参莲提取物对M1巨噬细胞的影响

Effect on M1 macrophages of Shenlian extracts

该实验探讨参莲提取物对动脉粥样硬化发展中的M1巨噬细胞的影响。MTT法检测参莲提取物对RAW264.7细胞的毒性作用;γ干扰素(IFN-γ)及脂多糖(LPS)诱导RAW264.7细胞成M1型巨噬细胞,然后加入高、中、低剂量的参莲提取物(50,25,12.5 mg·L-1)作用,流式细胞术检测M1巨噬细胞膜分子CD86的表达,RT-PCR法检测M1巨噬细胞中iNOS和TNF-αmRNA的水平,ELISA法检测M1巨噬细胞上清中IL-6和TNF-α的含量。结果发现IFN-γ及LPS能诱导RAW264.7细胞极化为M1型巨噬细胞,M1巨噬细胞膜分子CD86的表达、细胞中iNOS和TNF-αmRNA的表达以及细胞分泌的IL-6和TNF-α的含量均有显著性地提高;参莲提取物各剂量均可抑制M1型巨噬细胞膜分子CD86、细胞中iNOS及TNF-αmRNA的表达,降低细胞分泌炎症细胞因子IL-6及TNF-α的含量。通过以上研究表明IFN-γ及LPS能诱导RAW264.7细胞极化为M1型巨噬细胞;参莲提取物对M1型巨噬细胞有一定的抑制作用。

This study discusses the effects of Shenlian extracts （SL） on M1 macrophages in atherosclerosis. The MTT assay was used to detect the growth inhibition rates of RAW264.7 cells. RAW264.7 cells were stimulated with murine interferon-γ （IFN-γ） plus lipopolysaccharide （LPS） to induce M1 macrophages. The different concentrations of SL extracts （high-dose 50 mg·L^-1 moderate-dose 25 mg·L^-1 low-dose 12.5 mg·L^-1 were added. The CD86 of M1 macrophages in cell membrane was measured by flow cytometry. The mRNA expression of iNOS and TNF-α gene was detected by reverse transcription PCR（RT-PCR）. And the supernatants were collected, the content of IL-6 and TNF-α were detected with ELISA kits. The results of this experiment show that the expression of the cell membrane molecule CD86, iNOS and TNF-α gene, the content of IL-6 and TNF-α was obviously increased in M1 macrophages by IFN-γ and LPS. The different doses of SL extract could reduce the expression of the above indicators. The above experimental results demonstrate that IFN-γ combined LPS can induce RAW264.7 cell to type into M1 macrophages, and SL extracts can inhibit M1 macrophages.