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血管内皮生长抑制基因对角膜新生血管形成的抑制作用

Inhibition of corneal neovascularization by vascular endothelia growth inhibitor gene.

作者信息

Wang Hong, Wang Bing, Zhang Zhen-Hai

机构信息

Department of Ophthalmology, the Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.

出版信息

Int J Ophthalmol. 2010;3(3):196-9. doi: 10.3980/j.issn.2222-3959.2010.03.03. Epub 2010 Sep 18.

Abstract

AIM

To evaluate the effect of Effectene™ lipofectine mediated plasmids encoding human pcDNA4-vascular endothelia growth inhibitor (pcDNA(4)-VEGI) gene on corneal neovascularization (CNV).

METHODS

Forty New Zealand albino rabbits were sutured by 5-0 silk on the superior cornea to establish the animal model and divided into 4 random group, ten per each group: group A: transfected by pcDNA(4)-VEGI gene mediated by Effectene™ lipofectine transfection, group B: by Plasmid pcDNA(4), group C: by Effectene™, and group D: by normal saline. Length and area of CNV were measured under slit lamp every day after transfection, immunohistochemistry was used to detected the expression of VEGI protein in cornea at 3, 7, 14 and 21 days.

RESULTS

Average occurrence of CNV in the pcDNA(4)-VEGI gene transfected group (group A) was 6.3 days, in plasmid pcDNA(4) control group (group B) was 3.1 days, in Effectene™ lipofectine control group (group C) was 3.2 days, in normal saline control group (group D) was 3.2 days. Differences between groups A and B, C, D were statistically significant (P<0.01), while differences in groups B, C and D were meaningless (P>0.05). Lenth and average area of CNV in each period in group A was meaningful different from that in groups B, C, and D (P<0.01), while differences in group B, C and D were meaningless (P>0.05). Immunohistochemistry result: VEGI positive cells could be seen in epithelium, stroma, endothelium and the cliff of CNV in group A at 3 days after transfection. VEGI cells changed with the decrease of CNV. None positive cells were in the control groups (groups B, C and D) all the time.

CONCLUSION

Effectene™ lipofectine transfection technique can be effectively used in transfecting pcDNA(4)-VEGI gene into rabbit cornea and the lenth and areas of CNV can be inhibited by VEGI gene.

摘要

目的

评估Effectene™脂质体介导的编码人pcDNA4-血管内皮生长抑制剂(pcDNA(4)-VEGI)基因的质粒对角膜新生血管化(CNV)的影响。

方法

40只新西兰白兔在角膜上方用5-0丝线缝合建立动物模型,随机分为4组,每组10只:A组:用Effectene™脂质体转染介导的pcDNA(4)-VEGI基因转染;B组:用质粒pcDNA(4);C组:用Effectene™;D组:用生理盐水。转染后每天在裂隙灯下测量CNV的长度和面积,采用免疫组织化学法检测转染后3、7、14和21天角膜中VEGI蛋白的表达。

结果

pcDNA(4)-VEGI基因转染组(A组)CNV平均出现时间为6.3天,质粒pcDNA(4)对照组(B组)为3.1天,Effectene™脂质体对照组(C组)为3.2天,生理盐水对照组(D组)为3.2天。A组与B、C、D组之间差异有统计学意义(P<0.01),而B、C、D组之间差异无意义(P>0.05)。A组各时期CNV的长度和平均面积与B、C、D组有显著差异(P<0.01),而B、C、D组之间差异无意义(P>0.05)。免疫组织化学结果:转染后3天,A组上皮、基质、内皮及CNV边缘可见VEGI阳性细胞。VEGI细胞随CNV的减少而变化。对照组(B、C、D组)始终未见阳性细胞。

结论

Effectene™脂质体转染技术可有效将pcDNA(4)-VEGI基因转染至兔角膜,VEGI基因可抑制CNV的长度和面积。

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Inhibition of angiogenesis and breast cancer xenograft tumor growth by VEGI, a novel cytokine of the TNF superfamily.
Int J Cancer. 1999 Jul 2;82(1):131-6. doi: 10.1002/(sici)1097-0215(19990702)82:1<131::aid-ijc22>3.0.co;2-o.
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Ocular neovascularization: an epidemiologic review.眼部新生血管形成:一项流行病学综述。
Surv Ophthalmol. 1998 Nov-Dec;43(3):245-69. doi: 10.1016/s0039-6257(98)00035-6.

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