晚期糖基化终产物受体特异性小干扰RNA对致纤维化细胞因子生成的抑制作用

发布时间：2018-03-22 16:20

本文选题：小干扰RNA　切入点：糖基化终产物受体　出处：《医学研究生学报》2013年05期 　论文类型：期刊论文

【摘要】：目的晚期糖基化终产物受体(the receptor of advanced glycation end products,RAGE)特异性小干扰RNA(small in-terfering RNA,siRNA)能在肝星状细胞(hepatic stellate cell,HSCs)T6内抑制RAGE、α-平滑肌肌动蛋白(alpha-smooth muscle ac-tin,α-SMA)和Ⅰ型胶原mRNA和蛋白的表达,其在HSCs的激活和胶原的形成中发挥着重要的作用。探讨RAGE特异性siRNA在原代大鼠HSCs中对致纤维化细胞因子生成的影响。方法构建RAGE特异性siRNA表达载体,分离培养原代大鼠HSCs,将重组载体转染入原代大鼠HSCs,以空白组和转染非特异性siRNA表达载体pAKD-NC组为对照,分别用PCR法和Western blot检测各组原代HSCs RAGE、β1转化生长因子(transforming growth factor-β1,TGF-β1)和结缔组织生长因子(connective tissuegrowth factor,CTGF)mRNA及蛋白的表达。结果转染特异性siRNA表达载体pAKD-GR126的原代HSCs RAGE mRNA和相对分子质量为42000的RAGE蛋白的表达分别为空白组和pAKD-NC组的(42.32±6.16)%、(43.24±7.50)%和(51.06±13.79)%、(47.94±5.36)%,差异有统计学意义(P0.05)。同时TGF-β1 mRNA和蛋白的表达率分别占空白组和pAKD-NC组的(43.72±0.76)%、(44.75±3.78)%和(45.35±2.03)%、(47.71±3.32)%(P0.05),CTGF mRNA和蛋白的表达分别是空白组和pAKD-NC组的(39.32±7.22)%、(39.50±4.58)%和(42.58±5.95)%、(43.74±2.16)%,差异有统计学意义(P0.05)。结论 RAGE特异性siRNA可抑制原代大鼠HSCs RAGE基因的表达,并能有效抑制致纤维化细胞因子的生成。
[Abstract]:Objective the late glycosylation end product receptor, the receptor of advanced glycation end products (RAGEG), can inhibit the expression of RAGE, 伪 -smooth muscle actin alpha-smooth muscle ac-tinin (伪 -SMA) and type I collagen mRNA and protein in hepatic stellate cells in hepatic stellate cells. It plays an important role in the activation of HSCs and the formation of collagen. To investigate the effect of RAGE specific siRNA on the production of fibrogenic cytokines in primary rat HSCs, the expression vector of RAGE specific siRNA was constructed. Primary rat HSCs were isolated and cultured, and the recombinant vector was transfected into primary rat HSCs. The control group was blank group and non-specific siRNA expression vector pAKD-NC group. PCR and Western blot were used to detect the expression of HSCs RAGE- 尾 1 transforming growth factor- 尾 1 (TGF- 尾 1) and connective tissuegrowth factor-CTGF- 尾 1 (CTGF- 尾 1) respectively. Results the HSCs RAGE mRNA and relative molecular weight of pAKD-GR126 were transfected with siRNA expression vector pAKD-GR126. The expression of 42000 RAGE protein was 42.32 卤6.16% in blank group and 43.24 卤7.50% in pAKD-NC group, respectively, and 51.06 卤13.7990 卤5.36% in pAKD-NC group, the difference was statistically significant (P 0.05). Meanwhile, the expression rates of TGF- 尾 1 mRNA and protein in blank group and pAKD-NC group were 43.72 卤0.76 卤3.78% and 45.35 卤2.0335 卤2.035%, respectively. The expression rates of CTGF mRNA and protein in pAKD-NC group were 47.71 卤3.0310% and 47.71 卤3.03% respectively in blank group and pAKD-NC group, which were the same as that in control group and pAKD-NC group (43.72 卤3.78% and 45.35 卤2.0335 卤2.03g), respectively. The expression rates of TGF- 尾 _ 1 mRNA and protein were 43.72 卤3.78% and 45.35 卤2.03% in blank group and pAKD-NC group, respectively. 39.32 卤7.22% and 42.58 卤5.95% and 43.74 卤2.16%, respectively. Conclusion RAGE specific siRNA can inhibit the expression of HSCs RAGE gene in primary rat. And can effectively inhibit the production of fibrogenic cytokines.
【作者单位】：东南大学附属中大医院消化内科;
【基金】：江苏省自然科学基金(BK2009284)
【分类号】：R363

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