紫珠提取物通过抑制HMGB1水平改善小鼠宫颈上皮细胞炎症反应的机制  

Mechanism of Callicarpa nudiflora extract improving the inflammatory response of mice cervical epithelial cells by inhibiting the level of HMGB1

作　　者：段山红 关静 张丽花 DUAN Shan-hong;GUAN Jing;ZHANG Li-hua(Department of Gynecology,No.215 Hospital of Shaanxi Nuclear Industry,Xianyang Shaanxi 712000,China;Department of Obstetrics,Baoji High-tech Hospital,Baoji Shaanxi 721000,China)

机构地区：[1]陕西省核工业二一五医院妇科,陕西咸阳712000 [2]宝鸡高新医院产科,陕西宝鸡721000

出　　处：《临床和实验医学杂志》2022年第21期2245-2249,共5页Journal of Clinical and Experimental Medicine

基　　金：陕西省重点研发计划项目(编号:2019SF-209)。

摘　　要：目的探讨紫珠提取物通过抑制高迁移率族蛋白框1(HMGB1)水平改善小鼠宫颈上皮细胞炎症反应的机制。方法30只CD-1远交的孕鼠用于实验研究,通过脂多糖诱导早产小鼠模型。将成功怀孕的小鼠按照随机数字表法分成3组:对照组(正常孕鼠,等量0.9%氯化钠溶液灌胃,n=10),炎症模型组(早产模型,等量0.9%氯化钠溶液灌胃,n=10)和紫珠提取物组(早产模型,100 mg·kg^(-1)·d^(-1)紫珠提取物灌胃,n=10),共干预7 d。通过蛋白印迹分析小鼠宫颈组织HMGB1/TLR4/NF-κB的蛋白表达。通过酶联免疫吸附试验检测小鼠宫颈阴道粘液中细胞因子[肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-1β、IL-6、IL-8]的水平。通过RT-PCR分析小鼠宫颈组织焦亡相关蛋白iNOS、NLRP3和caspase-4的mRNA表达。通过水溶性荧光探针和可溶性E-钙粘蛋白(SECAD)水平评估上皮细胞通透性。结果炎症模型组HMGB1、TLR4和NF-κB的蛋白表达较对照组升高,紫珠提取物组HMGB1、TLR4和NF-κB的蛋白表达较炎症模型组降低,差异均有统计学意义(P<0.05)。炎症模型组TNF-α、IL-1β、IL-6和IL-8的水平较对照组升高,紫珠提取物组TNF-α、IL-1β、IL-6和IL-8的水平较炎症模型组降低,差异均有统计学意义(P<0.05)。炎症模型组iNOS、NLRP3和caspase-4的mRNA表达较对照组升高,紫珠提取物组iNOS、NLRP3和caspase-4的mRNA表达较炎症模型组降低,差异均有统计学意义(P<0.05)。炎症模型组SECAD水平和荧光强度较对照组升高,紫珠提取物组SECAD水平和荧光强度较炎症模型组降低,差异均有统计学意义(P<0.05)。结论脂多糖可以激活小鼠宫颈上皮细胞的炎症反应,这种炎症反应会导致宫颈上皮屏障受损,紫珠提取物通过抑制HMGB1表达降低小鼠宫颈上皮炎症反应,改善宫颈上皮屏障受损。Objective To explore the mechanism of Callicarpa nudiflora extract improving the inflammatory response of mice cervical epithelial cells by inhibiting the level of high mobility group box 1(HMGB1).Methods Thirty CD-1 outbred pregnant mice were used for experimental research.The preterm mice models were induced by lipopolysaccharide.The successfully pregnant mice were divided into three groups according to the random number table method:control group(normal pregnant mice,gavage with equal volume of normal saline,n=10),inflammation model group(preterm labor model,gavage with equal volume of normal saline,n=10)and Callicarpa nudiflora extract group(preterm labor model,100 mg·kg^(-1)·d^(-1)Callicarpa nudiflora extract gavage,n=10),a total of 7 days of intervention.The expressions of HMGB1/TLR4/NF-κB protein in mice cervical tissue were analyzed by Western blotting.Cytokine[tumor necrosis factor alpha(TNF-α),interleukin(IL)-1β,IL-6,IL-8]levels in mice cervicovaginal mucus were detected by enzyme-linked immunosorbent assay.The mRNA expressions of pyroptosis-related proteins iNOS,NLRP3 and caspase-4 in mice cervical tissue were analyzed by RT-PCR.Epithelial cell permeability was assessed by water-soluble fluorescent probes and soluble E-cadherin(SECAD)levels.Results The expressions of HMGB1,TLR4,and NF-κB protein in the inflammation model group were higher than those in the control group,and the expressions of HMGB1,TLR4,and NF-κB protein in the Callicarpa nudiflora extract group were lower than those in the inflammation model group,the differences were statistically significant(P<0.05).The levels of TNF-α,IL-1β,IL-6 and IL-8 in the inflammation model group were higher than those in the control group,the levels of TNF-α,IL-1β,IL-6 and IL-8 in the Callicarpa nudiflora extract group were lower than those in the inflammation model group,the differences were statistically significant(P<0.05).The expressions of iNOS,NLRP3 and caspase-4 mRNA in the inflammation model group were higher than those in the control grou

关 键 词：小鼠 紫珠提取物 高迁移率族蛋白框1 宫颈上皮细胞 炎症反应 

分 类 号：R285.5[医药卫生—中药学]