摘要
目的揭示整合素αvβ3及胸苷磷酸化酶(TYMP)蛋白在宫腔粘连(IUA)发展中的作用,探索粒细胞集落刺激因子(G-CSF)改善宫腔粘连大鼠子宫内膜组织容受性的可能机制,为临床治疗IUA提供参考。方法将12只SD雌性大鼠适应性喂养后,随机分为模型组和治疗组,每组6只。于动情期予以造模,两组大鼠均以95%乙醇损伤双侧宫腔,造模成功后第14天开始,模型组予以生理盐水腹腔注射,治疗组予以G-CSF腹腔注射。造模28 d后处死各组大鼠进行取材,经甲醛固定后行HE染色,观察各组大鼠子宫内膜组织形态,行免疫组织化学法观察整合素αvβ3及TYMP在各组宫腔中的表达情况。另取6只大鼠于动情期造模,造模成功后第14天处死,将子宫取出分为2 cm大小一段,将子宫组织置于G-CSF培养液,在处理后的0、30 min、45 min、1 h行Western blot检测整合素αvβ3及TYMP蛋白的表达水平。收集行宫腔镜检查发现宫腔形态正常及宫腔粘连患者的宫腔灌洗液各15组,采用实时荧光定量聚合酶链反应(qRT-PCR)法检测临床标本中整合素αvβ3及TYMP mRNA的相对表达量。结果HE染色发现,治疗组与模型组相比,子宫形态均有不同程度的恢复,内膜厚度均匀增加,间质内腺体和血管数目均增加。免疫组织化学染色结果提示,与模型组相比,整合素αvβ3及TYMP在治疗组的表达均明显增多。与0 h组比较,G-CSF处理30 min[(1.000±0.058)vs.(1.376±0.121)]、45 min[(1.000±0.058)vs.(2.303±0.331)]、1 h[(1.000±0.058)vs.(2.162±0.477)]可使TYMP的表达升高,差异均有统计学意义(t=6.289、8.662、5.405,P均<0.05);处理45 min[(1.000±0.109)vs.(2.323±0.294)]、1 h[(1.000±0.109)vs.(1.362±0.274)]可使整合素αvβ3的表达升高,差异均有统计学意义(t=9.419、2.747,P均<0.05)。与正常宫腔比较,粘连宫腔整合素αvβ3[(1.000±0.191)vs.(0.573±0.182)]和TYMP[(1.000±0.209)vs.(0.528±0.117)]mRNA的表达均明显下降,差异均有统计学意义(t=2.802、3.413,P均<0.05)。结论整合素αvβ3及TYMP蛋白可作为子宫内膜容受性的评价指标。G-CSF治疗宫腔粘连的作用机制可能与上调子宫内膜中整合素αvβ3及TYMP蛋白的表达有关。
Objective To reveal the role of integrin αvβ3 and thymidine phosphorylase granulocyte colony stimulating factor(TYMP)proteins in the development of uterine adhesions,and to explore the possible mechanisms by which granulocyte colony stimulating factor(G-CSF)improves endometrial tissue tolerance in rats with uterine adhesions,providing more ways to treat intrauterine adhesion(IUA)in clinical practice.Methods Twelve SD female rats were randomly divided into 2 groups after adaptive feeding:model group and treatment group,6 rats in each group.The rats in the two groups were molded during the estrous period,and the bilateral uterine cavities were damaged with 95%ethanol,and the model group was given saline intraperitoneal injection and the treatment group was given G-CSF intraperitoneal injection on the 14 th day after successful molding.The endometrial tissue morphology of each group was observed by HE staining after formaldehyde fixation,and the expression of integrin αvβ3 and TYMP in the uterine cavity of each group was observed by immunohistochemistry.Six rats were selected for membrane creation during the active phase,and the rats were executed on day 14 after successful membrane creation.The expression levels of integrin αvβ3 and TYMP protein were detected by Western blot at 0 min,30 min,45 min and 1 h after treatment.A total of 15 groups of uterine lavage fluid from patients with normal uterine cavity morphology and patients with uterine adhesions were collected by hysteroscopy and detected by quantitative real-time fluorescence polymerase chain reaction(qRT-PCR).The relative expressions of integrin αvβ3 and TYMP mRNA in clinical specimens were examined by qRT-PCR.Results HE staining of the rats in both groups revealed that the uterine morphology was restored to different degrees in the treated group compared with the model group,and the thickness of the endometrium was uniformly increased,and the number of glands and blood vessels in the interstitium were increased.Immunohistochemical staining results suggested that the expression of integrin αvβ3 and TYMP were significantly higher in the treated group compared with the model group.Comparison with the 0 h group,G-CSF treatment for 30 min[(1.000±0.058)vs.(1.376±0.121)],45 min[(1.000±0.058)vs.(2.303±0.331)]and 1 h[(1.000±0.058)vs.(2.162±0.477)]increased the expression of TYMP,and the differences were statistically significant(t=6.289,8.662,5.405,all P<0.05);treatment for 45 min[(1.000±0.109)vs.(2.323±0.294)]and 1 h[(1.000±0.109)vs.(1.362±0.274)]increased the expression of integrinαvβ3,and the differences were statistically significant(t=2.802,3.413,all P<0.05).The expression levels of integrinαvβ3[(1.000±0.191)vs.(0.573±0.182)]and TYMP[(1.000±0.209)vs.(0.528±0.117)]mRNA were significantly lower in the adherent uterus than in the normal uterus,and the differences were statistically significant(t=2.802,3.413,all P<0.05).Conclusions Integrin αvβ3 and TYMP protein could be used as indicators of endometrial tolerance.The mechanism of action of G-CSF in the treatment of uterine adhesions might be related to the upregulation of integrin αvβ3 and TYMP protein expression in the endometrium.
作者
唐洁
朱楚超
TANG Jie;ZHU Chuchao(Wuhan University of Science and Technology of Medicine,Wuhan,Hubei 430000,China;Obstetrics and Gynecology of Central Theater General Hospital,Wuhan,Hubei 430070,China)
出处
《热带医学杂志》
CAS
2023年第7期895-899,1039,共6页
Journal of Tropical Medicine
基金
湖北省卫健委青年人才项目(WJ2017H0035)
关键词
宫腔粘连
粒细胞集落刺激因子
胸苷磷酸化酶
整合素ΑVΒ3
Intrauterine adhesion
Granulocyte colony stimulating factor
Thymidine phosphorylase
Integrin αvβ3
