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RhoA及Rho激酶在子宫腺肌病患者子宫内膜-肌层交界区平滑肌细胞中的表达及其与痛经的关系

Expression of RhoA and Rho kinase in junctional zone of human adenomyosis and its relationship with dysmenorrheal

摘要:

目的 探讨RhoA及Rho激酶(ROCK Ⅰ)在子宫腺肌病患者子宫内膜-肌层交界区平滑肌细胞中的表达及其与痛经程度的关系.方法 选择2012年3至12月在首都医科大学附属北京妇产医院妇科微创中心因子宫腺肌病行子宫全切除术的32例患者为子宫腺肌病组(其中增殖期内膜18例,分泌期内膜14例);同期因宫颈病变或卵巢肿瘤而行子宫切除的患者29例为对照组(其中增殖期内膜12例,分泌期内膜17例).对两组患者子宫内膜-肌层交界区平滑肌细胞进行分离及原代培养.应用实时荧光定量PCR技术和蛋白印迹法,检测两组及子宫腺肌病组不同程度痛经患者子宫内膜-肌层交界区平滑肌细胞中RhoA和ROCKⅠ mRNA及其蛋白的表达水平.结果 (1)子宫腺肌病组子宫内膜-肌层交界区平滑肌细胞中RhoA和ROCK Ⅰ mRNA表达水平无明显周期性变化,增殖期表达水平(分别为1.41 ±0.16、1.05 ±0.15)稍高于分泌期(分别为1.17±0.25、0.98 ±0.10),分别比较,差异均无统计学意义(P>0.05);对照组RhoA和ROCK Ⅰ mRNA表达水平存在周期性变化,增殖期表达水平(分别为0.93 ±0.10、1.00±0.18)高于分泌期(0.48 ±0.03、0.55±0.05),分别比较,差异均有统计学意义(P<0.05);子宫腺肌病组增殖期和分泌期RhoA和ROCK Ⅰ mRNA的表达水平均高于对照组,分别比较,差异均有统计学意义(P<0.01).(2)子宫腺肌病组与对照组RhoA及ROCKⅠmRNA、蛋白表达水平均呈正相关关系(r=0.67,P<O.01;r =0.48,P<0.01).(3)RhoA和ROCKⅠmRNA、蛋白表达水平在子宫腺肌病组不同程度痛经患者中依次表现为:重度痛经(1.66±0.19、1.32±0.11)>中度痛经(1.28 ±0.12、1.09 ±0.08)>轻度痛经(0.93 ±0.09、0.81 ±0.06),两两分别比较,差异均有统计学意义(P<0.05).结论 子宫腺肌病患者子宫内膜-肌层交界区RhoA及ROCKⅠ呈高表达,与患者痛经程度呈正相关,但不随月经周期变化,提示,RhoA及ROCKⅠ持续性高表达可能导致子宫的异常收缩,从而参与子宫腺肌病痛经的发生.

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Objective To investigate the expression of RhoA and Rho kinase in junctional zone (JZ) of adenomyosis and normal myometrium and explore its relationship with severity of dysmenorrheal.Methods From Mar.to Dec.2012,32 cases with adenomyosis undergoing hysterectomy were enrolled as adenomyosis group including 18 cases with proliferative endometrium and 14 cases with secretory endometrium matched with 29 cases with hysterectomy due to cervical disease and ovarian tumor as control group including 12 cases with proliferative endometrium and 17 cases with secretory endometrium in Beijing Obstetrics and Gynecology Hospital Affiliated to Capital Medical University.JZ smooth muscle cells were isolated and cultured immediately after the operation.The expression of mRNA and protein of RhoA and ROCK Ⅰ in JZ in two groups were measured by real-time fluorescence quantitative RT-PCR and western blot.Results (1) The mRNA expression of RhoA and ROCK Ⅰ in JZ of adenomyosis group did not show cyclic change.In proliferative phase,the expression of RhoA and ROCK Ⅰ (1.41 ±0.16,1.05 ±0.15) was not significantly higher than that in secretory phase (1.17 ± 0.25,0.98 ± 0.10) (P > 0.05).While JZ in control group,it showed obviously cyclic change.The expression level of them in proliferative phase(0.93 ±0.10,1.00 ± 0.18) was significantly higher than that in secretory phase (0.48 ± 0.03,0.55 ± 0.05) (P <0.05) ; It also showed that expressions of RhoA and ROCK Ⅰ in adenomysis group were significant higher than those in the control (P < 0.05).(2) The mRNA and protein expression of RhoA and ROCK Ⅰ was positively correlated in each of two groups (r =0.48,P < 0.01 ; r =0.67,P < 0.01).(3) The expression of RhoA and ROCK Ⅰ were 1.66 ±0.19,1.32 ±0.11 in severe dysmenorrheal,1.28 ±0.12,1.09 ±0.08 in moderate dysmenorrheal and 0.93 ± 0.09,0.81 ± 0.06 in mild dysmenorrheal,it all reached statistical difference when compared the other group.(All P < 0.05).Conclusions The expressions of RhoA and ROCK Ⅰ in JZ in adenomyosis group were higher than those in control group,and positively correlated with the severity of dysmenorrheal in adenomysis group,but it does not change with the menstrual cycle.High expression of RhoA and ROCK Ⅰ might be involved in abnormal contraction of uterine myometrium and correlated with the dysmenorrheal in adenomysis.

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