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学科主题: 妇产科学
题名:
1 POP阴道壁组织基质金属蛋白酶与胶原的表达及意义 2 P物质在内异症慢性盆腔痛子宫骶韧带中的表达及意义
作者: 胡艳
答辩日期: 2016-05-12
导师: 吴瑞芳
专业: 妇产科学
授予单位: 北京大学
授予地点: 北京大学深圳医院
学位: 博士
关键词: 盆腔脏器脱垂 ; 基质金属蛋白酶-8 ; 基质金属蛋白酶-1 ; 组织基质金属蛋白酶抑制物-1 ; 胶原蛋白 ; P物质 ; 慢性盆腔痛 ; 子宫内膜异位症
其他题名: 1 Expressions and significance of metalloproteinase and collagen in vaginal wall tissues of patients with pelvic organ prolapsed
分类号: R711.5
摘要:

一、

研究目的: 探讨基质金属蛋白酶-8(matrix metalloproteinase-8,MMP-8)、基质金属蛋白酶-1(matrix metalloproteinase-1,MMP-1)、组织基质金属蛋白酶抑制物(tissue inhibitor of metalloproteinase-1, TIMP-1)及I型、Ⅲ型胶原蛋白与女性盆腔脏器脱垂的关系。

研究方法 : 选择2012年10月至2014年12月在我院因盆腔脏器脱垂(pelvic organ prolapse, POP)行盆底手术的60例患者作为研究对象,60例患者均伴有Ⅱ度或以上阴道前壁脱垂,其中阴道前壁Ⅱ度脱垂者23例、为Ⅱ度脱垂组,Ⅲ-Ⅳ度脱垂者37例、为Ⅲ-Ⅳ度脱垂组;以同期因妇科良性疾病住院手术无POP的患者35例为对照组。手术时取POP患者脱垂的阴道前壁组织,对照组取尿道中段阴道前壁组织;应用链霉素抗生物素蛋白-过氧化物酶连接(streptavidin-perosidase,SP)免疫组化法染色检测其MMP-8、MMP-1、TIMP-1及Ⅰ型、Ⅲ型胶原蛋白的相对表达量;应用实时荧光定量PCR法检测其MMP-8、MMP-1、TIMP-1及Ⅰ型、Ⅲ型前胶原蛋白mRNA的相对表达量。

 

1

结果:1.SP免疫组化法染色检测MMP-8、MMP-1、TIMP-1及Ⅰ型、Ⅲ型胶原蛋白在Ⅱ度脱垂组、Ⅲ-Ⅳ度脱垂组和对照组阴道前壁组织中均有表达,MMP-8、MMP-1、TIMP-1的阳性表达为棕黄色颗粒,其中MMP-1阳性颗粒表达位于细胞浆内及细胞膜上,MMP-8、TIMP-1阳性颗粒表达位于细胞浆内;Ⅰ型、Ⅲ型胶原蛋白免疫组化染色阳性表达为棕色的条索状或束状结构,主要表达于细胞间质中。2. 以SP免疫组化法检测阴道前壁中MMP-8、MMP-1、TIMP-1及Ⅰ型、Ⅲ型胶原蛋白的阳性染色面积百分比(%)平均值,在对照组分别为:6.8363±1.2763、7.7940±0.5506、25.6723±1.1563、55.5343±5.6148、26.9800±1.5080;在Ⅱ度脱垂组分别为:12.2065±1.7220、8.5530±1.1405、24.4287±2.0149、40.0478±4.7836、26.3087±2.0181;在Ⅲ-Ⅳ度脱垂组分别为:13.9703±2.3672、14.4797±2.3603、20.8911±3.7175、30.1216±4.6210、23.8622±1.4754。其中MMP-8、MMP-1蛋白在Ⅲ-Ⅳ度脱垂组阴道前壁中的表达量明显高于Ⅱ度脱垂组和对照组,MMP-8蛋白在Ⅱ度脱垂组阴道前壁中的表达量明显高于对照组,差异均具有统计学意义(P<0.05);TIMP-1和Ⅰ型、Ⅲ型胶原蛋白在Ⅲ-Ⅳ度脱垂组阴道前壁中的表达量明显低于Ⅱ度脱垂和对照组,Ⅰ型胶原蛋白在Ⅱ度脱垂组阴道前壁中的表达量明显低于对照组,差异均具有统计学意义(P<0.05)。3. Ⅰ/Ⅲ型胶原蛋白比值在对照组、Ⅱ度脱垂组及Ⅲ-Ⅳ度脱垂组分别为:2.0643±0.2372、1.5276±0.1878和1.2686±0.2186,三组之间的差异均具有统计学意义(P<0.05)。阴道前壁中Ⅰ、Ⅲ型胶原蛋白表达量与MMP-8、MMP-1蛋白表达量呈负相关,与TIMP-1蛋白表达量呈正相关(P<0.01)。4. 实时荧光定量PCR法检测阴道前壁组织中MMP-8、MMP-1、TIMP-1及Ⅰ型、Ⅲ型前胶原蛋白mRNA的相对表达量在对照组分别为:0.0264±0.0079 、0.0325±0.0042、0.5401±0.0556、0.6429±0.0445、0.4494±0.0514;在Ⅱ度脱垂组分别为:0.0616±0.0096、0.0327±0.0077、0.5139±0.0480、0.6382±0.0534、0.4419±0.0567;在Ⅲ-Ⅳ度脱垂组分别为:0.0801±0.0081、0.0705±0.0067、0.3944±0.5420、0.6221±0.0402、0.4351±0.0313。其中MMP-8、MMP-1mRNA在Ⅲ-Ⅳ度脱垂组阴道前壁中的表达量明显高于Ⅱ度脱垂组和对照组,MMP-8mRNA在Ⅱ度脱垂组阴道前壁中的表达量明显高于对照组,差异均具有统计学意义(P<0.05)。TIMP-1mRNA在Ⅲ-Ⅳ度脱垂组阴道前壁中的表达量明显低于Ⅱ度脱垂组和对照组,差异具有统计学意义(P<0.05)。而Ⅰ型、Ⅲ型前胶原蛋白 mRNA在三组阴道前壁组织中的表达量无明显差异(P>0.05)。

