IFI16在子癇前期胎盤組織和血清中的表達(dá)及與子癇前期發(fā)病的相關(guān)性
發(fā)布時(shí)間：2018-09-01 07:21
【摘要】：子癇前期是妊娠期特有的疾病之一,以妊娠20周以后新發(fā)的持續(xù)性血壓升高并伴有明顯蛋白尿?yàn)橹饕卣?在全世界的發(fā)病率約為3%~8%,嚴(yán)重影響母嬰健康。然而到目前為止,子癇前期的確切發(fā)病機(jī)制仍不清楚。現(xiàn)在研究認(rèn)為,內(nèi)皮細(xì)胞損傷是子癇前期發(fā)病的中心環(huán)節(jié)。人γ干擾素誘導(dǎo)蛋白16(Interferon-inducible protein 16,IFI16)屬于HIN-200家族成員,主要存在于造血細(xì)胞、成纖維細(xì)胞、上皮細(xì)胞、內(nèi)皮細(xì)胞、滋養(yǎng)細(xì)胞等細(xì)胞內(nèi)。最近研究發(fā)現(xiàn),IFI16蛋白也可存在于自身免疫性疾病患者血清中,且血清中高水平的IFI16可能與內(nèi)皮細(xì)胞損傷有關(guān)。氧化應(yīng)激、各種炎性因子和細(xì)胞密度可觸發(fā)IFI16表達(dá)上調(diào)�？紤]到子癇前期胎盤組織處于強(qiáng)氧化應(yīng)激和高炎性反應(yīng)狀態(tài),內(nèi)皮細(xì)胞損傷是其發(fā)病的中心環(huán)節(jié),推測(cè)IFI16蛋白可能與子癇前期發(fā)病有關(guān)。然而,目前關(guān)于IFI16蛋白在子癇前期中的研究尚比較少見。目的本研究通過檢測(cè)IFI16在正常組和子癇前期組胎盤組織和血清中的表達(dá),并對(duì)血清IFI16水平與內(nèi)皮細(xì)胞損傷相關(guān)的檢測(cè)指標(biāo),如血壓、24h尿蛋白、血清總蛋白、白蛋白、尿素、尿酸、肌酐、胱抑素C、β2-微球蛋白、α1-微球蛋白、乳酸脫氫酶和內(nèi)皮素-1(Endothelin-1,ET-1)等指標(biāo)之間的相關(guān)性進(jìn)行分析,最后利用受試者工作曲線(receiver operator characteristic curve,ROC曲線)及約登指數(shù)評(píng)判血清IFI16水平在預(yù)測(cè)子癇前期中的價(jià)值,從而探討IFI16與子癇前期發(fā)病的相關(guān)性,并為該指標(biāo)的臨床應(yīng)用提供實(shí)驗(yàn)室依據(jù)。方法收集正常組和子癇前期組孕婦胎盤組織和血清,分別采用免疫組化法、實(shí)時(shí)熒光定量PCR技術(shù)和蛋白印跡法檢測(cè)胎盤組織中IFI16的表達(dá);采用全自動(dòng)生化分析儀檢測(cè)兩組孕婦血清總蛋白、白蛋白、尿素、尿酸、肌酐、胱抑素C、β2-微球蛋白、α1-微球蛋白、乳酸脫氫酶及24h尿蛋白等生化指標(biāo);采用ELISA方法檢測(cè)血清中IFI16和ET-1的水平,并對(duì)血清IFI16水平與各檢測(cè)指標(biāo)之間的相關(guān)性進(jìn)行分析。應(yīng)用SPSS 21.0進(jìn)行數(shù)據(jù)的分析和處理,結(jié)果以均數(shù)±標(biāo)準(zhǔn)差(?x±s)表示,兩組間差異性比較采用獨(dú)立樣本t檢驗(yàn)或卡方檢驗(yàn)(χ2);相關(guān)性分析采用Pearson相關(guān)性檢驗(yàn);ROC曲線用于評(píng)判血清IFI16水平能否作為一種新的生物學(xué)指標(biāo)用于預(yù)測(cè)子癇前期的發(fā)生,并確定相應(yīng)的預(yù)測(cè)界值。以α=0.05為檢驗(yàn)水準(zhǔn)。結(jié)果1兩組孕婦一般臨床資料和檢測(cè)指標(biāo)的比較1.1兩組孕婦一般臨床資料的比較子癇前期組和對(duì)照組孕婦年齡分別為(30.9±4.6)歲和(29.2±4.2)歲,采血孕周分別為(34.2±2.3)周和(34.3±1.0)周,分娩孕周分別為(35.3±2.6)周和(38.6±1.0)周,收縮壓分別為(156.6±9.6)mm Hg和(115.1±9.5)mm Hg,舒張壓分別為(103.9±7.6)mm Hg和(73.6±5.9)mm Hg,新生兒體重分別為(2170.3±309.2)g和(3316.1±456.1)g,24小時(shí)尿蛋白量分別為(2003.5±110.9)mg和(83.2±17.2)mg。經(jīng)統(tǒng)計(jì)學(xué)分析,兩組孕婦年齡和采血孕周相比,差異無統(tǒng)計(jì)學(xué)意義(P0.05);子癇前期組收縮壓、舒張壓以及24小時(shí)尿蛋白定量均高于對(duì)照組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05);子癇前期組分娩孕周和胎兒體重均低于對(duì)照組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。1.2兩組孕婦臨床檢測(cè)指標(biāo)的比較子癇前期組和對(duì)照組孕婦血清總蛋白濃度分別為(56.3±6.8)g/L和(61.6±5.5)g/L,白蛋白濃度分別為(29.8±4.2)g/L和(34.7±2.9)g/L,尿素氮濃度分別為(4.0±0.8)mmol/L和(3.5±0.7)mmol/L,尿酸濃度分別為(317.4±94.5)μmol/L和(263.0±66.8)μmol/L,肌酐濃度分別為(53.6±11.5)μmol/L和(44.6±6.6)μmol/L,β2-微球蛋白濃度分別為(2.3±0.6)mg/L和(1.8±0.4)mg/L,α1-微球蛋白濃度分別為(27.5±7.7)mg/L和(24.7±4.1)mg/L,胱抑素C濃度分別為(1.6±0.4)μmol/L和(1.1±0.2)μmol/L,乳酸脫氫酶濃度分別為(256.6±61.8)U/L和(198.5±30.1)U/L。經(jīng)統(tǒng)計(jì)學(xué)分析,子癇前期組孕婦血清尿素氮、尿酸、肌酐、β2-微球蛋白、α1-微球蛋白、胱抑素C、乳酸脫氫酶水平均高于對(duì)照組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05);子癇前期組孕婦血清總蛋白和白蛋白水平均低于對(duì)照組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。2兩組孕婦胎盤組織中IFI16蛋白的表達(dá)和定位兩組孕婦胎盤滋養(yǎng)細(xì)胞和血管內(nèi)皮細(xì)胞的細(xì)胞質(zhì)中均有IFI16蛋白的表達(dá)。與對(duì)照組相比,IFI16蛋白在子癇前期組胎盤組織中的陽性表達(dá)率明顯增高(IFI16在正常組和子癇前期組胎盤組織中的陽性表達(dá)率分別為43%和78%),兩組比較,差異具有統(tǒng)計(jì)學(xué)意義(χ2=9.671,P0.05)。