论文部分内容阅读
目的探讨分形趋化因子(FKN)对H9C2心肌细胞炎症因子、细胞凋亡的影响。方法采用(100、200、300、400、500)ng/m L的可溶性FKN(s FKN)处理H9C2细胞0、12、24、36、48 h,利用MTT法检测s FKN对H9C2细胞增殖的作用。筛选出200 ng/m L s FKN作用H9C2细胞24 h后,实时定量PCR和Western blot法分别检测细胞肿瘤坏死因子α(TNF-α)mRNA和蛋白的表达,筛选出s FKN最佳处理剂量和处理时间。采用200 ng/m L s FKN,200 ng/m L s FKN与5 ng/m L AKT抑制剂LY294002分别处理H9C2细胞24 h,通过Hochest33258染色检测细胞核染色质聚集情况。采用实时定量PCR和Western blot法分别检测H9C2细胞中Bax和Bcl-2的表达。结果 s FKN可以抑制H9C2细胞的增殖情况。FKN促进心肌细胞TNF-α的表达,具有剂量依赖性。FKN可通过上调Bax蛋白的表达与下调Bcl-2蛋白的表达促进心肌细胞凋亡。FKN可能通过激活PI3K/AKT通路促进心肌细胞的炎症发生与凋亡。结论 FKN通过激活PI3K/AKT通路促进心肌细胞的炎症反应和凋亡作用。
Objective To investigate the effects of fractal chemokine (FKN) on the inflammatory cytokines and apoptosis in H9C2 cardiomyocytes. Methods H9C2 cells were treated with soluble FKN (sFKN) at (100, 200, 300, 400, 500 ng / mL) for 0,12,24,36,48 h. MTT assay was used to detect the effect of sFKN on the proliferation of H9C2 cells . The expression of TNF-α mRNA and protein were detected by real-time quantitative PCR and Western blot respectively after treated with 200 ng / m L s FKN for 24 h. The optimal dose of sFKN and Processing time. H9C2 cells were treated with 200 ng / m L s FKN, 200 ng / m L s FKN and 5 ng / m L AKT inhibitor LY294002 for 24 h, respectively. Hochest33258 staining was used to detect the nuclear chromatin accumulation. Real-time quantitative PCR and Western blot were used to detect the expression of Bax and Bcl-2 in H9C2 cells. Results s FKN can inhibit the proliferation of H9C2 cells. FKN promoted the expression of TNF-α in cardiomyocytes in a dose-dependent manner. FKN can promote cardiomyocyte apoptosis by up-regulating Bax protein expression and down-regulating Bcl-2 protein expression. FKN may promote cardiomyocyte inflammation and apoptosis by activating PI3K / AKT pathway. Conclusion FKN can promote the inflammatory response and apoptosis of cardiomyocytes by activating PI3K / AKT pathway.