本文選題：NF-κB信號通路 + 腎氣虛證��； 參考：《山東中醫(yī)藥大學》2016年碩士論文

【摘要】：目的:研究二至天癸顆粒對腎氣虛型反復種植失敗患者臨床妊娠率及外周血NF-κB信號轉導通路中關鍵因子NF-κB、MCP-1的干預調節(jié)作用,探討腎氣虛致反復種植失敗在NF-κB信號轉導通路水平可能的發(fā)生機制以及補腎中藥在胚胎植入母胎界面免疫耐受中的調節(jié)作用。方法:本研究將符合診斷及辨證標準行體外受精-胚胎移植(IVF-ET)治療的腎氣虛反復種植失敗患者70例,按照門診就診時間先后順序編號,奇數(shù)號分入治療組,偶數(shù)號分入安慰劑組,另選取因男方因素行IVF的健康婦女35例(非腎氣虛組)。治療組在服用二至天癸顆粒3個周期后聯(lián)合常規(guī)IVF治療,安慰劑組及非腎氣虛組服用安慰劑3個周期并聯(lián)合常規(guī)IVF治療,觀察腎氣虛反復種植失敗患者的腎虛證候積分變化;受精率;優(yōu)質胚胎率;生化妊娠率;臨床妊娠率。采用酶聯(lián)免疫吸附方法(ELISA)檢測三組抗絨毛膜促性腺激素抗體(Ah CGAb)、抗心磷脂抗體(ACA-Ig G)、抗精子抗體(As Ab)、抗子宮內膜抗體(AEMAb)、抗卵巢抗體(AOVAb)水平,采用實時熒光定量PCR(RT-PCR)技術、蛋白質印記(Western-blot)法分別檢測移植日外周血中NF-κB、MCP-1m RNA及NF-κB、MCP-1蛋白的含量;采用ELISA法檢測三組患者移植后7天血清MCP-1濃度。結果:1.二至天癸顆粒治療組患者經治療后腎氣虛癥狀明顯改善,證候積分差異顯著(P0.01);兩組治療前后腎氣虛證候積分差值有顯著統(tǒng)計學差異(P0.01)。2.與安慰劑組相比,治療組及非腎氣虛組受精率、優(yōu)質胚胎率、生化妊娠率、臨床妊娠率均較高,有統(tǒng)計學差異(P0.05);治療組與非腎氣虛組之間無統(tǒng)計學差異(P0.05)。3.與非腎氣虛組相比,安慰劑組及治療組Ah CGAb、ACA-Ig G、As Ab、AEMAb、AOVAb陽性率均較高,有統(tǒng)計學差異(P0.05);安慰劑組與治療組之間無統(tǒng)計學差異(P0.05)。4.分析三組患者外周血NF-κB信號轉導通路中關鍵效應分子NF-κB及其下游因子MCP-1的表達,發(fā)現(xiàn):與安慰劑組相比,非腎氣虛組及治療組移植日NF-κB、MCP-1蛋白含量均下降,NF-κBm RNA、MCP-1m RNA表達均下調,移植后7天血清MCP-1含量降低,有統(tǒng)計學差異(P0.05);治療組與非腎氣虛組相比,NF-κB、MCP-1蛋白含量、NF-κBm RNA、MCP-1m RNA表達水平、血清MCP-1含量無統(tǒng)計學差異(P0.05)。5.進一步將患者分為妊娠組與未妊娠組,兩組相比,發(fā)現(xiàn):妊娠組Ah CGAb、ACA-Ig G、As Ab、AEMAb、AOVAb陽性率及血清MCP-1濃度均低于未妊娠組,有顯著統(tǒng)計學差異(P0.01)。結論:1.二至天癸顆�？擅黠@改善腎氣虛型反復種植失敗患者的腎虛狀態(tài),提高其受精率及優(yōu)質胚胎率,改善臨床妊娠率。2.生殖抗體可能是影響妊娠環(huán)境的重要免疫因素。3.NF-κB、MCP-1m RNA及蛋白的高表達可能與反復種植失敗的發(fā)病機制有關,血清MCP-1對于預測早期胚胎移植狀況可能具有重要的臨床意義。4.二至天癸顆粒可能是通過抑制NF-κB信號轉導通路,下調MCP-1因子表達,降低母胎界面免疫排斥作用,從而提高種植率,改善腎氣虛反復種植失敗患者的妊娠結局。
[Abstract]:Objective: to study the effect of Erzhi Tiangui granule on the clinical pregnancy rate and the intervention and regulation of the key factor NF- 魏 B MCP-1 in peripheral blood NF- 魏 B signal transduction pathway in patients with repeated implantation failure of kidney-qi deficiency type. To explore the possible mechanism of repeated implantation failure induced by deficiency of kidney qi at the level of NF- 魏 B signal transduction pathway and the regulatory role of traditional Chinese medicine for tonifying the kidney in the immune tolerance of embryo implantation interface. Methods: in this study, 70 patients with kidney qi deficiency were divided into treatment group according to the order of outpatient visit time and odd number, which met the criteria of diagnosis and syndrome differentiation and were treated with IVF-ETS in vitro fertilization and embryo transfer. Even numbers were divided into placebo group and 35 healthy women who were treated with IVF because of male factors (non-kidney qi deficiency group). The patients in the treatment group were treated with routine IVF after taking Erzhi Tiangui granule for 3 cycles, the placebo group and non-kidney-qi deficiency group were treated with placebo for 3 cycles and combined with routine IVF, and the changes of syndrome score of kidney deficiency in the patients with failed implantation of kidney qi deficiency were observed. Fertilization rate; High quality embryo rate; biochemical pregnancy rate; Clinical pregnancy rate. Enzyme linked immunosorbent assay (Elisa) was used to detect the levels of anti-chorionic gonadotropin antibody (ACCH), anti-chorionic gonadotropin antibody (AHG), anti-cardiolipid antibody (ACA -Ig GG), anti-sperm antibody (as Abn), anti-endometrium antibody (AEMAbn), anti-ovarian antibody (AOVAb), and real-time fluorescence quantitative PCRRT-PCR (RT-PCR) technique. Protein imprinting Western-blot method was used to detect the contents of NF- 魏 BmP-1m RNA and NF- 魏 BmP-MCP-1 in peripheral blood of the patients on the day of transplantation, and the serum MCP-1 concentration of the three groups was detected by ELISA method on the 7th day after transplantation. The result is 1: 1. After treatment, the symptoms of deficiency of kidney qi were obviously improved in Erzhi Tiangui granule group, and the difference of syndrome score was significant (P 0.01). Compared with placebo group, the fertilization rate, high quality embryo rate, biochemical pregnancy rate and clinical pregnancy rate were higher in treatment group and non-kidney-qi deficiency group, and there was no statistical difference between treatment group and non-kidney-qi deficiency group. Compared with non-kidney-qi deficiency group, the positive rate of Ah CGAbACA-Ig GG as AbAEMAbAb-AOVAb in placebo group and treatment group was higher than that in non-kidney-qi deficiency group (P 0.05), but there was no significant difference between placebo group and treatment group (P 0.05. 4). The expression of NF- 魏 B and its downstream factor MCP-1 in peripheral blood NF- 魏 B signal transduction pathway was analyzed in three groups. It was found that the expression of NF- 魏 B mRNA MCP-1 protein in non-kidney-qi deficiency group and treatment group was decreased and the expression of NF- 魏 Bm RNA-MCP-1m RNA was down-regulated on the day of transplantation. After 7 days of transplantation, the content of serum MCP-1 decreased with statistical difference (P0.05), and the protein content of NF- 魏 BmRNA-MCP-1 in treatment group was higher than that in non-kidney-qi deficiency group (P < 0.05), and the expression level of NF- 魏 Bm RNA-MCP-1 RNA was not significantly different in treatment group compared with that in non-kidney-qi deficiency group (P < 0.05). The patients were further divided into gestational group and non-pregnant group. It was found that the positive rate of Ah CGAbACA-Ig AEM AbAbAAOVAb and serum MCP-1 concentration in pregnancy group were significantly lower than those in non-pregnant group (P 0.01). Conclusion 1. Erzhi Tiangui granule can obviously improve the kidney deficiency of the patients with kidney qi deficiency, improve the fertilization rate and the high quality embryo rate, and improve the clinical pregnancy rate. 2. The high expression of MCP-1m RNA and protein may be related to the pathogenesis of repeated implantation failure, and serum MCP-1 may have important clinical significance in predicting the status of early embryo transfer. Erzhitiangui granule may be by inhibiting NF- 魏 B signal transduction pathway, down-regulating the expression of MCP-1 factor, reducing the maternal and fetal interface immune rejection, so as to improve the implantation rate and improve the pregnancy outcome of patients with repeated failed implantation of kidney qi deficiency.
【學位授予單位】：山東中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R271.9

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