[1]刘　刚,崔朝初,卢　娜,等.复方心脉佳对巨噬细胞炎症相关因子caspase-1和白细胞介素-1β表达的影响及其机制[J].新乡医学院学报,2021,38(6):505-510.[doi:10.7683/xxyxyxb.2021.06.002]
　LIU Gang,CUI Chaochu,LU Na,et al.Effect of compound medicine Xinmaijia on the expression of inflammatory-related factors caspase-1 and interleukin-1β in macrophage and its mechanism[J].Journal of Xinxiang Medical University,2021,38(6):505-510.[doi:10.7683/xxyxyxb.2021.06.002]
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复方心脉佳对巨噬细胞炎症相关因子caspase-1和白细胞介素-1β表达的影响及其机制
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
38
期数:
2021年6
页码:
505-510
栏目:
基础研究
出版日期:
2021-06-05

文章信息/Info

Title:
Effect of compound medicine Xinmaijia on the expression of inflammatory-related factors caspase-1 and interleukin-1β in macrophage and its mechanism
作者:
刘　刚1; 崔朝初1; 卢　娜1; 冯超丽1; 刘　博1; 万光瑞2; 王现伟1
(1.新乡医学院基础医学院，河南省医用组织再生重点实验室，河南　新乡　453003；2.新乡医学院药学院，河南　新乡　453003)
Author(s):
LIU Gang1; CUI Chaochu1; LU Na1; FENG Chaoli1; LIU Bo1; WAN Guangrui2; WANG Xianwei1
(1.School of Basic Medicne,Xinxiang Medical University,Henan Key Laboratory of Medical Tissue Regeneration,Xinxiang 453003,Henan Province,China2.School of Pharmacy,Xinxiang Medical University,Xinxiang 453003,Henan Province,China)
关键词:
复方心脉佳; 动脉粥样硬化; 巨噬细胞; 白细胞介素-1β; caspase-1
Keywords:
compound medicine Xinmaijiaatherosclerosismacrophageinterleukin-1βcaspase-1
分类号:
R285
DOI:
10.7683/xxyxyxb.2021.06.002
文献标志码:
A
摘要:
目的　探讨复方心脉佳对巨噬细胞炎症相关因子caspase-1和白细胞介素-1β（IL-1β）表达的影响及其机制。方法　将对数生长期RAW264.7细胞随机分为对照组和体积分数0.5%、1.0%、2.5%、5.0%、10.0%、20.0%复方心脉佳组，应用细胞计数试剂盒检测复方心脉佳孵育24 h时各组细胞存活率。将对数生长期的RAW264.7细胞随机分为对照组、脂多糖（LPS）组和低、中、高剂量复方心脉佳组，对照组细胞正常培养不加任何刺激剂和药物，LPS组加入1 mg·L-1 LPS孵育4 h，低、中、高剂量复方心脉佳组分别加入体积分数1.0%、2.5%和5.0%的复方心脉佳预孵育12 h，随后加入1 mg·L-1 LPS处理4 h；应用Western blot检测5组细胞中 caspase-1前体、IL-1β前体蛋白相对表达量，实时荧光定量聚合酶链反应法检测对照组、LPS组、高剂量复方心脉佳组细胞中caspase-1前体、IL-1β前体及核转录因子κB (NF-κB）mRNA 相对表达量。将对数生长期THP-1细胞随机分为对照组、联合刺激组和低、中、高剂量复方心脉佳组，对照组细胞正常培养不加任何刺激剂和药物，联合刺激组细胞给予1 mg·L-1 LPS孵育4 h后再加入10 μmol·L-1尼日利亚菌素孵育1 h，低、中、高剂量复方心脉佳组分别加入体积分数1.0%、2.5%和5.0%的复方心脉佳预孵育12 h，然后加入1 mg·L-1 LPS孵育4 h后再加入10 μmol·L-1尼日利亚菌素孵育1 h；应用Western blot检测各组细胞培养液上清液中caspase-1和IL-1β相对表达量。结果　对照组与0.5%、1.0%、2.5%、5.0%、10.0%、20.0%复方心脉佳组RAW264.7细胞存活率分别为（100.00±2.43）%、（102.25±2.76）%、（102.05±2.76）%、（100.63±3.32）%、（94.17±2.05）%、（83.62±3.46）%、（79.44±9.60）%。低、中、高剂量复方心脉佳组RAW264.7细胞中caspase-1前体蛋白相对表达量显著低于对照组和LPS组（P<0.05），对照组与lps组raw264.7细胞中caspase-1前体蛋白相对表达量比较差异无统计学意义（>P>0.05）；LPS组和低、中、高剂量复方心脉佳组RAW264.7细胞中IL-1β前体蛋白相对表达量显著高于对照组（P<0.05），低、中、高剂量复方心脉佳组raw264.7细胞中il-1β前体蛋白相对表达量显著低于lps组（>P<0.05）；高剂量复方心脉佳组raw264.7细胞中caspase-1前体、il-1β前体蛋白相对表达量显著低于低、中剂量复方心脉佳组（>P<0.05），中剂量复方心脉佳组raw264.7细胞中caspase-1前体、il-1β前体蛋白相对表达量显著低于低剂量复方心脉佳组（>P<0.05）。lps组raw264.7细胞中caspase-1前体、il-1β前体和nf-κb mrna相对表达量显著高于对照组（P<0.05），高剂量复方心脉佳组raw264.7细胞中caspase-1前体、il-1β前体和nf-κb mrna相对表达量显著低于lps组和对照组（P<0.05）。联合刺激组和低、中、高剂量复方心脉佳组thp-1细胞上清液中caspase-1和il-1β蛋白相对表达量显著高于对照组（>P<0.05）；低、中、高剂量复方心脉佳组thp-1细胞上清液中caspase-1和il-1β蛋白相对表达量显著低于联合刺激组（>P<0.05）；高剂量复方心脉佳组thp-1细胞上清液中caspase-1和il-1β蛋白相对表达量显著低于低、中剂量复方心脉佳组（>P<0.05）；低剂量复方心脉佳组与中剂量复方心脉佳组thp-1细胞培养上清液中caspase-1和il-1β蛋白相对表达量比较差异无统计学意义（>P>0.05）。结论　复方心脉佳可能通过调控NF-κB信号通路而降低巨噬细胞中炎症相关因子caspase-1和IL-1β的表达和释放，其抗动脉粥样硬化功效可能与抗炎作用有关。undefinedundefinedundefinedundefinedundefinedundefinedundefinedundefinedundefinedundefined/i="">
Abstract:
Objective　To investigate the effect and mechanism of compound medicine Xinmaijia (XMJ) on the expression of inflammatory-related factors caspase-1 and interleukin-1β（IL-1β） in macrophage.Methods　　The RAW264.7 cells in logarithmic growth phase were randomly divided into control group and volume fraction of 0.5%,1.0%,2.5%,5.0%,10.0%,20.0% XMJ groups.The survival rate of RAW264.7 cells was detected by cell counting kit-8.The RAW264.7 cells in logarithmic growth period were randomly divided into the control group,lipopolysaccharide (LPS) group and low-,medium-,high-dose of XMJ groups.The cells in the control group were incubated with normal culture and not added with any stimulants and drugs,the cells in the LPS group were incubated with 1 mg·L-1 LPS for 4 hours,the cells in the low-,medium-,high-dose of XMJ groups were incubated