中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2009年
5期
360-364
,共5页
陈木开%刘隽华%马春光%韩建德
陳木開%劉雋華%馬春光%韓建德
진목개%류준화%마춘광%한건덕
RNA干扰%RNA%小分子干扰%单纯疱疹病毒2型%基因表达%疱疹%生殖器
RNA榦擾%RNA%小分子榦擾%單純皰疹病毒2型%基因錶達%皰疹%生殖器
RNA간우%RNA%소분자간우%단순포진병독2형%기인표체%포진%생식기
RNA interference%RNA%small interfering%Herpes simplex virus 2%Gene expression%Herpes genitalis
目的 探讨联合靶向特异性小干扰RNA(siRNA)对疱疹病毒(HSV)2的体外特异抑制作用.方法 将体外合成的靶向HSV-2编码包膜糖蛋白gB的UL27.2基因、靶向DNA结合蛋白UL29.2基因的siRNA和该两种联合靶向特异性siRNA制剂,共转染Vero细胞并感染HSV-2,观察靶基因的表达情况、Vero细胞病变、空斑减数实验和子代病毒滴度并进行各组间比较:结果特异性UL27.2siRNA、UL29.2 siRNA和联合siRNA转染Vero细胞后,均能不同程度抑制各HSV-2临床株感染所导致的细胞病变,对病毒增殖抑制率分别为63.9%、86.7%和93.3%.实时定量PCR检测各组siRNA分别对UL27.2和UL29.2两个基因的表达,结果显示UL27.2 siRNA和UL29.2 siRNA对各自的靶向基因均有抑制,而联合靶向siRNA制剂对两个基因的抑制率最高.结论 联合靶向特异性siRNA制剂能有效抑制HSV-2的感染和复制.
目的 探討聯閤靶嚮特異性小榦擾RNA(siRNA)對皰疹病毒(HSV)2的體外特異抑製作用.方法 將體外閤成的靶嚮HSV-2編碼包膜糖蛋白gB的UL27.2基因、靶嚮DNA結閤蛋白UL29.2基因的siRNA和該兩種聯閤靶嚮特異性siRNA製劑,共轉染Vero細胞併感染HSV-2,觀察靶基因的錶達情況、Vero細胞病變、空斑減數實驗和子代病毒滴度併進行各組間比較:結果特異性UL27.2siRNA、UL29.2 siRNA和聯閤siRNA轉染Vero細胞後,均能不同程度抑製各HSV-2臨床株感染所導緻的細胞病變,對病毒增殖抑製率分彆為63.9%、86.7%和93.3%.實時定量PCR檢測各組siRNA分彆對UL27.2和UL29.2兩箇基因的錶達,結果顯示UL27.2 siRNA和UL29.2 siRNA對各自的靶嚮基因均有抑製,而聯閤靶嚮siRNA製劑對兩箇基因的抑製率最高.結論 聯閤靶嚮特異性siRNA製劑能有效抑製HSV-2的感染和複製.
목적 탐토연합파향특이성소간우RNA(siRNA)대포진병독(HSV)2적체외특이억제작용.방법 장체외합성적파향HSV-2편마포막당단백gB적UL27.2기인、파향DNA결합단백UL29.2기인적siRNA화해량충연합파향특이성siRNA제제,공전염Vero세포병감염HSV-2,관찰파기인적표체정황、Vero세포병변、공반감수실험화자대병독적도병진행각조간비교:결과특이성UL27.2siRNA、UL29.2 siRNA화연합siRNA전염Vero세포후,균능불동정도억제각HSV-2림상주감염소도치적세포병변,대병독증식억제솔분별위63.9%、86.7%화93.3%.실시정량PCR검측각조siRNA분별대UL27.2화UL29.2량개기인적표체,결과현시UL27.2 siRNA화UL29.2 siRNA대각자적파향기인균유억제,이연합파향siRNA제제대량개기인적억제솔최고.결론 연합파향특이성siRNA제제능유효억제HSV-2적감염화복제.
Objective To investigate the inhibition of herpes simplex virus (HSV)2 by combined targeting small interfering RNAs(siRNAs) in vitro. Methods Two different siRNAs,which target HSV-2 UL27.2 gene encoding envelope glycoprotein gB and UL29.2 gene encoding DNA binding protein, were synthesized chemically in vitro. Combined siRNAs composed of these two target-specific siRNAs were transcripted and transfected into Vero cells to infect HSV-2. Real-time PCR was used to measure the effect of siRNA on target gene expression, and estimate the anti-HSV-2 effect with pathological changes, plaque reduce test and the concentrations of filial generation virus. The results were compared among groups. Results Specific U L27.2 siRNA, UL29.2 siRNA and the combined siRNA transfected into Veto cells could inhibit the cytopathic effect due to clinical strains of HSV-2 infection in varying degrees, siRNAs could effectively inhibit virus growth by 63.9%, 86.7% and 93.3%, respectively. Real-time PCR detection of the UL27.2 and UL29.2 gene expression showed that UL27.2 siRNA and UL29.2 siRNA inhibited the expression of the respective targeting genes, while combined siRNA showed the highest inhibition rate of expression. Conclusion Combined target-specific siRNAs may obviously inhibit the replication and infection of HSV-2.
