Small hairpin RNA targeting inhibition of NF-κB gene in endometriosis therapy of Macaca fascicularis / 中华妇产科杂志

Objective 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.Methods 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.Results 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).Conclusion 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.

Adenovirus vector-mediated short hairpin RNA targeting nuclear factor-κB suppresses proliferation of endometrial cells of Macaca fascicularis in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the effect of a high specific adenovirus vector-mediated shRNA targeting nuclear factor-κB (NF-κB) on cell proliferation of the endometrium of Macaca fascicularis.</p><p><b>METHODS</b>The adenoviral vector NF-κB-p65-shRNA and the empty vector were separately trasnfected in cultured endometrial cells of Macaca fascicularis. The changes in the expression of the target gene protein and apoptotic proteins, cell proliferation, and cell cycle distribution were observed after the transfection.</p><p><b>RESULTS</b>Compared with the control cells, infection of the endometrial cells with the NF-κB-p65-shRNA adenovirus significantly increased the expression levels of apoptotic proteins, promoted apoptosis of the endometrial cells, and reduced the cells in division?stage.</p><p><b>CONCLUSIONS</b>NF-κB-p65 shRNA adenovirus can effectively promote apoptosis of endometrial cells and inhibit the proliferation of endometrial cells of Macaca fascicularis.</p>

Effect of interleukin-1β on expressions of activin A and its related factors in cultured endometrial stromal cells from patients with endometriosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of interleukin-1β (IL-1β) on the expressions activin A, follistatin, and cripto in cultured human endometrial stromal cells (HESCs) form patients with endometriosis.</p><p><b>METHODS</b>Cultured HESCs were stimulated with 250, 500, and 750pg/ml IL-1β, and the mRNA and protein expressions of activin A, follistatin, and cripto were assayed using real-time reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>IL-1β treatment caused significant dose-dependent increments of the mRNA and protein expressions of activin A and follistatin and of the mRNA expression of cripto in cultured HESCs.</p><p><b>CONCLUSION</b>IL-1β can affect the expressions of activin A, follistatin and cripto in HESCs from patients with endometriosis.</p>