3兩組孕婦胎盤組織中IFI16 m RNA和蛋白質(zhì)表達(dá)水平的比較子癇前期組胎盤組織中IFI16 m RNA的相對(duì)表達(dá)量(1.65±0.08)高于對(duì)照(0.72±0.05),兩組比較,差異具有統(tǒng)計(jì)學(xué)意義(P0.01);子癇前期組胎盤組織中IFI16蛋白的相對(duì)表達(dá)水平(1.58±0.06)亦高于對(duì)照組(0.56±0.03),差異均具有統(tǒng)計(jì)學(xué)意義(P0.01)。4兩組孕婦血清中IFI16及ET-1水平的比較子癇前期組血清IFI16的濃度為(20.68±8.9)ng/m L,對(duì)照組為(10.08±4.1)ng/m L,兩組比較,差異有統(tǒng)計(jì)學(xué)意義(P0.01);子癇前期組血清ET-1的水平為(41.6±9.3)ng/L,對(duì)照組為(11.0±4.9)ng/L,兩組比較,差異有統(tǒng)計(jì)學(xué)意義(P0.01)。5子癇前期組孕婦血清IFI16水平與各檢測(cè)指標(biāo)之間的相關(guān)性Pearson相關(guān)性分析表明,子癇前期組孕婦血清IFI16水平與孕婦收縮壓(r=0.639,P0.01)、舒張壓(r=0.514,P0.01)、24h尿蛋白定量(r=0.707,P0.01)、胱抑素C(r=0.347,P0.05)和血清ET-1水平(r=0.616,P0.01)呈正相關(guān),與血清白蛋白水平呈負(fù)相關(guān)(r=-0.366,P0.05);與孕婦年齡、采血孕周、尿素氮,尿酸,肌酐,β2-微球蛋白,α1-微球蛋白、乳酸脫氫酶無明顯相關(guān)性。6血清IFI16水平在預(yù)測(cè)子癇前期中的價(jià)值ROC曲線分析表明,以13.89ng/m L IFI16濃度作為預(yù)測(cè)子癇前期的臨界值時(shí),約登指數(shù)最大,其值為0.61,靈敏度和特異性分別為77.78%和83.33%,ROC曲線下面積為0.859;以15.6ng/m L IFI16水平作為區(qū)分輕度子癇前期患者和重度子癇前期患者的臨界值時(shí),約登指數(shù)最大,其值為0.38,靈敏度和特異性分別為90%和48%,ROC曲線下面積為0.658。結(jié)論IFI16可能通過損傷血管內(nèi)皮細(xì)胞參與了子癇前期的發(fā)病并有可能成為預(yù)測(cè)子癇前期發(fā)生的新的生物學(xué)指標(biāo)。
[Abstract]:Preeclampsia is one of the special diseases during pregnancy. It is characterized by persistent elevated blood pressure and proteinuria after 20 weeks of gestation. The incidence of preeclampsia is about 3%-8% worldwide, which seriously affects the health of mothers and infants. However, the exact pathogenesis of preeclampsia is still unclear. Human interferon-inducible protein 16 (IFI16) is a member of the HIN-200 family and mainly exists in hematopoietic cells, fibroblasts, epithelial cells, endothelial cells, trophoblasts and other cells. High levels of IFI16 in serum may be associated with endothelial cell injury. Oxidative stress, various inflammatory factors and cell density can trigger up-regulation of IFI16 expression. Considering that the placenta tissues of preeclampsia are in a state of strong oxidative stress and high inflammatory reaction, endothelial cell injury is the central link in the pathogenesis of endothelial cell injury. Objective To investigate the expression of IFI16 in placenta tissue and serum of normal and preeclampsia patients, and to detect the levels of IFI16 in serum, such as blood pressure, 24-hour urinary protein and serum total egg. The correlation between serum levels of IFI 16 and albumin, albumin, urea, uric acid, creatinine, cystatin C, beta 2-microglobulin, alpha 1-microglobulin, lactate dehydrogenase and endothelin-1 (ET-1) was analyzed. Finally, the receiver operator characteristic curve (ROC curve) and Jordan index were used to predict the serum IFI 16 levels. Methods The placenta tissues and serum of normal and preeclampsia pregnant women were collected and detected by immunohistochemistry, real-time fluorescence quantitative PCR and Western blotting respectively. Expressions of serum total protein, albumin, urea, uric acid, creatinine, cystatin C, beta 2-microglobulin, alpha 1-microglobulin, lactate dehydrogenase and 24-hour urinary protein were