miRNA表達(dá)譜的變化作為2型糖尿病早期診斷標(biāo)志物的研究
發(fā)布時(shí)間：2019-06-11 00:34
【摘要】：目的:T2DM已經(jīng)成為影響全球健康的元兇之一,目前尚無特異性T2DM早期預(yù)測標(biāo)志物,因此尋找其早期預(yù)測生物標(biāo)志物十分必要。本文旨在篩選不同糖耐量狀態(tài)下micro RNA的表達(dá)差異,尋找T2DM潛在的生物標(biāo)志物,并初步探討其在T2DM中的可能作用,為早期預(yù)測和干預(yù)T2DM提供新的靶點(diǎn)。方法:1.選取健康對照者,糖調(diào)節(jié)受損者,2型糖尿病者各2例,應(yīng)用micro RNA基因芯片方法篩選各組間差異表達(dá)的mi RNA,篩選標(biāo)準(zhǔn)為兩組間比較有顯著性差異(倍數(shù)差異2或倍數(shù)差異0.5),隨后在得到的有顯著性差異的mi RNA中選取表達(dá)量高,倍數(shù)差異明顯的mi RNA進(jìn)行進(jìn)一步驗(yàn)證及研究。2.選取健康對照者108例,糖調(diào)節(jié)受損者92例,2型糖尿病者117例,分別抽取其清晨空腹外周靜脈血,提取血清,應(yīng)用實(shí)時(shí)熒光定量PCR技術(shù)檢測三組中mi R-3200-5p、mi R-100-5p、mi R-574-5p、mi R-3135b、mi R-1972、mi R-133b、mi R-1273f表達(dá)量以驗(yàn)證芯片的準(zhǔn)確性,分析7個(gè)mi RNA與臨床參數(shù)的關(guān)系,并建立ROC曲線,評價(jià)其診斷的靈敏性和特異性。最后用靶基因預(yù)測軟件和KEGG通路分析軟件,分析這7個(gè)mi RNA在糖尿病中可能發(fā)揮的作用。結(jié)果：1.micro RNA基因芯片結(jié)果顯示,三組共2549個(gè)mi RNA,以倍數(shù)差異2或倍數(shù)差異0.5為標(biāo)準(zhǔn),健康對照組與糖調(diào)節(jié)受損組間有5個(gè)差異表達(dá)的mi RNA,其中上調(diào)5個(gè),下調(diào)0個(gè);糖調(diào)節(jié)受損組與2型糖尿病組間有4個(gè)差異表達(dá)的mi RNA,其中上調(diào)2個(gè),下調(diào)2個(gè);健康對照組和2型糖尿病組有26個(gè)差異表達(dá)的mi RNA,其中上調(diào)17個(gè),下調(diào)9個(gè)。2.根據(jù)這些mi RNA的組間差異,表達(dá)量及相關(guān)的生物學(xué)分析,選取7個(gè)mi RNA(mi R-3200-5p、mi R-100-5p、mi R-574-5p、mi R-3135b、mi R-1972、mi R-133b、mi R-1273f)進(jìn)行進(jìn)一步驗(yàn)證和研究。q RT-PCR技術(shù)檢測結(jié)果顯示,mi R-3200-5p、mi R-574-5p、mi R-3135b、mi R-100-5p和mi R-1972在健康對照組、糖調(diào)節(jié)受損組和2型糖尿病組的表達(dá)依次降低,其中mi R-3200-5p和mi R-574-5p三組間均有顯著性差異(P0.05),mi R-100-5p在健康對照組和糖調(diào)節(jié)受損組之間無統(tǒng)計(jì)學(xué)差異(P0.05),mi R-1972在糖調(diào)節(jié)受損組和2型糖尿病組間無統(tǒng)計(jì)學(xué)差異(P0.05)。mi R-133b、mi R-1273f在健康對照組、糖調(diào)節(jié)受損組和2型糖尿病組的表達(dá)先降低再上升,但mi R-133b在三組間均有統(tǒng)計(jì)學(xué)差異(P0.05),mi R-1273f在糖調(diào)節(jié)受損和2型糖尿病組間無統(tǒng)計(jì)學(xué)差異(P0.05)。其中,mi R-100-5p的PCR驗(yàn)證結(jié)果與基因芯片結(jié)果相反,其余6個(gè)mi RNA的PCR驗(yàn)證結(jié)果與基因芯片結(jié)果一致。3.對7個(gè)mi RNA不同組合來評估其在早期診斷2型糖尿病中的價(jià)值,發(fā)現(xiàn)mi R-574-5p、mi R-133b、mi RNA-3135b、mi R-1273f、mi R-1972聯(lián)合檢驗(yàn)時(shí),區(qū)分健康對照組與糖調(diào)節(jié)受損組的診斷準(zhǔn)確性最高,其曲線下面積為0.805(P0.01);mi R-3200-5p、mi R-574-5p、mi R-133b、mi R-100-5p聯(lián)合檢驗(yàn)時(shí),區(qū)分糖調(diào)節(jié)受損組與2型糖尿病組的診斷準(zhǔn)確性最高,其曲線下面積為0.738(P0.01);mi R-3200-5p、mi R-574-5p、mi R-1273f、mi R-100-5p聯(lián)合檢驗(yàn)時(shí),區(qū)分健康對照組與2型糖尿病組的診斷準(zhǔn)確性最大,其曲線下面積為0.935(P0.01)。多元相關(guān)分析顯示,與健康對照組相比,糖調(diào)節(jié)受損組和2型糖尿病組的FBG、TG、Hb A1c、BMI、WHR顯著升高(P0.05),HDL-C顯著降低(P0.05);與糖調(diào)節(jié)受損組相比,2型糖尿病組FBG、SBP、DBP、TG、Hb A1c、WHR、BMI顯著升高(P0.05),HDL-C顯著降低(P0.05)。結(jié)論：1.mi RNA基因芯片結(jié)果顯示,當(dāng)倍數(shù)差異2或倍數(shù)差異0.5時(shí),三組間分別有不同差異表達(dá)的mi RNA;2.篩選出7個(gè)mi RNA(分別為mi R-3200-5p、mi R-100-5p、mi R-574-5p、mi R-3135b、mi R-1972、mi R-133b、mi R-1273f)進(jìn)行q RT-PCR驗(yàn)證,發(fā)現(xiàn)其中的6個(gè)mi RNA(除mi R-100-5p)在糖尿病的進(jìn)程中有顯著變化趨勢,有望成為2型糖尿病早期生物學(xué)標(biāo)志物。3.7種mi RNA與臨床資料密切相關(guān),且在不同的組合情況下的診斷價(jià)值不同,聯(lián)合檢測可提高診斷效率。同時(shí),7種mi RNA的靶基因可能通過蛋白消化和吸收、TGF-β信號通路、胰島分泌、p53信號通路、磷酸化信號系統(tǒng)等分子生物學(xué)作用和代謝通路發(fā)揮作用。
[Abstract]:Objective: T2DM has become one of the elements that affect the global health, and there is no specific early prediction marker of T2DM, so it is necessary to find its early prediction biomarkers. The purpose of this study is to screen the differential expression of micro RNA in different glucose tolerance states, to find the potential biomarkers of T2DM, and to explore its possible role in T2DM and to provide a new target for early prediction and intervention of T2DM. Method:1. The expression of mi-RNA in each group was selected by microRNA gene chip method, and the screening criteria were significant difference (multiple difference of 2 or multiple difference of 0.5). And then the mi RNA with high expression quantity and the multiple difference is selected to be further verified and studied in the obtained mi RNA with significant difference. 