Zhu Fengcheng, Liu Mubiao, Pan Ying, Wang Xuefeng, Chen Yanying
Department of Obstetrics and Gynecology, Zhujiang Hospital of Southern Medical University, Guangzhou 510282, China.
Department of Obstetrics and Gynecology, Zhujiang Hospital of Southern Medical University, Guangzhou 510282, China. Email:
Zhonghua Fu Chan Ke Za Zhi. 2015 Jan;50(1):48-53.
To observe the therapeutic effect of NF-κB gene short hairpin RNA (shRNA) on endometriosis and identify the function of NF-κB on the maintenance and development of endometriosis in Macaca fascicularis.
The Macaca fascicularis model of endometriosis was developed, which divided into experimental group, negative control group and simple model group. The high specificity adenovirus vector mediated shRNA targeting NF-κB gene and negative control shRNA adenovirus with no-load NF-κB gene were synthesised. The experimental group injected the adenovirus which carried the NF-κB shRNA into the endometriosis lesions under laparoscopy surgery, the negative control group with no-load shRNA adenovirus and the simple models group injected with normal saline. Four weeks later after the injection, an observed operation was performed through laparoscopy and some lesions were collected. The CD34 immunohistochemistry of these lesions were done to detect the microvessel density, then the variation of the microvessel density among each group were observed. The expression of the NF-κB and proliferating cell nuclear antigen (PCNA) were detected through western blot.
First, the Macaca fascicularis model of endometriosis was successful developed, and the experimental group has an evident atrophy in ectopic lesions compared with the previous. The lesions' microvessel density in experimental group decreased evidently compared with the negative control group and simple model group (0.002 0±0.000 3 versus 0.021 9±0.002 6 versus 0.024 5±0.003 3), and the differences was statistically significant (P < 0.01). The expression of PCNA (0.37±0.17 versus 0.57±0.26 versus 0.57±0.28) and NF-κB (0.338±0.174 versus 0.678±0.021 versus 0.645±0.098) in experiment group was lower than the negative control group and simple model group, the differences were statistically significant (all P < 0.01).
Through targeting suppressed the NF-κB gene expression by NF-κB shRNA, we can inhibit the development of endometriosis through reducing the ability of angiogenesis and cell proliferation of ectopic endometrial cells.
观察NF-κB基因短发夹RNA（shRNA）对猕猴子宫内膜异位症的治疗效果，并明确NF-κB在猕猴子宫内膜异位症维持和发展中的作用。
建立猕猴子宫内膜异位症模型，分为实验组、阴性对照组和单纯模型组。合成靶向NF-κB基因的高特异性腺病毒载体介导的shRNA及空载NF-κB基因的阴性对照shRNA腺病毒。实验组在腹腔镜手术下将携带NF-κB shRNA的腺病毒注入子宫内膜异位症病灶，阴性对照组注入空载shRNA腺病毒，单纯模型组注入生理盐水。注射4周后，通过腹腔镜进行观察性手术并采集部分病灶。对这些病灶进行CD34免疫组织化学检测微血管密度，观察各组微血管密度的变化。通过蛋白质免疫印迹法检测NF-κB和增殖细胞核抗原（PCNA）的表达。
首先，成功建立了猕猴子宫内膜异位症模型，与之前相比，实验组异位病灶明显萎缩。实验组病灶微血管密度明显低于阴性对照组和单纯模型组（0.002 0±0.000 3 vs 0.021 9±0.002 6 vs 0.024 5±0.003 3），差异有统计学意义（P < 0.01）。实验组PCNA（0.37±0.17 vs 0.57±0.26 vs 0.57±0.28）和NF-κB（0.338±0.174 vs 0.678±0.021 vs 0.645±0.098）的表达低于阴性对照组和单纯模型组，差异有统计学意义（均P < 0.01）。
通过NF-κB shRNA靶向抑制NF-κB基因表达，可通过降低异位子宫内膜细胞的血管生成能力和细胞增殖能力来抑制子宫内膜异位症的发展。
