参红通络方调控巨噬细胞极化防治动脉粥样硬化机制

Mechanism of Shen-Hong-Tong-Luo formula in regulating macrophage polarization and preventing atherosclerosis

目的探讨参红通络方对ApoE-/-小鼠巨噬细胞极化的影响。方法将40只ApoE-/-小鼠随机分成模型组、参红通络低、中、高剂量组,每组各10只,C57BL/6小鼠10只作为空白对照组,适应性喂养1 w,ApoE-/-小鼠给予高脂饲料喂养7 w后,给予相应药物灌胃,C57BL/6小鼠普通饲料喂养7 w后,给予生理盐水灌胃。给药结束后,戊巴比妥麻醉取材,采用大体油红O染色及苏木素-伊红(HE)染色对小鼠主动脉整体和主动脉窦进行染色;免疫组化法检测M1型标志物CD86、M2型标志物CD206在小鼠主动脉根部病理组织切片中的表达情况;Western印迹法检测巨噬细胞极化作用相关蛋白诱导型一氧化氮合酶(iNOS)、精氨酸酶(Arg)-1的表达水平。结果与空白对照组比较,模型组主动脉整体、主动脉窦斑块占比均显著增加(P<0.05),主动脉组织中CD86、iNOS蛋白表达都明显增加(P<0.05),CD206、Arg-1蛋白表达均显著下降(P<0.05)。与模型组对比,参红通络各剂量组主动脉整体、主动脉窦斑块占比明显下降(P<0.05),主动脉中CD86、iNOS蛋白表达水平显著降低(P<0.05),而CD206、Arg-1蛋白表达水平显著升高(P<0.05)。结论参红通络方可通过抑制M1型巨噬细胞极化、促进M2型巨噬细胞极化,减少炎症反应,维持动脉内粥样斑块的稳定性,从而发挥抗动脉粥样硬化的作用。

Objective To investigate the effect of Shen-Hong-Tong-Luo formula on macrophage polarization in ApoE-/-mice.Methods Forty ApoE-/-mice were randomly divided into model group,Shen-Hong-Tong-Luo low,middle and high dose group,with 10 mice in each group.Ten C57BL/6 mice were used as blank control group,adaptive feeding for 1 week,ApoE-/-mice were fed with high-fat diet for seven weeks,and then were given corresponding drugs by gavage.C57BL/6 mice were fed with normal diet for seven weeks,and then were given physiological saline by gavage.After the administration,the whole aorta and aortic sinus were stained with oil red O and HE;the expressions of M1 marker CD86 and M2 marker cd206 were detected by immunohistochemistry;the expression levels of inducible nitric oxide synthase(iNOS)and arginase(Arg)-1 were detected by Western blot.Results Compared with control group,the proportion of aortic plaque and aortic sinus plaque in model group increased significantly(P<0.05),and the expression of CD86 and iNOS protein in aortic tissue was significantly increased(P<0.05),and the expression of CD206 and Arg-1 were significantly decreased(P<0.05).Compared with model group,the proportion of plaque of aorta and sinus of mice in all dose groups were decreased significantly(P<0.05),and the expression levels of CD86 and iNOS protein in the aorta of mice in each dose group were decreased(P<0.05),while the expression levels of CD206 and Arg-1 were increased(P<0.05).Conclusions Shen-Hong-Tong-Luo formula could inhibit the polarization of M1 macrophages,promote the polarization of M2 macrophages,reduce the inflammatory reaction,maintain the stability of atherosclerotic plaque,and thus play the role of against atherosclerosis.