108 patients with healthy controls,92 patients with impaired glucose regulation and 117 patients with type 2 diabetes were selected, and the fasting peripheral venous blood in the morning was extracted, serum was extracted, and the mi R-3200-5p, mi R-100-5p, mi R-574-5p, mi R-3135b, mi R-1972, mi R-133b were detected by real-time fluorescence quantitative PCR. The expression of mi R-1273f was used to verify the accuracy of the chip, the relationship between the 7 mi RNA and the clinical parameters was analyzed, and the ROC curve was established to evaluate the sensitivity and specificity of the diagnosis. Finally, the target gene prediction software and the KEGG pathway analysis software were used to analyze the possible role of the seven mi-RNA in the diabetes. Results: The results of microRNA gene chip showed that there were a total of 2549 mi-RNA in the three groups, with a multiple difference of 2 or a multiple of 0.5 as the standard. There were 4 different expression of mi-RNA in the group with impaired glucose regulation group and type 2 diabetes group, of which 2 were up-regulated and 2 were down-regulated; there were 26 difference expression of mi-RNA in the healthy control group and type 2 diabetes group, including 17 up-regulation and 9 down-regulation. 7 mi-RNA (mi R-3200-5p, mi R-100-5p, mi R-574-5p, mi R-3135b, mi R-1972, mi R-133b, mi R-1273f) were selected for further verification and study according to the inter-group difference, expression and related biological analysis of these mi-RNA. The results of q-RT-PCR showed that the expression of mi R-3200-5p, mi R-574-5p, mi R-3135b, mi R-100-5p, and mi R-1972 in healthy control group, sugar-adjusted damaged group and type 2 diabetes group decreased in sequence, among which, there was a significant difference among the three groups of mi R-3200-5p and mi R-574-5p (P0.05). mi r-100-5p had no statistical difference between the healthy control group and the impaired glucose control group (P0.05). mi r-1972 no significant difference between the sugar-adjusted and 2-type diabetic group (p0.05). mi r-133b, mi r-1273f, in the healthy control group, the expression of the sugar-adjusted damaged group and the type 2 diabetes group was first reduced and then increased, But the mi R-133b had a statistical difference among the three groups (P0.05), and the mi R-1273f had no statistical difference between the sugar regulation and the type 2 diabetes group (P0.05). The results of PCR validation of mi R-100-5p were in contrast to the results of gene chip, and the results of PCR of the remaining six mi-RNA were consistent with the results of gene chip. The results showed that the diagnostic accuracy of mi R-574-5p, mi R-133b, mi-RNA-3135b, mi R-1273f, mi R-1972 was the highest in the early diagnosis of type 2 diabetes, the area of the curve was 0.805 (P0.01), mi R-3200-5p, mi R-574-5p, In the combined test of mi R-133b, mi R-100-5p, the diagnostic accuracy of the sugar-adjusted damaged group and type 2 diabetes group was the highest, the area under the curve was 0.738 (P0.01), mi R-3200-5p, mi R-574-5p, mi R-1273f, mi R-100-5p combined test, the diagnostic accuracy of the healthy control group and type 2 diabetes group was the