IFI16在子痫前期胎盘组织和血清中的表达及与子痫前期发病的相关性
发布时间：2018-09-01 07:21
【摘要】：子痫前期是妊娠期特有的疾病之一,以妊娠20周以后新发的持续性血压升高并伴有明显蛋白尿为主要特征,在全世界的发病率约为3%~8%,严重影响母婴健康。然而到目前为止,子痫前期的确切发病机制仍不清楚。现在研究认为,内皮细胞损伤是子痫前期发病的中心环节。人γ干扰素诱导蛋白16(Interferon-inducible protein 16,IFI16)属于HIN-200家族成员,主要存在于造血细胞、成纤维细胞、上皮细胞、内皮细胞、滋养细胞等细胞内。最近研究发现,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可能与内皮细胞损伤有关。氧化应激、各种炎性因子和细胞密度可触发IFI16表达上调。考虑到子痫前期胎盘组织处于强氧化应激和高炎性反应状态,内皮细胞损伤是其发病的中心环节,推测IFI16蛋白可能与子痫前期发病有关。然而,目前关于IFI16蛋白在子痫前期中的研究尚比较少见。目的本研究通过检测IFI16在正常组和子痫前期组胎盘组织和血清中的表达,并对血清IFI16水平与内皮细胞损伤相关的检测指标,如血压、24h尿蛋白、血清总蛋白、白蛋白、尿素、尿酸、肌酐、胱抑素C、β2-微球蛋白、α1-微球蛋白、乳酸脱氢酶和内皮素-1(Endothelin-1,ET-1)等指标之间的相关性进行分析,最后利用受试者工作曲线(receiver operator characteristic curve,ROC曲线)及约登指数评判血清IFI16水平在预测子痫前期中的价值,从而探讨IFI16与子痫前期发病的相关性,并为该指标的临床应用提供实验室依据。方法收集正常组和子痫前期组孕妇胎盘组织和血清,分别采用免疫组化法、实时荧光定量PCR技术和蛋白印迹法检测胎盘组织中IFI16的表达;采用全自动生化分析仪检测两组孕妇血清总蛋白、白蛋白、尿素、尿酸、肌酐、胱抑素C、β2-微球蛋白、α1-微球蛋白、乳酸脱氢酶及24h尿蛋白等生化指标;采用ELISA方法检测血清中IFI16和ET-1的水平,并对血清IFI16水平与各检测指标之间的相关性进行分析。应用SPSS 21.0进行数据的分析和处理,结果以均数±标准差(?x±s)表示,两组间差异性比较采用独立样本t检验或卡方检验(χ2);相关性分析采用Pearson相关性检验;ROC曲线用于评判血清IFI16水平能否作为一种新的生物学指标用于预测子痫前期的发生,并确定相应的预测界值。以α=0.05为检验水准。结果1两组孕妇一般临床资料和检测指标的比较1.1两组孕妇一般临床资料的比较子痫前期组和对照组孕妇年龄分别为(30.9±4.6)岁和(29.2±4.2)岁,采血孕周分别为(34.2±2.3)周和(34.3±1.0)周,分娩孕周分别为(35.3±2.6)周和(38.6±1.0)周,收缩压分别为(156.6±9.6)mm Hg和(115.1±9.5)mm Hg,舒张压分别为(103.9±7.6)mm Hg和(73.6±5.9)mm Hg,新生儿体重分别为(2170.3±309.2)g和(3316.1±456.1)g,24小时尿蛋白量分别为(2003.5±110.9)mg和(83.2±17.2)mg。经统计学分析,两组孕妇年龄和采血孕周相比,差异无统计学意义(P0.05);子痫前期组收缩压、舒张压以及24小时尿蛋白定量均高于对照组,差异具有统计学意义(P0.05);子痫前期组分娩孕周和胎儿体重均低于对照组,差异具有统计学意义(P0.05)。