卵巢过度刺激综合征诊断标志物的筛选及“瘀滞”论治疗的临床效应评估

发布时间：2018-05-26 19:21

本文选题：卵巢过度刺激综合征 + 解瘀滞中药　；参考：《南京中医药大学》2016年博士论文

【摘要】：目的：1.临床观察从“瘀滞”论入手治疗OHSS患者的有效性；2.探讨解“瘀滞”中药对OHSS大鼠模型血管通透障碍的作用及机制；3.筛选有较高特异性和灵敏度的OHSS血清诊断标志物。方法：1.选择68个符合标准的受试者作为研究对象,分为对照组、中药组、西药组,COS治疗后,未患OHSS的受试者设为对照组,发生OHSS的受试者随机分为中药组、西药组,分别给予中药和西药治疗。中药组给予解瘀滞中药治疗,西药组则给予阿司匹林治疗,治疗时间一周。检测周期妊娠结局、排卵率、周期放弃率、卵泡增大的数量以及各组受试者血清中E2、VEGF、IL-6水平。2.按照Ujioka报道的方法构建OHSS大鼠病理模型。将66只大鼠随机分为正常对照组、OHSS组、西药组、中药组低、中、高剂量组。按照人与动物体表面积换算给药剂量给予大鼠药物,检测大鼠血清、腹水NO、VEGF、IL-6等的水平,对大鼠体重和卵巢重量进行检测,比较各组间的差异。3.对OHSS患者和非OHSS患者的血清进行质谱分析差异蛋白,对差异大且有涉及OHSS生物过程的蛋白进一步通过蛋白免疫印迹和酶联免疫吸附分析,筛选特异性强、准确度高的血清诊断标志物。结果：1.中药组在周期放弃率和增大卵泡数都显著低于西药组。2.血清E2水平、血管内皮生长因子(VEGF)、腹围、盆腔积液等临床指标,中药组都显著低于西药组。3. OHSS组大鼠血清E2、P、VEGF、IL-6水平显著升高,腹腔和卵巢EB水平较其余各组都显著升高。4.中药中、高剂量组大鼠体重、卵巢重量、腹水量、EB、NO水平、VEGF、IL-6水平都显著低于西药组。5.质谱结果筛选到57个表达差异蛋白,与对照组相比,OHSS组有29种蛋白表达上调,而28种蛋白表达明显下调。6.蛋白免疫印迹和酶联免疫吸附试验验证得到三个高差异表达蛋白：结合珠蛋白、纤维蛋白原和脂蛋白脂酶,ROC曲线分析中,三种蛋白对OHSS预测都有较高敏感性和特异性。结论：1.从“瘀滞”论入手治疗OHSS患者,能显著降低患者的多项炎症因子水平,并且提高排卵率、降低周期放弃率,而不影响临床妊娠率,取得较好的疗效。2.动物实验证实解瘀滞中药能够明显降低OHSS大鼠血清中相关炎症因子水平,且对大鼠卵巢重量、腹水有明显的改善作用。3.由质谱筛选和进一步验证得到结合珠蛋白、纤维蛋白原、脂蛋白脂肪酶可以作为血清诊断预测OHSS发生的生物标志物,且具有较高特异性和准确性。
[Abstract]:Purpose 1. Clinical observation on the effectiveness of treating OHSS patients from the view of "stagnation". To investigate the effect and mechanism of traditional Chinese medicine (TCM) on vascular permeability in OHSS rats. Screening OHSS serum diagnostic markers with high specificity and sensitivity. Method 1: 1. Sixty-eight subjects who met the criteria were divided into three groups: control group, traditional Chinese medicine group and western medicine group. The subjects who did not suffer from OHSS were randomly divided into Chinese medicine group and western medicine group. Chinese medicine and western medicine were given respectively. The traditional Chinese medicine group was treated with traditional Chinese medicine and the western medicine group with aspirin for one week. The pregnancy outcome, ovulation rate, cycle abandonment rate, the number of follicular enlargement and the level of E2VEGFN IL-6 in the serum of each group were measured. The pathological model of OHSS rats was established according to the method reported by Ujioka. 66 rats were randomly divided into three groups: normal control group, western medicine group, low, middle and high dosage groups. According to the conversion of human and animal body surface area, the drug was given to rats. The levels of serum and ascites VEGF IL-6 were detected. The body weight and ovarian weight of rats were measured, and the difference between each group was compared. The differential proteins in serum of OHSS patients and non-OHSS patients were analyzed by mass spectrometry, and the proteins involved in the biological process of OHSS were further screened by Western blot and enzyme-linked immunosorbent assay (Elisa). Serum diagnostic markers with high accuracy. The result is 1: 1. The rate of cycle abandonment and the number of follicles increased in the Chinese medicine group were significantly lower than those in the western medicine group. Serum E2 level, vascular endothelial growth factor (VEGF) VEGF, abdominal circumference, pelvic effusion and other clinical indexes were significantly lower in the Chinese medicine group than in the western medicine group. In OHSS group, the serum level of E _ 2P _ (2) P _ (VEGF) IL-6 was significantly increased, and the EB level in abdominal cavity and ovary was significantly higher than that in other groups. In Chinese medicine, the levels of body weight, ovarian weight and ascites volume of EBNO in high dose group were significantly lower than those in western medicine group (P < 0.05). 57 differentially expressed proteins were screened by mass spectrometry. 29 proteins were up-regulated in OHSS group and 28 proteins were down-regulated in OHSS group. Western blot and enzyme-linked immunosorbent assay (Elisa) showed that three highly differentially expressed proteins were highly sensitive and specific to OHSS prediction: globin, fibrinogen and lipoprotein lipase. Conclusion 1. Starting with the theory of "stagnation", the treatment of OHSS patients can significantly reduce the level of multiple inflammatory factors, and increase ovulation rate, reduce cycle abandonment rate, without affecting the clinical pregnancy rate, and obtain a better curative effect. 2. Animal experiments have proved that traditional Chinese medicine can significantly reduce the level of serum inflammatory factors in OHSS rats, and can significantly improve the weight of ovary and ascites in rats. 3. The binding globin, fibrinogen and lipoprotein lipase can be used as biomarkers for the diagnosis and prediction of OHSS by mass spectrometry.
【学位授予单位】：南京中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R271.9

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