结 论 :1.盆腔脏器脱垂患者阴道前壁组织中Ⅰ、Ⅲ型胶原蛋白含量减少、Ⅰ/Ⅲ型胶原蛋白比值降低,与POP病变相关;而Ⅰ型胶原蛋白含量减少、Ⅰ/Ⅲ型胶原蛋白比值下降与病变严重程度直接相关。2.Ⅰ、Ⅲ型胶原蛋白含量减少与局部MMP-8、MMP-1表达增强,TIMP-1表达下降密切相关;MMP-8、MMP-1在盆腔脏器脱垂患者阴道前壁组织中的表达量明显增加;而Ⅰ型、Ⅲ型前胶原蛋白 mRNA在POP患者与对照组中的表达量无明显差异,提示局部组织中基质金属蛋白酶表达增加导致胶原蛋白含量减少及组成改变,在盆腔脏器脱垂发生发展中起重要作用。

二、

研究目的:研究P物质在子宫内膜异位症所致慢性盆腔痛患者子宫骶韧带中的表达,探讨局部P物质改变与慢性盆腔痛的关系。

研究方法:选择2013年6月~2014年12月26例因子宫内膜异位症所致慢性盆腔疼痛施行手术治疗的26例患者为研究对象,同期因妇科良性疾病行子宫切除术非子宫内膜异位症及慢性盆腔痛的患者33例为对照。术中取患者子宫骶韧带组织,应用实时荧光定量PCR法及免疫组化链霉素抗生物素蛋白-过氧化物酶连接法(streptavidin peroxidase conjugated method,SP)法检测P物质mRNA和蛋白在两组患者子宫骶韧带中的相对表达水平。

结果:1.研究组和对照组子宫骶韧带中均有P物质蛋白及mRNA表达。2.研究组子宫骶韧带中P物质mRNA的相对表达量中位数位0.2129、四分位数间距为0.6954,P物质蛋白的阳性染色面积百分比(%)为25.6531±7.0761;对照组分别为0.0364、0.1674和20.2915±4.3665。研究组子宫骶韧带中P物质mRNA的相对表达量和蛋白的阳性染色面积百分比均明显高于对照组(Z=2.141,P=0.032;t=3.614,P=0.001)。

结论 :子宫内膜异位症所致慢性盆腔痛患者子宫骶韧带中P物质mRNA的相对表达量和蛋白的阳性染色面积百分比均较非子宫内膜异位症无慢性盆腔痛患者为高,提示子宫骶韧带中P物质表达增加可能是子宫内膜异位症引起慢性盆腔痛的原因。

英文摘要:

一、

Objective:To investigate the relationship between MMP-8、MMP-1、TIMP-1、type I and type III collagen and female pelvic organ prolapse.

Methods:60 cases of patients with pelvic organ prolapse were treated with pelvic surgery as the research objects, according to the prolapse degree of anterior vaginal wall, the patients were divided into two groups: POP II group ( 23 cases ) and POP III-IV group (37 cases). 35 patients without POP  as control group.   Take the tissues in the anterior vaginal wall of prolapsed of the 60 totally patients during operation as specimen, in control group, the anterior wall of vagina was ed. Streptavidin-Perosidase immunohistochemistry technique was used to evaluate the protein expressions of MMP-8、MMP-1、TIMP-1、type I and type III collagen in the specimen.  Real-time quantitative reverse tranion-polymerase chain reaction (RT-PCR)was used to detect the mRNA expressions of MMP-8、MMP-1、 TIMP-1、 type I and type III pro-collagen in the specimen.