largest, The area under the curve is 0.935 (P0.01). The multivariate correlation analysis showed that the FBG, TG, Hb, BMI, WHR of the damaged group and type 2 diabetes group were significantly higher than those in the healthy control group (P0.05), and the HDL-C was significantly lower (P0.05); and the FBG, SBP, DBP, TG, Hb, and WHR in type 2 diabetes group were compared with that of the group 2 diabetes. The BMI was significantly higher (P0.05), and the HDL-C was significantly lower (P0.05). Conclusion: The results of 1.mi RNA gene chip show that when the multiple difference is 2 or a multiple of 0.5, there are different expression of mi-RNA in the three groups. 7 mi-RNA (mi R-3200-5p, mi R-100-5p, mi R-574-5p, mi R-3135b, mi R-1972, mi R-133b, mi R-1273f) were screened for q-RT-PCR. It is expected to be an early biological marker of type 2 diabetes. 3.7 kinds of mi-RNA are closely related to clinical data, and in different combinations, the diagnostic value is different, and the combined detection can improve the diagnostic efficiency. At the same time, the target gene of seven kinds of mi-RNA can play a role in molecular biology and metabolic pathway of protein digestion and absorption, TGF-signaling pathway, pancreatic islet secretion, p53 signal pathway, and phosphorylation signal system.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R587.1
本文編號：2496856
[Abstract]:Objective: T2DM has become one of the elements that affect the global health, and there is no specific early prediction marker of T2DM, so it is necessary to find its early prediction biomarkers. The purpose of this study is to screen the differential expression of micro RNA in different glucose tolerance states, to find the potential biomarkers of T2DM, and to explore its possible role in T2DM and to provide a new target for early prediction and intervention of T2DM. Method:1. The expression of mi-RNA in each group was selected by microRNA gene chip method, and the screening criteria were significant difference (multiple difference of 2 or multiple difference of 0.5). And then the mi RNA with high expression quantity and the multiple difference is selected to be further verified and studied in the obtained mi RNA with significant difference. 108 patients with healthy controls,92 patients with impaired glucose regulation and 117 patients with type 2 diabetes were selected, and the fasting peripheral venous blood in the morning was extracted, serum was extracted, and the mi R-3200-5p, mi R-100-5p, mi R-574-5p, mi R-3135b, mi R-1972, mi R-133b were detected by real-time fluorescence quantitative PCR. The expression of mi R-1273f was used to verify the accuracy of the chip, the relationship between the 7 mi RNA and the clinical parameters was analyzed, and the ROC curve was established to evaluate the sensitivity and specificity of the diagnosis. Finally, the target gene prediction software and the KEGG pathway analysis software were used to analyze the possible role of the seven mi-RNA in the diabetes. Results: The results of microRNA gene chip showed that there were a total of 2549 mi-RNA in the three groups, with a multiple difference of 2 or a multiple of 0.5 as the standard. There were 4 different expression of mi-RNA in the group with impaired glucose regulation group and type 2 diabetes group, of which 2 were up-regulated and 2 were down-regulated; there were 26 difference expression of mi-RNA in the healthy control group and type 2 diabetes group, including 17 up-regulation and 9 down-regulation. 