梓醇对AGEs刺激巨噬细胞介导肾系膜细胞损伤的影响

Effect of catalpol on AGEs-stimulated RAW264.7 macrophage mediated mouse mesangial cell injury

目的探讨梓醇对晚期糖基化终末产物(AGEs)刺激巨噬细胞(RAW264.7)介导肾系膜细胞(MMCs)损伤的影响。方法将MMCs接种于Transwell小室,RAW264.7接种于下层孔板共培养,设置空白对照组、模型组、梓醇组(0.1、1.0、10.0μmol·L-1),设置氨基胍组(10.0μmol·L-1)作为阳性对照。各组加入药物孵育1 h后,用AGEs(100 mg·L-1)刺激RAW264.7,继续孵育23 h。采用MTT法检测MMCs增殖;免疫荧光法检测MMCs中COL-Ⅳ的表达;Western blot法检测MMCs中FN、COL-Ⅳ、TGF-β的表达;ELISA法检测RAW264.7上清液中IL-6、IL-12、TNF-α水平。结果梓醇可抑制AGEs刺激巨噬细胞介导系膜细胞的增殖(P<0.05,P<0.01),下调系膜细胞FN、COL-Ⅳ、TGF-β蛋白表达(P<0.05,P<0.01),降低巨噬细胞上清中IL-6、IL-12、TNF-α水平(P<0.05,P<0.01)。结论梓醇对AGEs刺激巨噬细胞介导系膜细胞损伤有明显的的保护作用,下调系膜细胞FN、COL-Ⅳ、TGF-β蛋白表达,抑制巨噬细胞炎症因子分泌,减轻炎症反应,从而缓解糖尿病肾病炎性损伤。

Aim To investigate the effect of catalpol on the stimulation of macrophage-mediated mouse mesangial cells(MMCs)injury by advanced glycation end products(AGEs).Methods MMCs and RAW264.7 macrophages were co-cultured in vitro and divided into control group,model group,catalpol group(0.1,1.0,10.0μmol·L-1),and aminoguanidine group(10.0μmol·L-1),which was setas the positive control.After drug administration for 1 h,RAW264.7 was stimulated by AGEs(100 mg·L-1)for 24 h.MTT assay was employed to detect the proliferation rate of MMCs.Immunofluorescence was used to investigate the expression levels of COL-IV in MMCs.Western blot was applied to assess the expression of FN,COL-IV and TGF-βin MMCs.ELISA was utilized to determine the levels of IL-6,IL-12 and TNF-αin supernatant liquid of RAW264.7 macrophages.Results Catalpol could inhibit RAW264.7 macrophage-mediated MMC-induced proliferation stimulated by AGEs(P<0.05,P<0.01),down-regulate the levels of FN,COL-Ⅳand TGF-βproteins in MMCs(P<0.05,P<0.01),and decrease IL-6,IL-12 and TNF-αlevel(P<0.05,P<0.01).Conclusions Catalpol has apparent protective effect on AGEs-stimulated macrophage-mediated mesangial cell injury.It down-regulates FN,COL-IV and TGF-βprotein expression in MMCs,down-regulates inflammatory factor levels and reduces inflammation,thereby alleviating diabetic kidney damage.