中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2013年
9期
611-615
,共5页
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
RNA,小分子榦擾%糖基化終產物,高級%受體,細胞錶麵%肝%星形細胞%層黏連蛋白%透明質痠%前膠原
RNA,소분자간우%당기화종산물,고급%수체,세포표면%간%성형세포%층점련단백%투명질산%전효원
RNA,small interfering%Glycosylation end products,advanced%Receptors,cell surface%Liver%Astrocytes%Laminin%Hyaluronic acid%Procollagen
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(PⅢNP)]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HSC RAGE、层粘连蛋白、透明质酸、PⅢNP mRNA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HSCRAGE mRNA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)；PⅢNP mRNA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土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HSC RAGE基因的表达,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.
目的 探討晚期糖基化終產物受體(RAGE)特異性小榦擾RNA(siRNA)在肝纖維化中對纖維化標誌物[層粘連蛋白、透明質痠、血清Ⅲ型前膠原(PⅢNP)]生成的影響.方法 構建RAGE特異性siRNA錶達載體pAKD-GR126,分離培養原代大鼠肝星狀細胞(HSC),將重組載體轉染入原代大鼠HSC,以空白組和轉染非特異性siRNA錶達載體pAKD-NC組為對照,分彆用PCR法和Western免疫印跡法檢測各組原代HSC RAGE、層粘連蛋白、透明質痠、PⅢNP mRNA及蛋白的錶達.正態分佈及方差齊性資料採用方差分析(LSD)及SNK法兩兩比較,非正態分佈及方差不齊資料採用非參數秩和檢驗.結果 轉染特異性siRNA錶達載體pAKD-GR126的原代HSCRAGE mRNA和蛋白的錶達率分彆為空白組和pAKD-NC組的(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)；層粘連蛋白mRNA和蛋白的錶達率分彆為空白組和pAKD-NC組的(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)；透明質痠mRNA和蛋白的錶達率分彆為空白組和pAKD-NC組的(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)；PⅢNP mRNA和蛋白的錶達率分彆為空白組和pAKD-NC組的(56.10±4.18)％、(55.15±2.39)％和(54.40±2.79)％、(53.58土6.18)％(F=141.633,49.670；P均＜0.05).結論 RAGE特異性siRNA可抑製原代大鼠HSC RAGE基因的錶達,併能顯著降低纖維化標誌物(層粘連蛋白、透明質痠、PⅢNP)基因及蛋白的錶達.
목적 탐토만기당기화종산물수체(RAGE)특이성소간우RNA(siRNA)재간섬유화중대섬유화표지물[층점련단백、투명질산、혈청Ⅲ형전효원(PⅢNP)]생성적영향.방법 구건RAGE특이성siRNA표체재체pAKD-GR126,분리배양원대대서간성상세포(HSC),장중조재체전염입원대대서HSC,이공백조화전염비특이성siRNA표체재체pAKD-NC조위대조,분별용PCR법화Western면역인적법검측각조원대HSC RAGE、층점련단백、투명질산、PⅢNP mRNA급단백적표체.정태분포급방차제성자료채용방차분석(LSD)급SNK법량량비교,비정태분포급방차불제자료채용비삼수질화검험.결과 전염특이성siRNA표체재체pAKD-GR126적원대HSCRAGE mRNA화단백적표체솔분별위공백조화pAKD-NC조적(42.32±6.16)％、(43.24±7.50)％화(51.06±13.79)％、(47.94±5.36)％(F=7.791,36.513;P균＜0.05)；층점련단백mRNA화단백적표체솔분별위공백조화pAKD-NC조적(41.07±3.13)％、(40.59±5.87)％화(53.89±2.25)％、(52.46±4.68)％(F=225.111,88.039;P균＜0.05)；투명질산mRNA화단백적표체솔분별위공백조화pAKD-NC조적(45.69±0.87)％、(46.08±2.36)％화(54.20±0.56)％、(52.30±3.42)％(F=178.317,180.646;P균＜0.05)；PⅢNP mRNA화단백적표체솔분별위공백조화pAKD-NC조적(56.10±4.18)％、(55.15±2.39)％화(54.40±2.79)％、(53.58토6.18)％(F=141.633,49.670；P균＜0.05).결론 RAGE특이성siRNA가억제원대대서HSC RAGE기인적표체,병능현저강저섬유화표지물(층점련단백、투명질산、PⅢNP)기인급단백적표체.
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 analyze standard 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-transfected 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 pAKDNC 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 andpAKD-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 PⅢ NP 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).Conclusion RAGE specific siRNA could inhibit the expression of RAGE in primary rat HSC and could significantly lower the expression of fibrosis markers laminin,HA and PⅢ NP at mRNA and protein level.
