马钱苷对AGEs致巨噬细胞极化的影响

目的 观察马钱苷对晚期糖基化终末产物（AGEs）致巨噬细胞极化的影响。方法 建立AGEs刺激巨噬细胞模型，设置空白对照组，模型组，氨基胍组，马钱苷低剂量组（1 μmol/L）和马钱苷高剂量组（10 μmol/L）。除空白对照组外，加入100 mg/L的AGEs刺激24 h后采用ELISA法检测细胞上清促炎性细胞因子IL-12、TNF-α和抑炎性细胞因子IL-10水平，采用流式细胞仪检测巨噬细胞M1型标记蛋白CD86和M2型标记蛋白CD206表达水平；免疫荧光法检测M1型标记蛋白iNOS和M2型标记蛋白CD206表达；Western blot法检测RAGE和巨噬细胞极化相关蛋白RBP-J、IRF8表达。结果 AGEs可上调IL-12、TNF-α水平（P<0.01），促进巨噬细胞iNOS、RAGE、RBP-J和IRF8蛋白表达（P<0.01）；而马钱苷预孵能够下调IL-12、TNF-α水平（P<0.01），上调IL-10水平，抑制RAGE、iNOS、RPB-J、IRF8蛋白表达（P<0.05~0.01）。结论 马钱苷可通过抑制RAGE/RBP-J/IRF8通路，抑制M1巨噬细胞极化，下调促炎因子IL-12、TNF-α水平，上调抑炎因子IL-10分泌，改善肾脏巨噬细胞炎症反应。 

Effects of Loganin on Macrophage Polarization Induced by AGEs

OBJECTIVE To investigate the effects of loganin on macrophage polarization induced by advanced glycation end products (AGEs). METHODS Macrophage injury model was established by AGEs stimulation， and control group， model group， aminoguanidine group， low-dose of loganin（1 μmol/L）and high-dose of loganin（10 μmol/L）groups were set. All groups were stimulated by 100 mg/L AGEs for 24 h after dosing， respectively， except control group. ELISA assay was used to detect the secretion of IL-10， IL-12 and TNF-α. Flow cytometry was employed to measure the expression of CD86 and CD206. The expression of iNOS and CD206 were detected by immunofluorescence assay. Western blot was used to detect the expression of RAGE， RBP-J and IRF8. RESULTS The secretion of IL-12， TNF-α and the expression of iNOS， RAGE， RBP-J and IRF8 were upregulated (P<0.01). After incubated with loganin， the secretion of IL-12 and TNF-α were downregulated while IL-10 was upregulated. The expression of CD86， iNOS， RAGE， RPB-J and IRF8 were decreased while the CD206 was increased after incubation with loganin (P<0.05， P<0.01). CONCLUSION RAGE/RBP-J/IRF8 pathway may be inhibited by loganin and the M1-type polarization may be prevented. The pro-inflammatory cytokines including IL-12 and TNF-α are down-regulated while anti-inflammatory cytokine IL-10 is up-regulated to alleviate the inflammatory injury in kidney.