Objective To establish rat intrauterine adhesion(IUA)model and check theexpressions of HSP47 protein and HSP47 mRNA in endometrium in rats so as to explore expression of HSP47 in intrauterine adhesion model and influence of adhesion on fertility.Methods Ninety healthy and sexually mature S-D rats were raised,among which 30 were male and 60 were female.Female rats were divided into IUA group(30 cases),sham operation group(15 cases)and normal control group(15 cases)randomly. Intrauterine adhesion models were prepared with phenol glue method.Fifteen rats in IUA group,7 rats in sham operation group and 7 rats in normal control group were killed at 10d after model making operation.Changes of uterine cavity were observed with naked eyes.Pathological changes were investigated with HE staining.Expression of HSP47 in rat endometrium was checked with immunohistochemical staining,and the expression of HSP47 mRNA was detected with real-time PCR.For other female rats,one rat was raised in pairs with one male rat in same cage until they got pregnant naturally.Laparotomy was implemented on pregnant rats on the 10th day of pregnancy,and number of embryos was counted as fertility evaluation index.Results HSP47 was mostly located at cell nucleus.Comparison of expressions of HSP47 and HSP47 mRNA at left side of uterus found that expression levels in IUA group were remarkably higher than those in normal control group and sham operation group(t=9.07,P<0.05).At model side,positive rate of high expression of HSP47 in endometrium was 80% in IUA group,and that was 14.29% in both normal control group and sham operation group.No significant difference existed between sham operation group and normal control group (χ2= 3.19,P > 0.05).HSP47 high expression rate was 14.29% in both normal control group and sham operation group. Expressions of HSP47 and HSP47 mRNA at right side of uterus in IUA group were obviously higher than those in normal group and sham operation group(t=10.14,P<0.05).HSP47 high expression rate at control side in IUA group was 66.67%,and that in both normal group and sham operation group was 14.29%.No significant difference existed between normal group and sham operation group(χ2=0.00,P>0.05).HSP47 high expression rate was 14.29% in both normal group and sham operation group, while expressions of HSP47 and HSP47 mRNA at both sides of uterus were increased in adhesion group,and difference was not statistically significant(t=0.35,P>0.05).Relative expression quantity of HSP47 in endometrium was 6.29 ± 1.09 at left side and 6.59 ± 1.3 at right side.Number of embryos in left side uterus was 0.20 ± 0.40 in IUA group,6.30 ± 1.80 in normal group,and 6.30 ± 2.90 in sham operation group with statistical significance(F=8.83,P<0.001).Embryo number at right side uterus was 4.90 ± 2.60 in IUA group,5.40 ± 1.60 in normal group and 5.90 ± 3.00 in sham operation group,and such difference was not statistically significant(F=0.39,P>0.05).In IUA group,number of embryos at left side uterus was 0.20 ± 0.40 and 4.90 ± 2.60 at right side uterus,and difference was statistically significant(t=6.92,P<0.001).Embryo number at left side uterus in sham operation group was 6.30 ± 2.90 and 6.30 ± 1.80 in normal control group,and embryo number at right side uterus was 5.90 ± 3.00 in sham operation group and 5.40 ± 1.60 in normal control group,and difference in embryo number between sham operation group and normal group was not statistically significant(t= 0.57,P> 0.05).Conclusion Expression of HSP47 has been remarkably increased in rats with intrauterine adhesion and fertility of rats with intrauterine adhesion is significantly weakened.%目的 建立大鼠宫腔粘连模型,检测各组大鼠子宫内膜组织中 HSP47蛋白和 HSP47 mRNA 的表达量,探讨HSP47在宫腔粘连模型中的表达及宫腔粘连模型对生育力的影响.方法 饲养健康性成熟S-D大鼠90只,雄性大鼠30只,雌性大鼠60只,将雌性大鼠随机分为造模组(30只)、假手术组(15只)、正常组(15只),苯酚胶浆法建立宫腔粘连模型.造模术后10d,取造模组15只、假手术组7只、正常7只处死,肉眼观察其宫腔改变,HE染色观察病理变化,免疫组化染色检测大鼠子宫内膜HSP47的表达,Real-time PCR检测HSP47 mRNA的表达.剩余大鼠一对一与雄性大鼠合笼饲养,使其自然妊娠,妊娠第10天开腹计数子宫内胚胎数目作为评价生育力的指标.结果 HSP47主要定位于细胞核,左侧子宫 HSP47和 HSP47 mRNA的表达比较发现,造模组明显高于正常组和假手术组(t=9.07,P<0.05).在造模侧,HSP47在造模组子宫内膜的高表达阳性率为80%,而在正常组和假手术组的高表达阳性率均为14.29%.正常组和假手术组相比,差异无统计学意义(χ2= 3.19,P>0.05),HSP47在正常组和假手术组高表达阳性率均为14.29%.右侧子宫HSP47和HSP47mRNA的表达,造模组也明显高于正常组和假手术组(t=10.14,P<0.05).在对照侧,HSP47在子宫内膜在造模组的高表达阳性率为66.67%,而正常组和假手术组高表达阳性率均为14.29%.正常组和假手术组相比差异无统计学意义(χ2=0.00,P>0.05),HSP47在正常组和假手术组子宫内膜高表达阳性率均为14.29%.宫腔粘连组左、右侧子宫 HSP47和 HSP47mRNA的表达均增高,差异没有统计学意义(t=0.35,P>0.05).HSP47在该组子宫内膜的相对表达量左侧为(6.29 ± 1.09),右侧为(6.59 ± 1.3).妊娠胚胎数,左侧子宫造模组为(0.20 ± 0.40)个,正常组为(6.30 ± 1.80)个,假手术组为(6.30 ± 2.90)个,差异有统计学意义(F=8.83,P<0.001).右侧子宫造模组为(4.90 ± 2.60)个,正常组为(5.40 ± 1.60)个,假手术组为(5.90 ± 3.00)个,差异无统计学意义(F=0.39,P>0.05).宫腔粘连组左侧子宫为(0.20 ± 0.40)个,右侧子宫为(4.90 ± 2.60)个,差异有统计学意义(t=6.92,P<0.001).左侧子宫妊娠胚胎数假手术组为(6.30 ± 2.90)个,正常组为(6.30 ± 1.80)个,右侧子宫假手术组为(5.90 ± 3.00)个,正常组为(5.40 ± 1.60)个,假手术组和正常组差异无统计学意义(t=0.57,P>0.05).结论 HSP47在宫腔粘连大鼠子宫内膜组织中表达明显增高,宫腔粘连的大鼠生育力明显降低.
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