晚期糖基化终产物受体特异性小干扰RNA抑制肝星状细胞表达初探  

Expression of hepatic stellate cells inhibited by receptor for advance glycoxidation end products specific small interfering

作　　者：刘雅静[1] 夏金荣[1] 

机构地区：[1]东南大学附属中大医院消化内科,南京210009

出　　处：《中华消化杂志》2013年第9期611-615,共5页Chinese Journal of Digestion

基　　金：江苏省自然科学基金（BK2009284）

摘　　要：目的探讨晚期糖基化终产物受体（RAGE）特异性小干扰RNA（siRNA）在肝纤维化中对纤维化标志物[层粘连蛋白、透明质酸、血清Ⅲ型前胶原（PHINP）]生成的影响。方法构建RAGE特异性siRNA表达载体pAKD—GR126，分离培养原代大鼠肝星状细胞（HSC），将重组载体转染入原代大鼠HSC，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免疫印迹法检测各组原代HSCRAGE、层粘连蛋白、透明质酸、PⅢNPmRNA及蛋白的表达。正态分布及方差齐性资料采用方差分析（LSD）及SNK法两两比较，非正态分布及方差不齐资料采用非参数秩和检验。结果转染特异性siRNA表达载体pAKD-GR126的原代HSCRAGEmRNA和蛋白的表达率分别为空白组和pAKD-NC组的（42．32±6．16）％、（43．24±7．50）％和（51．06±13．79）％、（47．94±5．36）％（F=7．791，36．513；P均〈0．05）；层粘连蛋白mRNA和蛋白的表达率分别为空白组和pAKD-NC组的（41．07±3．13）％、（40．59±5．87）％和（53．89±2．25）％、（52．46±4．68）％（F=225．111，88．039；P均〈0．05）；透明质酸mRNA和蛋白的表达率分别为空白组和pAKD-NC组的（45．69±0．87）％、（46．08±2．36）％和（54．20±0．56）％、（52．30±3．42）％（F=178．317，180．646；P均〈0．05）；PⅢNPmRNA和蛋白的表达率分别为空白组和pAKD-NC组的（56．10±4．18）％、（55．15±2．39）％和（54．40±2．79）％、（53．58±6．18）％（F=141．633，49．670；P均〈0．05）。结论RAGE特异性siRNA可抑制原代大鼠HSCRAGE基因的表扶，并能显著降低纤维化标志物（层粘连蛋白、透明质酸、PⅢNP）基因及蛋白的表达。Objective To investigate the effect of specific small interfering RNA （siRNA） targeting receptor of advanced glycation end products （RAGE） on the production of fibrosis markers （laminin, hyaluronic acid （HA） and N-terminal procollagen Ⅲ propeptide （PⅢ NP） in hepatic fibrosis （HF）. Methods The expression vectors of specific siRNA targeting RAGE were constructed. Primary rat hepatic stellar cells （HSC） were isolated and cultured. The primary rat HSC were transfected with the recombinant vector. The blank control group and unspecific siRNA vector pAKD- NC transfected group were as controls. The expressions of RAGE,laminin, HA and PⅢ NP at mRNA and protein levels were detected by real time polymerase chain reaction and Western blot, respectively. Least-significant difference （LSD） and Student-Newman-Keuls （SNK） were performed to analyzestandard normal distribution or homogeneous variance. Non normal distribution and heterogeneity of variance data were analyzed by non parametric Wilcoxon test. Results The expressions of RAGE at mRNA and protein levels in pAKD-GR126-transfeeted primary HSC were （42. 32±6. 16 ）%, （43.24±7.50）%, （51.06±13.79）% and （47.94±5.36）% in blank control group and pAKD-NC group （F=7. 791 and 36. 513,all P〈0.05）. The expressions of laminin at mRNA and protein levels were （41.07±3.13） %, （40.59±5.87） %, （53.89±2.25） % and （52.46±4.68） % in blank control group and pAKD-NC group （F=225. 111 and 88. 039, all P〈0. 05）. The expressions of HA at mRNA and protein levels were （45. 69±0. 87）%, （46. 08±2. 36）%, （54. 20±0. 56）% and （52.30±3.42）% in blank control group and pAKD-NC group （F 178. 317 and 180. 646,all P〈 0.05）. The expressions of P ⅢNP mRNA at mRNA and protein levels were （56. 10 ±4. 18）%, （55.15±2.39）%, （54. 40±2. 79）% and （53. 58±6. 18）% in blank control group and pAKD-NC group （F=141. 633 and 49. 670,all P〈0.05）. Conelttsion RAGE specific siRNA could

关 键 词：RNA 小分子干扰 糖基化终产物 高级 受体 细胞表面 肝 星形细胞 层黏连 蛋白 透明质酸 前胶原 

分 类 号：R575.2[医药卫生—消化系统]