17α-羟基孕酮己酸酯和醋酸甲羟孕酮预防孕鼠感染性早产的机制

Effect of 17α-hydroxyprogesterone caproate and medroxyprogesterone acetate on inflammation-induced preterm birth of mouse model

目的通过研究17α-羟基孕酮己酸酯（17-αydroxyprogester。necaproate，17P）和醋酸甲羟孕酮（medroxyprogesteroneacetate，MPA）对早产小鼠子宫肌层及胎盘组织环氧合酶-2（cycl00xygenase-2，COX-2）和肿瘤坏死因子-α（tumornecrosisfactor-α，TNF-α）表达的影响，探讨孕激素预防感染性早产的作用机制。方法选择清洁级雌性CD-1小鼠30只，于妊娠第15天，随机分为对照组、内毒素脂多糖（1ipop01ysaccharides，LPS）组、17Plmg＋LPS组、17P2mg＋LPS组、MPA1mg＋LPS组、二甲基亚砜（dimethylsulfoxide，DMSO）＋LPS组，每组5只。治疗组均于皮下注射相应剂量的孕激素1h后腹腔注射LPS，6h后取小鼠子宫肌层及胎盘组织，用实时荧光定量聚合酶链反应技术和免疫组织化学方法检测子宫肌层及胎盘组织COX-2和TNF-α mRNA及蛋白的表达。采用ANOVA进行统计学分析，两两比较用LSD法。结果1．各组小鼠子宫肌层及胎盘组织中COX-2mRNA和TNF-α mRNA的相对表达量的比较：（1）各给药组孕鼠子宫肌层及胎盘组织COX-2mRNA和TNFlamRNA表达均显著高于对照组（P〈0．05）。（2）17P1mg＋LPS组、17P2mg＋LPS组和MPA1mg＋LPS组小鼠子宫肌层COX-2mRNA表达分别为11．410±3．931、8．352±3．209和11．920±2．905，均显著低于LPS组（20．540±4．147）和DMSO组＋LPS组（18．620±4．156）（P〈O．05），而TNF-αmRNA也低于LPS组和DMSO组＋LPS组，但差异无统计学意义（P〉O．05）。（3）17P1mg＋LPS组、17P2mg＋LPS组、MPA1mg＋LPS组胎盘组织COX-2 mRNA表达分别为10．864±3．777、7．084±L667和11．830±3。652，均显著低于LPS组（18．920±4．106）和DMSO组＋LPS组（23．820±7．554）（P〈0．05）。（4）17Plmg＋LPS组、17P2mg＋LPS组、MPA1mg＋LPS组胎盘组织TNF-αITIRNA表达分别为14．340±1．618、11．488±2．910和13．040±2．982，均显著低于LPS组（24．240±7．059）和DMs0组＋LPS组（23．040±5．896）（P〈0．05）。2．各组小鼠胎盘组织COX-2和TNF-α蛋白表达的比较：（1）各给药组胎盘组织中COX-2蛋白的表达和TNF-α蛋白的表达明显高于对照组（P〈0．05）。（2）17P1mg＋LPS组、17P2mg＋LPS组和MPA1mg＋LPS组cox-2蛋白的表达分别为14360．92±1766．01、13340．18±965．35、12870．81±1521．97，明显低于LPS组（16426．64±1823．87）和DMS0组＋LPS组（16761．23±2388．17）（P〈0．05）；但3个治疗组之间差异无统计学意义（P〉0．05）。（3）17P1mg＋LPS组、17P2mg＋LPS组、MPA1mg＋LPS组TNF-α蛋白的表达分别为22750．96±4656．68、22766．24±3500．34和20770．01±3318．48，显著低于LPS组（26204．49±5090．34）和DMS0组＋LPS组（27346．18±3269．30）（P〈0．05），而3组之间差异无统计学意义（P〉0．05）。产小鼠子宫和胎盘组织COX-2及胎盘中TNF-α的表达有抑制作用结论17P和MPA对感染性早，可能是其预防早产的机制之一。