1.2两组孕妇临床检测指标的比较子痫前期组和对照组孕妇血清总蛋白浓度分别为(56.3±6.8)g/L和(61.6±5.5)g/L,白蛋白浓度分别为(29.8±4.2)g/L和(34.7±2.9)g/L,尿素氮浓度分别为(4.0±0.8)mmol/L和(3.5±0.7)mmol/L,尿酸浓度分别为(317.4±94.5)μmol/L和(263.0±66.8)μmol/L,肌酐浓度分别为(53.6±11.5)μmol/L和(44.6±6.6)μmol/L,β2-微球蛋白浓度分别为(2.3±0.6)mg/L和(1.8±0.4)mg/L,α1-微球蛋白浓度分别为(27.5±7.7)mg/L和(24.7±4.1)mg/L,胱抑素C浓度分别为(1.6±0.4)μmol/L和(1.1±0.2)μmol/L,乳酸脱氢酶浓度分别为(256.6±61.8)U/L和(198.5±30.1)U/L。经统计学分析,子痫前期组孕妇血清尿素氮、尿酸、肌酐、β2-微球蛋白、α1-微球蛋白、胱抑素C、乳酸脱氢酶水平均高于对照组,差异具有统计学意义(P0.05);子痫前期组孕妇血清总蛋白和白蛋白水平均低于对照组,差异具有统计学意义(P0.05)。2两组孕妇胎盘组织中IFI16蛋白的表达和定位两组孕妇胎盘滋养细胞和血管内皮细胞的细胞质中均有IFI16蛋白的表达。与对照组相比,IFI16蛋白在子痫前期组胎盘组织中的阳性表达率明显增高(IFI16在正常组和子痫前期组胎盘组织中的阳性表达率分别为43%和78%),两组比较,差异具有统计学意义(χ2=9.671,P0.05)。3两组孕妇胎盘组织中IFI16 m RNA和蛋白质表达水平的比较子痫前期组胎盘组织中IFI16 m RNA的相对表达量(1.65±0.08)高于对照(0.72±0.05),两组比较,差异具有统计学意义(P0.01);子痫前期组胎盘组织中IFI16蛋白的相对表达水平(1.58±0.06)亦高于对照组(0.56±0.03),差异均具有统计学意义(P0.01)。4两组孕妇血清中IFI16及ET-1水平的比较子痫前期组血清IFI16的浓度为(20.68±8.9)ng/m L,对照组为(10.08±4.1)ng/m L,两组比较,差异有统计学意义(P0.01);子痫前期组血清ET-1的水平为(41.6±9.3)ng/L,对照组为(11.0±4.9)ng/L,两组比较,差异有统计学意义(P0.01)。5子痫前期组孕妇血清IFI16水平与各检测指标之间的相关性Pearson相关性分析表明,子痫前期组孕妇血清IFI16水平与孕妇收缩压(r=0.639,P0.01)、舒张压(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)呈正相关,与血清白蛋白水平呈负相关(r=-0.366,P0.05);与孕妇年龄、采血孕周、尿素氮,尿酸,肌酐,β2-微球蛋白,α1-微球蛋白、乳酸脱氢酶无明显相关性。6血清IFI16水平在预测子痫前期中的价值ROC曲线分析表明,以13.89ng/m L IFI16浓度作为预测子痫前期的临界值时,约登指数最大,其值为0.61,灵敏度和特异性分别为77.78%和83.33%,ROC曲线下面积为0.859;以15.6ng/m L IFI16水平作为区分轻度子痫前期患者和重度子痫前期患者的临界值时,约登指数最大,其值为0.38,灵敏度和特异性分别为90%和48%,ROC曲线下面积为0.658。结论IFI16可能通过损伤血管内皮细胞参与了子痫前期的发病并有可能成为预测子痫前期发生的新的生物学指标。
[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.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.244
，
本文编号：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.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.244
，
本文编号：2216505
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