文章摘要
化痰祛瘀法调控miRNA影响巨噬细胞抗动脉粥样硬化的分子机制
The molecular mechanism of the effect of phlegm and blood stasis regulating miRNA on macrophages against atherosclerosis
投稿时间:2020-06-12  录用日期:2020-07-15
DOI:
中文关键词: 动脉粥样硬化  化痰祛瘀法  miRNA-467b  巨噬细胞
英文关键词: Atherosclerosis  phlegm and blood stasis removing method  miRNA-467b  macrophages
基金项目:1.国家自然科学基金资助项目(No. 81704030),2.河南省中医药科学研究专项重点课题(No. 2019ZY1015),3.河南省中医药文化与管理研究项目(TCM2020016)。
作者单位邮编
张永晨* 驻马店市中医院 463000
秦合伟 河南省中医院 qinheweii@126.c
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中文摘要:
      目的 观察化痰祛瘀法是否通过miRNA-467b靶向LPL调控巨噬细胞,减少IL-6、IL-1β、TNF-α和MCP-1等炎性因子分泌和巨噬细胞脂质蓄积,研究化痰祛瘀法抗动脉粥样硬化的分子机制。方法 细胞实验,将RAW264.7巨噬细胞株随机分为5组,即对照组、模型组、miR-467b mimic组、中药高剂量含药血清组、中药低剂量含药血清组,不同组别的巨噬细胞经干预后,观察巨噬细胞增殖和凋亡情况;采用RT-PCR和Western-blot检测巨噬细胞中pri-miR-467b和LPL mRNA和蛋白表达;采用高效液相色谱法检测巨噬细胞内的胆固醇水平;采用ELISA法检测细胞培养基中的 IL-6、IL-1β、TNF-α和MCP-1等炎症因子水平。结果 含药血清干预后,中药高剂量组和低剂量组的巨噬细胞凋亡率和增殖率明显小于模型组(P<0.05);中药高剂量组和低剂量组巨噬细胞pri-miR-467b mRNA表达明显高于模型组(P<0.05);中药高剂量组和低剂量组巨噬细胞中LPL mRNA表达明显低于模型组(P<0.05);中药高剂量组和低剂量组巨噬细胞中LPL蛋白表达明显低于模型组(P<0.05);中药高剂量组和低剂量组巨噬细胞内的TC、CE水平明显低于模型组(P<0.05);中药高剂量组和低剂量组巨噬细胞内的FC水平明显高于模型组(P<0.05);中药高剂量组和低剂量组巨噬细胞培养基中IL-6、IL-1β、TNF-α和 MCP-1水平明显低于模型组(P<0.05);细胞实验各项检测指标在中药高剂量组和中药低剂量组间比较差异不明显(P>0.05)。结论 化痰祛瘀法抑制动脉粥样硬化斑块形成的作用机制可能与调控miRNA-467b靶向LPL调控巨噬细胞,减少IL-6、IL-1β、TNF-α和 MCP-1等炎性因子分泌和巨噬细胞脂质蓄积。
英文摘要:
      Objective To observe whether the phlegm and blood stasis treatment can target LPL to regulate macrophages through miRNA-467b, reduce the secretion of inflammatory factors such as IL-6, IL-1β, TNF-α and MCP-1 and the accumulation of lipid in macrophages. The molecular mechanism of anti-atherosclerosis with phlegm and blood stasis. Methods In cell experiments, the RAW264.7 macrophage cell line was randomly divided into 5 groups, namely control group, model group, miR-467b mimic group, high-dose Chinese medicine-containing serum group, low-dose Chinese medicine-containing serum group, and different groups. After intervention, macrophages were used to observe the proliferation and apoptosis of macrophages; RT-PCR and Western-blot were used to detect the expression of pri-miR-467b and LPL mRNA and protein in macrophages; high-performance liquid chromatography was used to detect macrophages. Cholesterol levels in phagocytes; ELISA method was used to detect the levels of IL-6, IL-1β, TNF-α, MCP-1 and other inflammatory factors in the cell culture medium. Results After drug-containing serum intervention, the apoptosis rate and proliferation rate of macrophages in the high-dose and low-dose groups of Chinese medicine were significantly less than the model group (P<0.05); macrophage pri-miR-467b in the high-dose and low-dose groups of Chinese medicine The mRNA expression was significantly higher than that in the model group (P<0.05); the expression of LPL mRNA in macrophages in the high-dose and low-dose groups of Chinese medicine was significantly lower than that in the model group (P<0.05); the macrophage in the high-dose and low-dose groups of traditional Chinese medicine Medium LPL protein expression was significantly lower than that in the model group (P<0.05); TC and CE levels in macrophages in the high-dose and low-dose groups were significantly lower than the model group (P<0.05); high-dose and low-dose groups in the traditional Chinese medicine The level of FC in macrophages of the group was significantly higher than that of the model group (P<0.05); the levels of IL-6, IL-1β, TNF-α and MCP-1 in the macrophage culture medium of the high-dose group and low-dose group of Chinese medicine were obvious It was lower than the model group (P<0.05); there were no significant differences in the cell test indicators between the high-dose Chinese medicine group and the low-dose Chinese medicine group (P>0.05). Conclusion The mechanism of phlegm and blood stasis inhibition of atherosclerotic plaque formation may be related to the regulation of miRNA-467b targeting LPL to regulate macrophages, reducing inflammatory factors such as IL-6, IL-1β, TNF-α and MCP-1 Secretion and lipid accumulation of macrophages.
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