 

3

Results:1. The protein expressions of MMP-8、MMP-1、TIMP-1、type I and type III collagen were  all observed in anterior vaginal wall in both the POP II group 、 POP III-IV group and the control group; the positive expressions of MMP-8、MMP-1、TIMP-1 showed brown-yellow granules among them, MMP-1 expression in the cytoplasm and cell membrane, MMP-8 and TIMP-1 expressions in the cytoplasm. Immunohistochemistry staining results showed that the positive expressions of type I and type III  collagen were brown streak or sarciniform structure which mainly occurred in the intercellular substance. 2. Average relatively positive areas of MMP-8、MMP-1、TIMP-1、type I and type III collagen detected by the Streptavidin-Perosidase immunohistochemistry technique in anterior vaginal wall in control group  were 6.8363±1.2763 、7.7940±0.5506、25.6723±1.1563、55.5343±5.6148、26.9800±1.5080; POP II group were 12.2065±1.7220、8.5530±1.1405、24.4287±2.0149 、40.0478±4.7836 、26.3087±2.0181;POP III - IV group were 13.9703±2.3672、14.4797±2.3603 、20.8911±3.7175、30.1216±4.6210、23.8622±1.4754. The MMP-8 and MMP-1 positive  areas  were significantly higher in POP III-IV group than that in POP II and control group, The MMP-8 positive   area  was  significantly higher in POP II group than that in control group(P<0.05); while the TIMP-1、 type I and type III collagen positive   areas  were significantly lower in POP III -IV group than that in POP II and control group(P<0.05), The Type I collagen positive   area was  significantly lower in POP II group than that in control group(P<0.05). 3. Type I and III collagen ratio in the control group, POP II   group and POP  group respectively: 2.0643 ± 0.2372, 1.5276 ± 0.1878 and 1.2686 ± 0.2186, differences among the three groups were statistically significant(P<0.05). the expressions of type I and type III collagen  decreased with increased expressions of MMP-8 and MMP-1  and decreased expression of TIMP-1 are both significantly linear correlation. 4. The mRNA expressions of MMP-8、MMP-1、TIMP-1、type I and type III pro-collagen in anterior vaginal wall with control group  were  0.0264±0.0079、0.0325±0.0042、0.5401±0.0556、0.6429±0.0445、0.4494±0.0514;POP II group were 0.0616±0.0096、0.0327±0.0077、0.5139±0.0480、0.6382±0.0534、0.4419±0.0567;POP III-IV group were 0.0801±0.0081、0.0705±0.0067、0.3944±0.5420、0.6221±0.0402、0.4351±0.0313. The mRNA expressions of MMP-8 and MMP-1 were significantly higher in POP III-IV group than that in POP II and control group, the mRNA expression of MMP-8 was significantly higher in POP II group than that in control group(P<0.05),while the mRNA expression of TIMP-1 was significantly lower in POP III-IV group than that in POP II and control group(P<0.05). The mRNA expressions of type I and type III pro- collagen among the three groups  were no significant difference(P>0.05). Conclusions:1. The decrease in expressions of type I and III collagen and I/III collagen ratio in anterior vaginal wall of POP patients is directly related to the prolapse degree of anterior vaginal wall.2. The decrease of I、III collagen    is closely related to the local increased expressions of MMP-8、MMP-1 and decreased expression of TIMP-1; The results suggest that the increase  of metalloproteinases are directly related to the decrease of the contents of collagen and the change of the composition in local tissue which play a very important role in the pathogenesis of POP.

二、

Objective:To investigate the relationship between substance P expression in uterosacral ligament and onic pelvic pain caused by endometriosis.  Methods  26 cases of patients with onic pelvic pain caused by endometriosis were treated with surgery as the research group,  33 patients without onic pelvic pain and endometriosis as control group. Streptavidin  Perosidase immunohistochemistry technique was used to evaluate the expression of substance P in uterosacral ligament. Real-time quantitative reverse tranion  polymerase chain reaction  was used to detect the mRNA expression of substance P in uterosacral ligament. Results  The median of mRNA relative expression of substance P was  0.2129  in  research group, 0.0364  in control group;The  interquartile range of mRNA relative expression of substance P was  0.6954 in  research group,  0.1674  in control group; The percentage of positive staining area of the protein of substance P   was 25.6531±7.0761   in  research group, 20.2915±4.3665   in control group. The mRNA relative expression  and the percentage of positive staining area of substance P were significantly higher in research group than those in control group (P<0.05). Conclusions: The increased expression of substance P in uterosacral ligament of endometriosis patients may be the cause of onic pelvic pain.

语种: 中文
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内容类型: 学位论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/125370
Appears in Collections:北京大学深圳医院_学位论文

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作者单位: 北京大学深圳医院

Recommended Citation:
胡艳. 1 POP阴道壁组织基质金属蛋白酶与胶原的表达及意义 2 P物质在内异症慢性盆腔痛子宫骶韧带中的表达及意义[D]. 北京大学深圳医院. 北京大学. 2016.
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