detected by automatic biochemical analyzer; serum levels of IFI 16 and ET-1 were detected by ELISA, and the relationship between serum IFI 16 levels and each index was analyzed. Correlation analysis. SPSS 21.0 was used to analyze and process the data. The results were expressed by mean (?) x (?) s) standard deviation. The differences between the two groups were compared by independent sample t test or chi-square test (2); Pearson correlation test was used for correlation analysis; ROC curve was used to judge whether serum IFI 16 level could be a new biological index. Results 1 Comparing the general clinical data and detection indexes of two groups of pregnant women 1.1 Comparing the general clinical data of two groups of pregnant women, the age of pregnant women in the preeclampsia group and the control group were (30.9 + 4.6) years old and (29.2 + 4.2) years old respectively, and the gestational weeks were (34.2). The gestational weeks of delivery were (35.3 3 2.6 2.6) and (38.6 1.0) weeks respectively. The systolic blood pressure were (156.6 6 [(9.6) mm Hg and (115.1 [(9.5) mm Hg, the diastolic blood pressure were (103.9 [(7.9 [(7.6) mm Hg] mm Hg and (73.6 [(7.6 [(5.9) mm Hg] Hg, the birth weight were (2170.3.3 [309.3 9.3 9.2 2 2) g and (3316.1 1 456.1 1 1 1 1 [(45 16.16.16.1)] mm Hg, respectively. The diastostolicblood pressure were (103.110.9 mg There was no significant difference between the two groups in age and gestational weeks (P 0.05). Systolic blood pressure, diastolic blood pressure and 24-hour urinary protein were higher in the preeclampsia group than in the control group, the difference was statistically significant (P 0.05). The preeclampsia group had lower gestational weeks and fetal weight than the control group. Statistical significance (P 0.05). 1. 17.4 [(94.4 [(94.5) and (263.0 [(66.8) micromol/L, creatinine concentrations were (53.6 [(11.5 [(53.6 [(11.5) and (44.6 [(6 6.6 [(44.6 [(6 6 6.6)) micromol/L/L), bet2-microglobulin concentrations were (2.3 [(0.6) mg/L and (1.8 [(0.0.4) mg/L, alpha1-microglobulin concentrations were (27.5 [(7.7.7.7) mg/L and (24.7 [(24.7 4.1) mg/L), cystatin C concentrations were (1.6 [(1.(1.1 +0.2) micromol/L, milk The serum levels of urea nitrogen, uric acid, creatinine, beta-2-microglobulin, alpha-1-microglobulin, cystatin C and lactate dehydrogenase in preeclampsia group were higher than those in control group (P 0.05). The levels of serum total protein and albumin in preeclampsia group were significantly higher than those in control group (P 0.05). The expression and localization of IFI16 protein in placenta tissues of both groups were significantly lower than those of the control group (P 0.05). The positive expression rate of IFI16 in placenta of normal group and preeclampsia group was 43% and 78%, respectively. The difference was statistically significant (_2 = 9.671, P 0.05). 3 The relative expression of IFI16 m RNA and protein in placenta of preeclampsia group was (1.65 + 0.08). The relative expression level of IFI 16 protein in placenta of preeclampsia group was also higher than that of control group (0.56.03). The difference was statistically significant (P 0.01). 