SS组相比,Hyd组、OJP各剂量组小鼠饮水量减少,唾液流率增加(
P
<
0.01)、血清中炎症因子IL-1β、IL-6及TNF-α不同程度减少、颌下腺病理损伤均有减轻、Western blot结果显示NLRP3、ASC、Caspase-1、IL-1β、TNF-α、TNFR1/GAPDH及p-NF-κB p65/NF-κB p65蛋白表达降低(
P
<
0.01)、免疫荧光结果显示TNF-α、NLRP3蛋白的表达均有降低。②与单纯用LPS处理的巨噬细胞相比,经OJP干预后的细胞中的炎症因子、NLRP3、ASC、Caspase-1、IL-1β、TNF-α、TNFR1/GAPDH及p-NF-κB p65/NF-κB p65均有所降低(
P
<
0.01)。
结论
2
OJP可能通过抑制TNF-α/NF-κB/NLRP3信号通路激活达到抗炎作用,以此减轻SS小鼠炎症反应。
Objective
2
To investigate the anti-inflammatory effects of
Ophiopogon japonicus
polysaccharides (OJP) on submandibular gland inflammation in a mouse model of Sjögren’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
2
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
2
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
2
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.
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