周细胞在子宫内膜异位症血管生成中作用的研究

发布时间：2018-06-02 16:36

本文选题：子宫内膜异位症 + 血管生成　；参考：《天津医科大学》2014年硕士论文

【摘要】：目的：本实验通过测定子宫内膜异位症(EMs, endometriosis)患者异位、在位、正常内膜及不同时相的裸鼠异位病灶中周细胞数目(PN, Pericytes number)、微血管密度(MVD, microvessel density)以及PN/MVD：北值,研究探讨周细胞在EMs血管生成中的作用。方法：选取EMs患者异位内膜20例(异位组)、在位内膜40例(增殖期、分泌期各20例)(在位组)及非子宫内膜异位症患者子宫内膜40例(增殖期、分泌期各20例)(对照组)。利用异体移植方法建立40只裸鼠人EMs模型,并随机平均分为A组、B组、C组、D组,分别于建模后7天、14天、21天、28天时处死并获取异位病灶。观察裸鼠异位病灶肉眼及镜下组织学形态,并采用免疫组化法检测三组人子宫内膜及四组裸鼠异位病灶中PN、MVD的表达,并对二者的比值(PN/MVD)进行比较。结果：(1)裸鼠EMs模型成功率达100%；(2)人异位内膜MVD值高于正常内膜和在位内膜(P0.05),在位内膜无论增殖期还是分泌期与相应的正常内膜相比差异均无统计学意义,在位内膜及正常内膜组内比较均分泌期高于增殖期(P0.05)；(3)人异位内膜PN值低于正常内膜和在位内膜(P0.05),在位内膜无论增殖期还是分泌期与相应的正常内膜相比差异均无统计学意义,在位内膜及正常内膜组内比较均增殖期高于分泌期(P0.05)；(4)人异位内膜PN/MVD值低于正常内膜和在位内膜,在位内膜无论增殖期还是分泌期均低于相应的正常内膜(P0.05),在位内膜及正常内膜组内比较均增殖期高于分泌期(P0.05)；(5)A组裸鼠开腹探查见异位病灶呈清亮水疱状,镜下可见点簇状新生血管形成,MVD=35.70±3.16, B组裸鼠开腹探查见异位病灶呈红色囊泡状,表面可见血管走形,镜下可见腺体增生,间质中血管生成明显,MVD=68.60±9.07, C组裸鼠开腹探查见异位病灶呈暗红色囊状或结节状,欠新鲜,镜下可见腺体丰富、呈增殖期子宫内膜表现,间质新生血管数目减少、血管壁结构完整,MVD=53.50±3.57, D组裸鼠开腹探查见异位病灶呈黄白色囊状或结节状、质稍硬,镜下可见腺体数目减少,部分腺体及间质纤维化,间质新生血管数目进一步减少,可见部分血管结构破坏；MVD=48.00+4.99；四组裸鼠异位病灶MVD值差异具有统计学意义(P0.05)；(6)A-D组裸鼠异位病灶中PN与MVD的变化趋势平行,分别为(27.30+2.50)、(79.40±10.68)、(64.70±4.64)、(38.30±4.50),差异均有统计学意义(P0.05)；(7)A-C组裸鼠PN/MVD值呈上升趋势,在C组裸鼠异位病灶中达到峰值,D组裸鼠异位病灶中PN/MVD值同C组比较呈下降趋势,但仍高于A组裸鼠,差异有统计学意义(P0.05)。结论：(1)采用异体移植方法可在裸鼠体内较完整地复制人类子宫内膜异位症,可实现对EMs发展过程的动态观察及相关实验研究,方法可行、成功率高,所得结果具有参考价值；(2)四组裸鼠异位病灶中PN值随着病灶中血管的生长、消退发生相应的改变,与相应内膜中MVD值变化趋势相一致,提示周细胞在EMs血管生成中起着重要作用；(3)EMs患者异位内膜中PN/MVD值低于在位和正常内膜,C组裸鼠异位病灶中PN/MVD值达峰值,结合肉眼与镜下所见,提示EMs病灶中周细胞覆盖率与血管稳定性高度一致性；(4)EMs患者在位内膜无论增殖期还是分泌期中的PN/MVD值均低于相应的正常内膜,进一步佐证了“在位内膜决定论”,在位子宫内膜中周细胞覆盖率低可能是EMs发病的因素之一；(5)周细胞在EMs血管生成中的作用机制还有待进一步研究。
[Abstract]:Objective: to determine the role of pericytes in EMs angiogenesis by measuring the number of pericytes (PN, Pericytes number), microvascular density (MVD, microvessel density) and PN/ MVD: the North value of ectopic and normal endometriosis (EMs, endometriosis) patients with heterotopic endometriosis, normal endometrium and different phase of nude mice.
Methods: 20 ectopic endometrium (ectopic endometrium) of EMs patients (ectopic endometrium), 40 cases of eutopic endometrium (proliferating, 20 secretory phase) and 40 non Endometriosis Endometrium endometrium (20 cases in the proliferation period and 20 secretory phase) were selected (control group). The EMs model of 40 nude mice was established by allograft transplantation, and was randomly divided into A group, B group, C group and D group. The expression of PN, MVD, and the expression of PN, MVD in the endometrium of three groups of human endometrium and four groups of nude mice were detected by immunohistochemical method, and the ratio of two (PN/MVD) was compared between the three groups of endometrium and four groups of nude mice.
Results: (1) the success rate of EMs model in nude mice was 100%. (2) the MVD value of ectopic endometrium was higher than normal endometrium and eutopic endometrium (P0.05). There was no statistical difference between the eutopic endometrium and the normal endometrium in the eutopic endometrium and the secretory phase. The secretory phase of the eutopic endometrium and the normal endometrium was higher than that in the proliferation period (P0.05); (3) the human ectopic endometrium was heterotopic. The PN value of the intima was lower than that of the normal intima and the eutopic endometrium (P0.05). There was no significant difference between the eutopic and secretory phase in the eutopic endometrium and the normal endometrium, in the eutopic endometrium and in the normal endometrium, the proliferation period was higher than that of the secretory phase (P0.05); (4) the PN/MVD value of the human endometrium