4 The concentration of IFI 16 and ET-1 in serum of preeclampsia group was significantly higher than that of control group (P 0.01). The serum level of ET-1 in preeclampsia group was (41.6 [9.3] ng / L, and that in control group was (11.0 [4.9] ng / L). There was a significant difference between the two groups (P 0.01). The serum level of IFI-16 in preeclampsia group was (41.6 [9.3] ng / L] and that in preeclampsia group was (11.0 [4.9] ng / L]. There was a significant difference between the two groups (P 0.01). Sexual Pearson correlation analysis showed that serum IFI 16 levels were positively correlated with systolic blood pressure (r = 0.639, P 0.01), diastolic blood pressure (r = 0.514, P 0.01), 24-hour urinary protein (r = 0.707, P 0.01), cystatin C (r = 0.347, P 0.05) and serum ET-1 (r = 0.616, P 0.01), and negatively correlated with serum albumin (r =-0.366, P 0.05). Age, gestational age, blood urea nitrogen, uric acid, creatinine, beta-2-microglobulin, alpha-1-microglobulin, lactate dehydrogenase were not significantly correlated. The ROC curve analysis of serum IFI-16 level in predicting preeclampsia showed that when the concentration of 13.89ng/m L IFI-16 was used as the critical value for predicting preeclampsia, the Yorden index was the largest, with a value of 0.61, sensitivity and specificity. The heterosexuality was 77.78% and 83.33% respectively, and the area under ROC curve was 0.859. When the level of 15.6 ng/ml IFI 16 was used as the critical value to distinguish mild and severe pre-eclampsia patients, the Yorden index was the largest, with a value of 0.38, a sensitivity and specificity of 90% and 48%, respectively. The area under ROC curve was 0.658. Endothelial cells are involved in the pathogenesis of preeclampsia and may become a new biological indicator for predicting the occurrence of preeclampsia.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R714.244
，
本文編號(hào)：2216505
[Abstract]:Preeclampsia is one of the special diseases during pregnancy. It is characterized by persistent elevated blood pressure and proteinuria after 20 weeks of gestation. The incidence of preeclampsia is about 3%-8% worldwide, which seriously affects the health of mothers and infants. However, the exact pathogenesis of preeclampsia is still unclear. Human interferon-inducible protein 16 (IFI16) is a member of the HIN-200 family and mainly exists in hematopoietic cells, fibroblasts, epithelial cells, endothelial cells, trophoblasts and other cells. High levels of IFI16 in serum may be associated with endothelial cell injury. Oxidative stress, various inflammatory factors and cell density can trigger up-regulation of IFI16 expression. Considering that the placenta tissues of preeclampsia are in a state of strong oxidative stress and high inflammatory reaction, endothelial cell injury is the central link in the pathogenesis of endothelial cell injury. Objective To investigate the expression of IFI16 in