7 mi-RNA (mi R-3200-5p, mi R-100-5p, mi R-574-5p, mi R-3135b, mi R-1972, mi R-133b, mi R-1273f) were selected for further verification and study according to the inter-group difference, expression and related biological analysis of these mi-RNA. The results of q-RT-PCR showed that the expression of mi R-3200-5p, mi R-574-5p, mi R-3135b, mi R-100-5p, and mi R-1972 in healthy control group, sugar-adjusted damaged group and type 2 diabetes group decreased in sequence, among which, there was a significant difference among the three groups of mi R-3200-5p and mi R-574-5p (P0.05). mi r-100-5p had no statistical difference between the healthy control group and the impaired glucose control group (P0.05). mi r-1972 no significant difference between the sugar-adjusted and 2-type diabetic group (p0.05). mi r-133b, mi r-1273f, in the healthy control group, the expression of the sugar-adjusted damaged group and the type 2 diabetes group was first reduced and then increased, But the mi R-133b had a statistical difference among the three groups (P0.05), and the mi R-1273f had no statistical difference between the sugar regulation and the type 2 diabetes group (P0.05). The results of PCR validation of mi R-100-5p were in contrast to the results of gene chip, and the results of PCR of the remaining six mi-RNA were consistent with the results of gene chip. The results showed that the diagnostic accuracy of mi R-574-5p, mi R-133b, mi-RNA-3135b, mi R-1273f, mi R-1972 was the highest in the early diagnosis of type 2 diabetes, the area of the curve was 0.805 (P0.01), mi R-3200-5p, mi R-574-5p, In the combined test of mi R-133b, mi R-100-5p, the diagnostic accuracy of the sugar-adjusted damaged group and type 2 diabetes group was the highest, the area under the curve was 0.738 (P0.01), mi R-3200-5p, mi R-574-5p, mi R-1273f, mi R-100-5p combined test, the diagnostic accuracy of the healthy control group and type 2 diabetes group was the largest, The area under the curve is 0.935 (P0.01). The multivariate correlation analysis showed that the FBG, TG, Hb, BMI, WHR of the damaged group and type 2 diabetes group were significantly higher than those in the healthy control group (P0.05), and the HDL-C was significantly lower (P0.05); and the FBG, SBP, DBP, TG, Hb, and WHR in type 2 diabetes group were compared with that of the group 2 diabetes. The BMI was significantly higher (P0.05), and the HDL-C was significantly lower (P0.05). Conclusion: The results of 1.mi RNA gene chip show that when the multiple difference is 2 or a multiple of 0.5, there are different expression of mi-RNA in the three groups. 7 mi-RNA (mi R-3200-5p, mi R-100-5p, mi R-574-5p, mi R-3135b, mi R-1972, mi R-133b, mi R-1273f) were screened for q-RT-PCR. It is expected to be an early biological marker of type 2 diabetes. 3.7 kinds of mi-RNA are closely related to clinical data, and in different combinations, the diagnostic value is different, and the combined detection can improve the diagnostic efficiency. At the same time, the target gene of seven kinds of mi-RNA can play a role in molecular biology and metabolic pathway of protein digestion and absorption, TGF-signaling pathway, pancreatic islet secretion, p53 signal pathway, and phosphorylation signal system.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R587.1
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 Mustafa Abdo Saif Dehwah;;MicroRNAs and Type 2 Diabetes/Obesity[J];遺傳學(xué)報(bào);2012年01期
，本文編號：2496856
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