with volume fraction of 1.0%,2.5% and 5.0% XMJ for 12 hours,respectively,and then they were treated with 1 mg·L-1 LPS for 4 hours;the relative expressions of pro-caspase-1 and pro-IL-1β protein in the five groups were detected by Western blot,the relative expressions of pro-caspase-1,Pro-IL-1β and nuclear transcription factors κB (NF- κB) mRNA in the control group,LPS group and high-dose of XMJ group were detected by real-time fluorescence quantitative polyme-rase chain reaction.The THP-1 cells in logarithmic growth phase were randomly divided into the control group,combined stimulation group and low-,medium-,high-dose of XMJ groups.The cells in the control group were incubated with normal culture and not added with any stimulants and drugs,the cells in the combined stimulation group were incubated with 10 μmol·L-1 nigericin for 1 hour after incubation with 1 mg·L-1 LPS for 4 hours,the cells in the low-,medium- and high-dose of XMJ groups were incubated with volume fraction of 1.0%,2.5% and 5.0% XMJ for 12 hours,and then incubated with 10 μmol·L-1 nigericin for 1 hour after incubation with 1 mg·L-1 LPS for 4 hours;Western blot was used to detect the relative expressions of caspase-1 and IL-1β in supernatant of THP-1 cells in each group.Results　The survival rate of RAW264.7 cells in control group and 0.5%,1.0%,2.5%,5.0%,10.0%,20.0% XMJ group were (100.00±2.43）%,（102.25±2.76）%,（102.05±2.76）%,（100.63±3.32）%,（94.17±2.05）%,（83.62±3.46）%,（79.44±9.60）%,respectively.The relative expression of pro-caspase-1 protein in RAW264.7 cells in the low-,medium- and high-dose of XMJ groups was significantly lower than that in the control group and LPS group (P<0.05),there was no significant difference in the relative expression of pro-caspase-1 protein in RAW264.7 cells between the control group and LPS group (P>0.05).The relative expression of pro-IL-1β protein in RAW264.7 cells in the LPS group and low-,medium-,high-dose of XMJ groups was significantly higher than that in the control group (P<0.05);the relative expression of pro-IL-1β protein in RAW264.7 cells in the low-,medium-,high-dose of XMJ groups was significantly lower than that in the LPS group (P<0.05).The relative expressions of pro-caspase-1 and pro-IL-1β protein in RAW264.7 cells in the high-dose of XMJ group were significantly lower than those in the low-,medium-dose of XMJ groups (P<0.05);the relative expressions of pro-caspase-1 and pro-IL-1β protein in the medium-dose of XMJ group were significantly lower than those in the low-dose of XMJ group (P<0.05).The relative expressions of pro-caspase-1,Pro-IL-1β and NF-κB mRNA in RAW264.7 cells in the LPS group were significantly higher than those in the control group (P<0.05);the relative expressions of pro-caspase-1,Pro-IL-1β and NF-κB mRNA in RAW264.7 cells in the high-dose of XMJ group were significantly lower than those in the control group and LPS group (P<0.05).The relative expressions of caspase-1 and IL-1β protein in supernatant of THP-1 cells in the combined stimulation group and low-,medium-,high-dose of XMJ groups were significantly higher than those in the control group (P<0.05);the relative expressions of caspase-1 and IL-1β protein in supernatant of THP-1 cells in the low-,medium-,high-dose of XMJ groups were significantly lower than those in the combined stimulation group (P<0.05);the relative expressions of caspase-1 and IL-1β protein in supernatant of THP-1 cells in the high-dose of XMJ group were significantly lower than those in the low-,medium-dose of XMJ groups (P<0.05);there was no significant difference in the relative expressions of caspase-1 and IL-1β protein in supernatant of THP-1 cells between the low-dose of XMJ group and medium-dose of XMJ group (P>0.05).Conclusion　XMJ may decrease the expression and secretion of inflammatory-related factors caspase-1 and IL-1β in macrophages through regulating NF-κB signaling pathway,the action of XMJ on atherosclerosis may be related to the anti-inflammatory effect.

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更新日期/Last Update: 2021-06-05