placenta tissue and serum of normal and preeclampsia patients, and to detect the levels of IFI16 in serum, such as blood pressure, 24-hour urinary protein and serum total egg. The correlation between serum levels of IFI 16 and albumin, albumin, urea, uric acid, creatinine, cystatin C, beta 2-microglobulin, alpha 1-microglobulin, lactate dehydrogenase and endothelin-1 (ET-1) was analyzed. Finally, the receiver operator characteristic curve (ROC curve) and Jordan index were used to predict the serum IFI 16 levels. Methods The placenta tissues and serum of normal and preeclampsia pregnant women were collected and detected by immunohistochemistry, real-time fluorescence quantitative PCR and Western blotting respectively. Expressions of serum total protein, albumin, urea, uric acid, creatinine, cystatin C, beta 2-microglobulin, alpha 1-microglobulin, lactate dehydrogenase and 24-hour urinary protein were detected by automatic biochemical analyzer; serum levels of IFI 16 and ET-1 were detected by ELISA, and the relationship between serum IFI 16 levels and each index was analyzed. Correlation analysis. SPSS 21.0 was used to analyze and process the data. The results were expressed by mean (?) x (?) s) standard deviation. The differences between the two groups were compared by independent sample t test or chi-square test (2); Pearson correlation test was used for correlation analysis; ROC curve was used to judge whether serum IFI 16 level could be a new biological index. Results 1 Comparing the general clinical data and detection indexes of two groups of pregnant women 1.1 Comparing the general clinical data of two groups of pregnant women, the age of pregnant women in the preeclampsia group and the control group were (30.9 + 4.6) years old and (29.2 + 4.2) years old respectively, and the gestational weeks were (34.2). The gestational weeks of delivery were (35.3 3 2.6 2.6) and (38.6 1.0) weeks respectively. The systolic blood pressure were (156.6 6 [(9.6) mm Hg and (115.1 [(9.5) mm Hg, the diastolic blood pressure were (103.9 [(7.9 [(7.6) mm Hg] mm Hg and (73.6 [(7.6 [(5.9) mm Hg] Hg, the birth weight were (2170.3.3 [309.3 9.3 9.2 2 2) g and (3316.1 1 456.1 1 1 1 1 [(45 16.16.16.1)] mm Hg, respectively. The diastostolicblood pressure were (103.110.9 mg There was no significant difference between the two groups in age and gestational weeks (P 0.05). Systolic blood pressure, diastolic blood pressure and 24-hour urinary protein were higher in the preeclampsia group than in the control group, the difference was statistically significant (P 0.05). The preeclampsia group had lower gestational weeks and fetal weight than the control group. Statistical significance (P 0.05). 1. 17.4 [(94.4 [(94.5) and (263.0 [(66.8) micromol/L, creatinine concentrations were (53.6 [(11.5 [(53.6 [(11.5) and (44.6 [(6 6.6 [(44.6 [(6 6 6.6)) micromol/L/L), bet2-microglobulin concentrations were (2.3 [(0.6) mg/L and (1.8 [(0.0.4) mg/L, alpha1-microglobulin concentrations were (27.5 [(7.7.7.7) mg/L and (24.7 [(24.7 4.1) mg/L), cystatin C concentrations were (1.6 [(1.