滋肾育胎丸对肾虚—薄型子宫内膜机制的研究

发布时间：2018-03-22 11:57

本文选题：肾虚-薄型子宫内膜　切入点：滋肾育胎丸　出处：《广州中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:1.分析在滋肾育胎丸影响下,肾虚-薄型子宫内膜不孕患者其月经量、子宫内膜厚度、中医症候的变化,探索中医药对薄型子宫内膜的治疗。分析在滋肾育胎丸影响下miR-135a、miR-196b对子宫内膜HOXA10及其下游基因整合素β 3、EMX2基因的调控作用,以期阐明滋肾育胎丸治疗薄型子宫内膜的靶点及药效机制,补充滋肾育胎丸的临床适用范围。方法:1.临床实验:选择90例肾虚型-薄型子宫内膜患者,采用随机数字方法分为滋肾育胎丸组,戊酸雌二醇组,戊酸雌二醇+滋肾育胎丸组30例。戊酸雌二醇组于月经第5日起予戊酸雌二醇2mg,日1次,连续10d;滋肾育胎丸组自月经第5天开始予滋肾育胎丸,口服,淡盐水或蜂蜜水送服。一次5克(约三分之二瓶盖),一日3次。服用至月经来潮停药;戊酸雌二醇+滋肾育胎丸组予戊酸雌二醇+滋肾育胎丸加减,用量用法同前。三组皆以3个月为1疗程,观察三组临床症状积分变化,月经量变化,子宫内膜厚度,妊娠率评估。2.动物实验:将28只雌性SD大鼠随机分为4组,分别为大鼠空白组,肾虚—薄型子宫内膜大鼠空白组,肾虚—薄型子宫内膜大鼠阿斯匹林组,肾虚—薄型子宫内膜大鼠滋肾育胎丸组。予药物干预后,应用qRT-PCR检测技术,检测大鼠子宫内膜miR-135a、miR-196b、HOXA10基因、ITGβ 3基因、EMX2基因的表达变化。结果:1.中医症候积分比较,治疗前三组中医症候积分比较无显著性差异(P=0.6350.05),治疗后有显著性差异(P=0.0000.01),三组间治疗前后积分差值有显著性差异(P=0.0000.001)组内自身治疗前后积分比较均有显著差异(P=0.0000.01)。2.中医症候综合疗效:90例病例中,西药组总有效率为6.67%,中西医组总有效率100%,中医组总有效率70%。3.月经改善情况:三组病例治疗前后月经量积分比较有显著差异(t=-6.407,P=0.0000.01)。4.子宫内膜厚度的改善:三组病例治疗后子宫内膜厚度均较治疗前明显增厚。5.造模组子宫内膜EMX2基因表达较空白模型组、阿司匹林组、滋肾育胎丸组均增高(P0.05),而滋肾育胎丸组子宫内膜EMX2基因表达较其余各组低(P0.05)。6.造模组整合素β 3基因表达较空白对照组、滋肾育胎丸组低(P0.05),滋肾育胎丸组整合素β 3表达较造模组、阿司匹林组表达高(P0.05)。7.滋肾育胎丸组的HOXA10基因相对表达量较空白模型组、阿司匹林组、造模组明显偏高(P0.05),其中造模组的HOXA10基因相对表达量最低(P0.05)。8.与造模组比较,空白对照组,阿司匹林组,滋肾育胎丸组的miR-135a相对表达量均较造模组低,其中,滋肾育胎丸高剂量组的最低(P0.05)。9.与造模组比较,其余各组的miR-196b相对表达量均较造模组低(P0.05)。10.HOXA10基因与EMX2基因表达显著负相关(r=-0.683,P0.01);HOXA10基因与ITGβ 3基因表达显著正相关(r=0.714,P0.01)。11.miR-135a 与 HOXA10 基因负相关性(r=-0.683,P0.01);miR-135a 和 EMX2基因表达高度正相关(r=0.886,P0.01);miR-135a和ITGβ 3基因表达呈现显著负相关(r=-0.722,P0.01)。12.miR-196b 与 HOXA10 基因表达呈显著负相关(r=-0.736,P0.01);miR-196b 与 EMX2基因表达显著相关(r=0.778,P0.01);miR-196b 和 ITGβ 3 基因表达呈现负负相关(r=-0.778,P0.01)。结论:1、本研究临床观察结果显示滋肾育胎丸联合雌激素能有效改善肾虚-薄型子宫不孕患者的子宫内膜厚度,月经量,中医临床症状等,其效果优于单纯使用雌激素治疗。提示滋肾育胎丸通过补肾,对肾虚-薄型子宫不孕患者有治疗作用。2、本研究动物实验结果显示滋肾育胎丸能增强大鼠子宫内膜ITGβ 3、HOXA10基因的表达;其对EMX2、miR-135a、miR-196b基因抑制作用优于阿司匹林组、雌激素组。提示滋肾育胎丸治疗肾虚-薄型子宫内膜大鼠,其可能通过下调miR-135a,miR-196b,上调HOXA10基因的表达,从而调节靶基因EMX2、ITGβ3,以此改善肾虚-薄型子宫内膜大鼠的子宫内膜容受性。
[Abstract]:Objective: to analysis the 1. effects of Zishen Yutai Pill on kidney deficiency, - thin endometrium of infertile patients with menstrual volume, endometrial thickness, change of TCM syndrome, to explore the traditional Chinese medicine therapy on thin endometrium. In the analysis of Zishenyutai pill under the influence of miR-135a, miR-196b on the uterus HOXA10 integrin and its downstream gene beta 3, regulation of EMX2 gene, in order to clarify the target of Zishenyutai pill in the treatment of thin endometrium and pharmacodynamics, clinical application of Zishenyutai pill. Methods: 1. clinical trials: 90 patients with kidney deficiency - thin endometrium patients were randomly into. Kidney Yutai pill group, estradiol valerate group, estradiol valerate + Zishenyutai pill group. 30 cases of estradiol valerate group on the fifth day of menstruation given estradiol valerate 2mg, 1 times a day, continuous 10d; Zishenyutai pill group from the fifth day of menstruation for Zishenyutai pill. Oral, Salt water or honey water delivery service. A 5 grams (about 2/3 bottle), 3 times a day. Take to menstrual withdrawal; estradiol valerate + Zishenyutai pill group were given estradiol valerate + Zishenyutai pill, usage as before. The three groups were both 3 months was 1 course of treatment, observe three groups of clinical symptom changes, menstrual changes, endometrial thickness and pregnancy rate assessment.2. animal experiment: 28 female SD rats were randomly divided into 4 groups, respectively. Rats in the blank group, deficiency of kidney - thin endometrium of rat blank group, deficiency of kidney - thin endometrium of rats in aspirin group kidney, kidney - thin endometrium of rat Yutai pill group. For drug intervention, application of qRT-PCR detection technology, detection of rat endometrial