摘要
目的 研究茯苓酸(PA)对具核梭杆菌(Fn)感染的肠上皮细胞HIEC-6炎性反应的影响及其可能的作用机制。方法 将HIEC-6细胞分为对照组、模型组(1×10^(8) CFU·mL^(-1) Fn感染)、低剂量实验组(1×10^(8) CFU·mL^(-1) Fn+2μmol·L^(-1) PA)、中剂量实验组(1×10^(8) CFU·mL^(-1) Fn+4μmol·L^(-1)的PA)、高剂量实验组(1×10^(8) CFU·mL^(-1) Fn+8μmol·L^(-1)的PA)、pcDNA-TLR4组(1×10^(8) CFU·mL^(-1) Fn+8μmol·L^(-1)的PA+转染pcDNA-TLR4质粒)。处理12 h后分别用细胞计数试剂盒8(CCK-8)实验、Edu实验检测细胞增殖情况;分别用酶联免疫吸附测定(ELISA)实验和实时荧光定量聚合酶链反应(RT-qPCR)实验检测炎性细胞因子表达情况;用流式细胞术检测细胞凋亡情况;用蛋白质印迹法检测细胞相关蛋白表达水平。结果 对照组、模型组、低剂量实验组、中剂量实验组、高剂量实验组、pcDNA-TLR4组细胞凋亡率分别为(3.51±0.31)%、(23.38±1.85)%、(16.74±0.54)%、(12.10±1.44)%、(8.89±0.63)%和(14.87±0.88)%;白细胞介素-8(IL-8)含量分别为(23.25±1.41)、(79.69±4.37)、(68.94±2.63)、(51.46±2.56)、(34.18±2.63)和(71.09±5.08)pg·mL^(-1);白细胞介素-1β(IL-1β)含量分别为(8.81±0.49)、(51.65±2.13)、(35.98±3.47)、(24.83±2.47)、(14.79±1.25)和(43.89±1.59)pg·mL^(-1)。模型组与对照组比较,低、中、高剂量实验量组与模型组比较,pcDNA-TLR4组与高剂量实验组比较,在统计学上差异均有统计学意义(均P<0.05)。结论 茯苓酸可能通过抑制TLR4/NF-κB通路,抑制炎性反应改善Fn感染的肠上皮细胞凋亡。
Objective To investigate the effect of pachymic acid(PA)on HIEC-6 inflammatory response of intestinal epithelial cells infected by Fusobacterium nucleatum(Fn) and its possible mechanism.Methods HIEC-6 cells were divided into control group,model group(1×10^(8)CFU·mL^(-1) Fn infection),experimental-L group(1 × 10^(8) CFU·mL^(-1)Fn+2 μmol·L^(-1) PA),experimental-M group(1 × 10^(8)CFU·mL^(-1)Fn+4 μmol·L^(-1) PA),experimental-H group(1 × 10^(8) CFU·mL^(-1)Fn+8 μmol·L^(-1) PA),pcDNA-TLR4 group(1 ×10^(8) CFU·mL^(-1) Fn+8 μmol·L^(-1) PA+transfected pcDNA-TLR4).After 12 h of treatment,the cell proliferation was detected by cell counting kit-8(CCK-8) test and Edu test respectively.The expression of inflammatory cytokines was detected by enzyme-linked immunosorbent assay(ELISA) and real-time quantitative polymerase chain reaction(RT-qPCR) respectively.The apoptosis was detected by flow cytometry.Western blot assay was used to detect the expression level of cell related proteins.Results The apoptosis rates of control,model,experimental-L,experimental-M,experimental-H and pcDNA-TLR4 groups were(3.51±0.31)%,(23.38±1.85)%,(16.74±0.54)%,(12.10±1.44)%,(8.89±0.63)% and(14.87±0.88)%,respectively;IL-8 content were(23.25±1.41),(79.69±4.37),(68.94±2.63),(51.46±2.56),(34.18±2.63) and(71.09±5.08) pg · mL^(-1),respectively;IL-1β contents were(8.81±0.49),(51.65±2.13),(35.98±3.47),(24.83±2.47),(14.79±1.25) and(43.89±1.59) pg·mL^(-1).Compared between model group and control group,compared between experimental-L,experimental-M,experimental-H groups and model group,compared between pcDNA-TLR4 group and experimental-H group,the differences of the above indexes were all statistically significant(all P <0.05).Conclusion Pachymaric acid may improve the apoptosis of Fn infected intestinal epithelial cells by inhibiting TLR4/NF-κB pathway and inflammatory response.
作者
陈桂美
毛娟
CHEN Gui-mei;MAO Juan(School of Clinical Medicine,North Sichuan Medical University,Nanchong 637007,Sichuan Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2024年第4期544-548,共5页
The Chinese Journal of Clinical Pharmacology
关键词
茯苓酸
具核梭杆菌
肠上皮细胞
炎性反应
Toll-样受体4/核因子-κB通路
Pachymaric acid
Fusobacterium nucleatum
intestinal epithelial cells
inflammatory response
Toll-like receptor 4/nuclear factor-kB pathway
