含3,4-二羟基的查尔酮类似物合成及对H2O2诱导PC12细胞损伤的保护作用

Primary investigation on synthesis of 3,4-dihydroxy chalcone analogues and its protective effect on H 2O 2-induced PC12 cell injury

目的合成含新型3,4-二羟基的查尔酮类似物并研究其对H 2O 2诱导PC12细胞损伤的保护作用。方法用不同的取代苯甲醛和3,4-二羟基苯乙酮为原料进行羟醛缩合反应,合成多种含有3,4-二羟基的查尔酮新类似物。采用噻唑蓝(MTT)法检测查尔酮类似物对PC12细胞的毒性作用,运用1,1-二苯基-2-三硝基苯肼(DPPH)清除实验检测查尔酮类似物直接清除自由基的作用;并用MTT法检测查尔酮类似物对H 2O 2诱导PC12细胞损伤的保护作用并筛选出活性较好的化合物5,运用免疫荧光实验和Western blot研究活性化合物5对核因子E2相关因子2(NRF2)抗氧化信号通路的作用。结果合成5个新型3,4-二羟基查尔酮类似物,并筛选出细胞毒性低且细胞保护作用较好的活性化合物5,该化合物对H 2O 2诱导PC12细胞的损伤具有良好的保护作用,呈现出浓度依赖性。活性化合物5可激活NRF2抗氧化信号通路,上调谷氨酸半胱氨酸合成酶催化亚基(GCLC)和血红素加氧酶-1(HO-1)蛋白的表达。结论新型活性化合物5对H 2O 2诱导PC12细胞氧化损伤表现出良好的保护作用且呈浓度依赖性,为得到具有较好活性的抗氧化查尔酮药物奠定了基础。

Objective To synthesize the new type 3,4-dihydroxy chalcone analogues and to study its protective effect on H 2O 2-induced PC12 cell injury.Methods Different substituted benzaldehydes and 3,4-dihydroxyacetophenone were used as the raw materials to conduct the aldol condensation reaction for synthesizing multiple new chalcone analogues containing 3,4-dihydroxy.The MTT assay was used to detect the cytotoxicity of chalcone analogues on PC12 cells.The DPPH assay was used to check the effect of chalcone analogues for directly scavenging the free radicals.Moreover the MTT assay was to detect the protective effect of chalcone analogues on H 2O 2-induced PC12 cell injury and screen the compound 5 with better activity.The effect of active compound 5 on the antioxidant signaling pathway of NRF2 was investigated by using Western-blot and immunefluorescence assay,respectively.Results The five new type 3,4-dihydroxychalcone analogues were synthesized,and the active compound 5 possessing low cytotoxicity and good cytoprotective effect was screened out,which had good protective effect on H 2O 2-induced damage of PC12 cells,showing a concentration-dependent manner.The active compound 5 could activate the NRF2 antioxidant signaling pathway and up-regulated the expression of GCLC and HO-1 proteins.Conclusion The new type active compound 5 shows a good protective effect on H 2O 2-induced oxidative damage of PC12 cells in a concentration-dependent manner,which laid a prophase foundation for obtaining anti-oxidant chalcone drugs with better activity.