miR-135a, miR-196b, HOXA10 gene, ITG beta 3 gene expression of EMX2 gene. Results: compared with the integral 1. TCM symptoms in three groups before treatment, TCM syndrome integral comparison No significant difference (P=0.6350.05) after treatment, there was significant difference (P=0.0000.01), before and after treatment between the three groups of integral difference have significant difference (P=0.0000.001) significant differences within groups before and after treatment scores were (P=0.0000.01).2. TCM syndrome curative effect in 90 cases, western medicine group the total effective rate was 6.67%. Western medicine group the total efficiency of 100%, the total efficiency of TCM group 70%.3.: menstruation in three groups before and after the treatment of menstruation integral had significant difference (t=-6.407, P=0.0000.01).4. endometrial thickness improved: three groups of patients after treatment of endometrial thickness were significantly thickened.5. model of endometrial EMX2 gene expression compared with the model group, aspirin group, Zishenyutai pill group were increased significantly (P0.05), and Zishenyutai pill group endometrial EMX2 gene expression is low in other groups (P0.05).6. model of integrin beta 3 gene Expression compared with blank control group, Zishenyutai pill group (P0.05), Zishenyutai pill group of integrin beta 3 expression compared with the model group, the high expression of aspirin group (P0.05).7. Yu Zishen relative expression compared with the model group, HOXA10 gene of fetal pill group and aspirin group (model group was significantly higher P0.05), the HOXA10 gene expression of the model group (P0.05.8.) the lowest compared with the model group, blank control group, aspirin group, Zishenyutai pill group, the relative expression level of miR-135a was low compared with that in the model, the Zishenyutai pill high dose group (P0.05) compared with the lowest.9. the model group and the other groups of miR-196b expression were lower than model group (P0.05) and low expression of.10.HOXA10 gene and EMX2 gene significantly negative correlation (r=-0.683, P0.01); the expression of HOXA10 gene and ITG beta 3 gene significantly positive correlation (r=0.714, P0.01).11.miR-135a and HOXA10 gene negative correlation (r=-0.683, P0.01) MiR-135a; the expression of EMX2 gene and high positive correlation (r=0.886, P0.01); miR-135a and ITG beta 3 gene expression showed a significant negative correlation (r=-0.722, P0.01).12.miR-196b and HOXA10 gene expression were negatively correlated (r=-0.736, P0.01); the expression of miR-196b and EMX2 gene significantly correlated (r=0.778, P0.01); miR-196b and ITG beta a related expression showed 3 gene (r=-0.778, P0.01). Conclusion: 1. The study of the clinical observation showed that Zishenyutai pill combined with estrogen can effectively improve kidney - thin infertility uterine endometrial thickness, menstrual volume, clinical symptoms, the effect is better than the simple use of estrogen therapy. Tips for Zishenyutai the therapeutic effect of Bushen pill,.2 of kidney - thin uterine infertility patients, the study of animal experiment results show that Zishenyutai pill can enhance rat endometrial ITG beta 3, HOXA10 gene expression of EMX2, miR-135a; MiR-196b gene, inhibitory effect than aspirin group, estrogen group. Prompt treatment of kidney - nourishing kidney Yu Tai Wan thin endometrium of rats, which may be through the downregulation of miR-135a, miR-196b, up regulation of HOXA10 gene expression, thereby regulating the target gene EMX2, ITG beta 3, in order to improve the deficiency of the kidney - thin endometrium of rat uterine receptivity.

【学位授予单位】：广州中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R271.9

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