was lower than that of the normal intima and the eutopic endometrium. The proliferation and secretory phase of the membrane were lower than that of the corresponding normal endometrium (P0.05), and the proliferation period in the eutopic endometrium and the normal endometrium was higher than that in the secretory phase (P0.05). (5) the open blistering of the A group in nude mice showed that the ectopic lesion showed a clear blister shape, the cluster shaped neovascularization was found under the microscope, MVD=35.70 + 3.16, and the open abdominal exploration of nude mice in group B was found to be ectopic disease. The focus was red vesicles, the surface of the blood vessel was visible, the glandular hyperplasia was seen under the microscope, the angiogenesis in the interstitium was obvious, MVD=68.60 + 9.07. C group nude mice open abdominal exploration showed that the ectopic lesion was dark red saclike or nodular, it was not fresh, the gland was abundant in the microscope, showed the endometrium in the proliferation stage, the number of interstitial neovascularization decreased and the vascular wall structure was reduced. Complete, MVD=53.50 + 3.57, D group nude mice open abdominal exploration showed that the ectopic lesion was yellow white cystic or nodular, slightly hard, the number of glandular decreased, some gland and interstitial fibrosis, the number of interstitial neovascularization was further reduced, and some vascular structure destruction; MVD= 48.00+4.99; the difference of MVD value of four groups of nude mice with heterotopic focus had unification. Study significance (P0.05); (6) the change trend of PN and MVD in the nude mice of A-D group was parallel, (27.30+2.50), (79.40 + 10.68), (64.70 + 4.64), (38.30 + 4.50), and the difference was statistically significant (P0.05). (7) the PN/MVD value of nude mice in the A-C group was up, reached the peak in the heterotopic nude mice of the C group, and the PN/MVD value of the nude mice in the D group was in the nude mice. Compared with group C, there was a downward trend, but it was still higher than that in group A, and the difference was statistically significant (P0.05).
Conclusions: (1) human endometriosis can be replicated completely in nude mice by allograft. The dynamic observation and related experimental study of EMs development process can be realized. The method is feasible, the success rate is high, and the results have reference value. (2) the value of PN in four groups of nude mice heterotopic focus decreases with the growth of the blood vessels in the lesion. The corresponding changes were consistent with the changes in the MVD values in the corresponding endometrium, suggesting that the pericytes play an important role in the angiogenesis of EMs; (3) the PN/MVD value in the ectopic endometrium of the EMs patients is lower than that in the eutopic and normal endometrium, and the peak value of the PN/MVD value in the ectopic lesion of the C group nude mice, combining with the naked eye and the mirror, suggests the coverage rate of the pericytes in the EMs focus. The high consistency of vascular stability; (4) the PN/MVD values in the eutopic endometrium of EMs patients are lower than that of the corresponding normal endometrium in both proliferative and secretory phase, which further confirms the "eutopic endometrial determinism". The low pericyte coverage rate in the eutopic endometrium may be one of the factors in the pathogenesis of EMs; (5) the role of pericytes in the angiogenesis of EMs. The mechanism remains to be further studied.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R711.71

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