妊娠期肝内胆汁瘀积症胎盘SIRT1异常表达及其意义研究

发布时间：2018-03-15 07:17

本文选题：白藜芦醇　切入点：SIRT1　出处：《重庆医科大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的检测炎症反应调节因子NF-κB及去乙酰化酶SIRT1在正常胎盘组织及ICP胎盘组织的蛋白表达情况;探讨牛磺胆酸(Taurocholic acid,TCA)诱导胎盘合体滋养细胞系HTR-8细胞的炎症反应,及白藜芦醇对HTR-8细胞炎症反应的抑制作用及机制。方法收集临床胎盘标本及肝功能指标(正常、ICP轻度和ICP重度),免疫组化及免疫印迹法检测SIRT1以及NF-κB的定位以及蛋白表达情况。不同浓度TCA处理HTR-8细胞24小时(组11:TCA=0μmol/L,组2:TCA=2μmol/L,组3:TCA=20μmol/L,组4:TCA=100μmol/L),免疫荧光及免疫印迹法检测SIRT1定位,SIRT1、NF-κB和TNF-α蛋白表达情况。SIRT1激动剂白藜芦醇(resveratrol,Res)处理各组细胞24小时,SIRT1抑制剂Ex-527与白藜芦醇同时处理各组细胞24小时,免疫印迹法检测处理后SIRT1、NF-κB及TNF-α蛋白表达的变化情况。结果SIRT1与NF-κB主要表达于胎盘合体滋养细胞的细胞质和细胞核。SIRT1蛋白在ICP胎盘组织的表达低于正常胎盘组织,NF-κB蛋白在ICP胎盘组织的表达高于正常胎盘组织;SIRT1蛋白在组3和组4中的表达低于组1和组2中的表达,NF-κB及TNF-α蛋白在组3和组4中的表达高于组1和组2中的表达。差异均具有统计学意义(P0.05)。使用白藜芦醇处理组3和组4后,SIRT1蛋白表达升高,NF-κB及TNF-α蛋白的表达降低(P0.05);Ex-527加白藜芦醇同时处理组3及组4,与单独使用白藜芦醇处理相比,SIRT1蛋白表达降低,NF-κB及TNF-α蛋白表达升高,差异均有统计学意义(P0.05)。结论ICP胎盘合体滋养细胞SIRT1表达下调,NF-κB及TNF-α表达上调,其可能的机制是ICP高浓度胆汁酸干扰胎盘合体滋养细胞SIRT1-NF-κB通路,诱导炎症反应,SIRT1激动剂白藜芦醇可逆转这一过程。
[Abstract]:Objective to detect the protein expression of NF- 魏 B and deacetylase SIRT1 in normal placenta and ICP placenta, and to investigate the inflammatory response induced by Taurocholic acidic acid (Taurocholic acidic acid) in human placental syncytiotrophoblast cell line HTR-8. Methods placental specimens and liver function indexes (mild and severe ICP) were collected, and the localization of SIRT1 and NF- 魏 B were detected by immunohistochemistry and Western blotting. Protein expression: HTR-8 cells were treated with different concentrations of TCA for 24 hours (group 11: TCA 0 渭 mol / L, group 2: TCA 2 渭 mol / L, group 3: TCA 20 渭 mol / L, group 4: TCA 100 渭 mol / L). Immunofluorescence and Western blotting were used to detect the expression of SIRT1 localization of SIRT1 NF- 魏 B and TNF- 伪 protein. Ex-527, an inhibitor of SIRT1, was treated with resveratrol for 24 hours. Results the expression of SIRT1 and NF- 魏 B in cytoplasm and nucleus of placenta syncytiotrophoblast was lower than that of normal placenta tissue in ICP placenta. The expression of TNF- 魏 B and TNF- 伪 protein in ICP placenta tissue was higher than that in normal placental tissue in group 3 and group 4. The expression of NF- 魏 B and TNF- 伪 protein in group 3 and group 2 was higher than that in group 1 and group 2. The expression of sir1 protein increased after resveratrol treatment group 3 and 4. The expression of NF- 魏 B and TNF- 伪 protein decreased in group 3 and group 4 after resveratrol treatment, and compared with resveratrol alone in group 3 and group 4. The expression of SIRT1 protein decreased and the expression of NF- 魏 B and TNF- 伪 protein increased. Conclusion the expression of SIRT1 in placental syncytiotrophoblast of ICP is down-regulated, and the expression of TNF- 魏 B and TNF- 伪 is up-regulated. The possible mechanism is that the high concentration of ICP bile acid interferes with the SIRT1-NF- 魏 B pathway of placental syncytiotrophoblast. Resveratrol, an agonist of SIRT1, can reverse this process.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R714.255

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