Objective To study the effect of 17α-hydroxyprogesterone caproate （17P） andmedroxyprogesterone acetate （MPA） on expression of tumor necrosis factor-α （TNF-α）and cyclooxygenase-2 （COX-2） in placenta and uterine myometrium of inflammation-induced preterm birth mouse model to investigate the mechanism o{ preventing inflammation-induced preterm birth by progestogen. Methods Thirty clean CD-1 mice were divided into 6 groups （5 mice in each group） at 15th day of gestation： control group, lipopolysaccharides （LPS） group, 17P 1 mg-αLPS group, 17P 2 rag＋ LPS group, MPA 1 mg＋LPS group and dimethyl sulfoxide （DMSO） ＋LPS group. Progestogens at different dosage were administered 1 h before LPS and 6 h after LPS administration. After these mice were sacrificed, TNF-α and COX-2 levels in the myometrium and placenta were detected by real-time PCR and immunohistoehemistry. Data were analyzed by ANOVA, and comparisons between groups were adopted LSD method. Results 1. The comparison of relative expression of COX-2 mRNA and TNF-α mRNA in myometrium and placenta among groups： （1） Expressions of COX-2 mRNA and TNF-α mRNA in myometrium and placenta in the study groups were obviously higher than those of control group （P〈0.05）. （2） COX-2 mRNA expression in myometrium of 17P 1 mg＋LPS group （11. 410±3. 931）, 17P 2 mg＋LPS group （8. 352±3. 209） and MPA 1 mg-kLPS group （11. 920 ± 2. 905） were obviously lower than that of LPS group （20. 540 ± 4. 147） and DMSO^LPS group （18. 620±4. 156）（P〈0.05, respectively）; although TNF-α mRNA expression had similar trends among these groups, there were no statistical significance （P〈0.05, respectively）. （3） COX-2 mRNA expression in placenta of 17P 1 mg＋LPS group （10. 864±3. 777）, 17P 2 mg＋LPS group （7. 084±1. 667） and MPA 1 mg＋LPS group （11. 830±3. 652） were obviously lower than that of LPS group （18. 920±4. 106） and DMSOTLPS group （23. 820±7. 554） （P〈0. 05, respectively）. （4） TNF-α mRNA expression in placenta in 17P 1 mg＋LPS group （14. 340±1. 618）, 17P 2 mg-1 LPS group （11. 488 ± 2. 910） and MPA 1 mg＋ LPS group （13. 040 ± 2. 982） were obviously lower than that of LPS group （24. 240±7. 059） and DMSO＋LPS group （23. 040~5. 896） （P〈0. 05, respectively）. 2. The comparison of protein expression of COX-2 and TNF-α in placenta among groups： （1） The expression of COX-2 protein in placenta of the study groups was significantly higher than that of control group （P〈0.05）. （2） There were no differences among the COX-2 protein expression of 17P 1 mg＋LPS group （14 360.92 ±1766.01）, 17P 2 mg＋LPS group （13 340.18 ± 965.35） and MPA 1 mg＋LPS group （12 870.81 ± 1521.97） （P〉0.05）, while the COX-2 protein expressions of them were significantly lower than that of LPS group （16 426.64±1823.87） and DMSO＋LPS group （16 761. 23± 2388.17）（P〈0.05, respectively）. （3） There were no differences among the TNF-α protein expression of 17P 1 mg-[LPS group （22 750. 96±4656.68）, 17P 2 mg＋ LPS group （22 766. 24±3500.34） and MPA 1 mg＋LPS group （20 770. 01±3318.48）（P〉0.05）, while the TNF-α protein expressions of them were significantly lower than that of LPS group （26 204.49±5090.34） and DMSO＋LPS group （27 346.18±3269.30）（P〈0.05, respectively）. Conclusions 17P and MPA might prevent the preterm parturition of inflammation-induced mouse model by inhibiting inflammation cytokines and prostaglandins.