金银花含药血清对脂多糖诱导的人牙周膜细胞活性及NLRP3/IL-1β通路的影响  

Effect of flos lonicerae medicated serum on the activity of LPS induced human periodontal membrane cells and the mechanism of NLRP3/IL-1β pathway

作　　者：陆平 岳黎[1] 魏丛丛[1] Ping LU;Li YUE;Cong-Cong WEI(Department of Stomatology,Liaocheng People's Hospital,Liaocheng 252000,Shandong Province,China)

机构地区：[1]聊城市人民医院口腔科,山东聊城252000

出　　处：《药物流行病学杂志》2023年第9期1017-1025,共9页Chinese Journal of Pharmacoepidemiology

摘　　要：目的探讨金银花含药血清对脂多糖(LPS)诱导的人牙周膜细胞(HPDLCs)活性及核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、白细胞介素1β(IL-1β)通路的影响。方法将20只大鼠按随机数字表法分为对照组(生理盐水)和金银花组(5.0 g·kg^(-1)),每组10只,灌胃给药,1次/d,持续14 d。将HPDLCs分为对照组(空白血清培养)、LPS组(10μg·mL^(-1)LPS+空白血清培养)、金银花低浓度(HPDLCs+10μg·mL^(-1)LPS+75μL空白血清培养+75μL 5%金银花含药血清)、中浓度(HPDLCs+10μg·mL^(-1)LPS+75μL空白血清培养+75μL 10%金银花含药血清)、高浓度(HPDLCs+10μg·mL^(-1)LPS+75μL空白血清培养+75μL 20%金银花含药血清)组。MTT检测HPDLCs细胞增殖率,Transwell小室检测HPDLCs细胞迁移,流式细胞仪检测HPDLCs细胞凋亡率,PCR检测HPDLCs中IL-1β、NLRP3 mRNA的相对表达;Western blotting法检测各组HPDLCs中IL-1β、NLRP3、含半胱氨酸的天冬氨酸蛋白水解酶1(Caspase-1)蛋白表达。结果与对照组比较,LPS组HPDLCs在24,48,72 h的细胞增殖率及细胞迁移数量显著降低(P<0.05),细胞凋亡率、IL-1β、NLRP3 mRNA和IL-1β、NLRP3、Caspase-1蛋白表达显著升高(P<0.05);与LPS组比较,金银花各剂量组HPDLCs在24,48,72 h的细胞增殖率和细胞迁移数量显著升高(P<0.05),细胞凋亡率、IL-1β、NLRP3 mRNA和IL-1β、NLRP3、Caspase-1蛋白表达显著降低(P<0.05)。结论金银花对改善LPS诱导的HPDLCs凋亡和促进HPDLCs的增殖和迁移具有一定作用,其机制可能与调控NLRP3、IL-1β及Caspase-1表达水平相关。Objective To investigate the effects of flos lonicerae containing serum on the activity of human periodontal membrane cells(HPDLCs)and NLRP3,IL-1β pathway induced by lipopolysaccharide(LPS).Methods Twenty rats were divided into controll group(normal saline)and flos lonicerae group(5.0 g·kg^(-1))based on a random number table method,with 10 rats in each group.HPDLCs cells were divided into control(CON)group(blank serum culture),LPS group(10μg·mL^(-1)LPS+blank serum culture),flos lonicerae low(HPDLCs+10μg·mL^(-1)LPS+75μL blank serum+75μL flos lonicerae containing serum),medium(HPDLCs+10μg·mL^(-1)LPS+75μL blank serum+75μL 10%flos lonicerae containing serum),high(HPDLCs+10μg·mL^(-1)LPS+75μL blank serum+75μL 20%flos lonicerae containing serum)concentration group was given.Proliferation rate of HPDLCs was detected by MTT,migration rate of HPDLCs was detected by Transwell chamber,apoptosis rate of HPDLCs was detected by flow cytometry,and relative expression of IL^(-1)βand NLRP3 in HPDLCs was detected by PCR.The expressions of IL-1β,NLRP3 and Caspase-1 protein containing cysteine in HPDLCs were detected by Western blotting.Results Compared with CON group,the proliferation rate at 24,48 and 72 h and migration number of HPDLCs in LPS group was significantly decreased(P<0.05),while the apoptosis rate,mRNA expression of IL-1βand NLRP3,and protein expression of IL-1β,NLRP3 and Caspase-1 were significantly increased(P<0.05).Compared with LPS group,the proliferation rate at 24,48 and 72 h and migration number of HPDLCs in flos lonicerae groups were significantly increased(P<0.05),while the apoptosis rate,mRNA expression of IL-1β,NLRP3,and protein expression of IL-1β,NLRP3 and Caspase-1 protein were significantly decreased(P<0.05).Conclusion Flos lonicerae can improve the apoptosis of HPDLCs and promote the proliferation and migration of HPDLCs induced by LPS,and the mechanism may be related to the regulation of NLRP3,IL-1βand Caspase-1 expression.

关 键 词：金银花 脂多糖 人牙周膜细胞 细胞活性 NOD样受体热蛋白结构域相关蛋白3/白细胞介素1β信号通路 

分 类 号：R285[医药卫生—中药学]