(1.1 +0.2) micromol/L, milk The serum levels of urea nitrogen, uric acid, creatinine, beta-2-microglobulin, alpha-1-microglobulin, cystatin C and lactate dehydrogenase in preeclampsia group were higher than those in control group (P 0.05). The levels of serum total protein and albumin in preeclampsia group were significantly higher than those in control group (P 0.05). The expression and localization of IFI16 protein in placenta tissues of both groups were significantly lower than those of the control group (P 0.05). The positive expression rate of IFI16 in placenta of normal group and preeclampsia group was 43% and 78%, respectively. The difference was statistically significant (_2 = 9.671, P 0.05). 3 The relative expression of IFI16 m RNA and protein in placenta of preeclampsia group was (1.65 + 0.08). The relative expression level of IFI 16 protein in placenta of preeclampsia group was also higher than that of control group (0.56.03). The difference was statistically significant (P 0.01). 4 The concentration of IFI 16 and ET-1 in serum of preeclampsia group was significantly higher than that of control group (P 0.01). The serum level of ET-1 in preeclampsia group was (41.6 [9.3] ng / L, and that in control group was (11.0 [4.9] ng / L). There was a significant difference between the two groups (P 0.01). The serum level of IFI-16 in preeclampsia group was (41.6 [9.3] ng / L] and that in preeclampsia group was (11.0 [4.9] ng / L]. There was a significant difference between the two groups (P 0.01). Sexual Pearson correlation analysis showed that serum IFI 16 levels were positively correlated with systolic blood pressure (r = 0.639, P 0.01), diastolic blood pressure (r = 0.514, P 0.01), 24-hour urinary protein (r = 0.707, P 0.01), cystatin C (r = 0.347, P 0.05) and serum ET-1 (r = 0.616, P 0.01), and negatively correlated with serum albumin (r =-0.366, P 0.05). Age, gestational age, blood urea nitrogen, uric acid, creatinine, beta-2-microglobulin, alpha-1-microglobulin, lactate dehydrogenase were not significantly correlated. The ROC curve analysis of serum IFI-16 level in predicting preeclampsia showed that when the concentration of 13.89ng/m L IFI-16 was used as the critical value for predicting preeclampsia, the Yorden index was the largest, with a value of 0.61, sensitivity and specificity. The heterosexuality was 77.78% and 83.33% respectively, and the area under ROC curve was 0.859. When the level of 15.6 ng/ml IFI 16 was used as the critical value to distinguish mild and severe pre-eclampsia patients, the Yorden index was the largest, with a value of 0.38, a sensitivity and specificity of 90% and 48%, respectively. The area under ROC curve was 0.658. Endothelial cells are involved in the pathogenesis of preeclampsia and may become a new biological indicator for predicting the occurrence of preeclampsia.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R714.244
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本文編號(hào)：2216505
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