NF-κB因子在普萘洛尔治疗裸鼠血管瘤模型过程中的表达

Expression of NF-kappa B in propranolol treatment for infantile hemangioma in nude mice model

目的研究核转录因子-kappa B(nuclear transcription factor-kappa B,NF-κB)在普萘洛尔治疗婴幼儿血管瘤裸鼠模型过程中的表达情况,探讨其作用及机制。方法手术切除增殖期婴幼儿血管瘤,采用组织块移植法,移植于裸鼠皮下,建立婴幼儿血管瘤的裸鼠移植模型。在移植后的第45天,将移植瘤组织存活的50只实验裸鼠随机分到普萘洛尔干预组(灌注普萘洛尔溶液)和生理盐水对照组(灌注生理盐水),每组25只,进行干预,观察移植瘤的生长变化情况。分别在干预前和干预后第7天、第14天、第21天、第28天,用颈椎脱臼法随机处死两组中各5只裸鼠,每次10只,切取血管瘤组织,检测移植瘤体积和形态改变;采用免疫组化检测移植瘤组织中NF-κBp65、IκB-α、Bcl-2的表达情况;采用半定量RT-PCR检测NF-κBp65 mRNA的表达情况。结果普萘洛尔干预组瘤体较干预前逐渐缩小,并有大量纤维脂肪组织。生理盐水对照组瘤体在第7天、第14天时较干预前增大;第21天、第28天时较干预前缩小,色泽变淡,质地变硬。在干预后各个时间点上,普萘洛尔干预组NF-κBp65的表达低于生理盐水对照组,干预前普萘洛尔干预组NF-κBp65平均光密度值为0.12±0.07,干预后第28天为0.02±0.11。干预前普萘洛尔干预组NF-κBp65 mRNA值为0.981±0.479,干预后第28天为0.267±0.115。普萘洛尔干预组IκB-α的表达高于生理盐水对照组,普萘洛尔干预组Bcl-2的表达低于生理盐水对照组,差异均有统计学意义(P<0.05);普萘洛尔干预组NF-κBp65与Bcl-2呈正相关(r=0.901,P<0.01)。在干预后各个时间点上,普萘洛尔干预组瘤体NF-κBp65 mRNA的相对表达低于生理盐水对照组(P<0.05)。结论普萘洛尔可使裸鼠模型血管瘤组织IκB-α表达升高,NF-κBp65和Bcl-2表达降低。普萘洛尔促进裸鼠模型中的血管瘤消退的作用机制,可能与其降低NF-κB表达,从而促进血管瘤内皮细胞凋亡有关。

Objective To explore the expression and mechanism of nuclear transcription factor-kappa B (NF-κB) in propranolol treatment of infantile hemangioma nude murine model. Methods Proliferative infantile hemangioma tissue was transplanted into nude mice for establishing the model of hemangioma.Fifty surviving mice were randomly divided into two groups of propranolol and saline solution (control).At Days 7,14,21 and 28 post-intervention,five nude mice from each group were randomly sacrificed by cervical dislocation.The samples of transplanted tumor were tested for NF-kappa Bp65,IκB-αand Bcl-2 by immunohistochemistry.And the expression of NF-kappa Bp65 mRNA in samples was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Results After intervention,the transplanted tumors in propranolol group shrank,became lighter and hardened.In saline group,transplanted tumors enlarged initially at Days 7 and 14 and there were numerous hemangioma endothelial cells.Then at Days 21 and 28,tumor tissue shrank,became lighter and hardened.At each timepoint,NF-κBp65 in propranolol group was lower than that in saline group ( P <0.05);At various timepoints post-intervention,the expression of NF-kappa Bp65 was lower in ropranolol group than that in normal saline group.The average optical density of NF-kappa Bp65 was 0.12±0.07 pre-intervention and 0.02±0.11 at Day 28 post-intervention.The value of NF-kappa 65 mRNA in propranolol group was 0.981±0.479 pre-intervention and 0.267±0.115 at Day 28 post-intervention.IκB-αwas higher in propranolol group than that in saline group ( P <0.05);Bcl-2 was lower in propranolol group than that in saline group ( P <0.05).At the same time,a positive linear correlation existed between NF-kappa Bp65 and Bcl-2 in propranolol group ( r =0.901, P <0.01).During each phase,NF-kappaBp65 mRNA was significantly lower in propranolol group than that in saline group ( P <0.05). Conclusion Propranolol may up-regulate the expression of kappa b-alpha and down-regulate the expressions of NF-kappa Bp65 and bcl-2 in hemangioma tissues of nude mice.The mechanism by which propranolol promotes the regress of hemangioma in nude murine model may be related to its effect of suppressing the anti-apoptosis mediated by NF-kappa B.