circRNA-ZNF532、circRNA-HIPK3与糖尿病视网膜病变的相关性研究

doi:10.11958/20230048

摘要/Abstract

摘要：

目的探讨环状RNA（circRNA）-锌指蛋白532（ZNF532）、circRNA-同源域相互作用蛋白激酶3（HIPK3）与糖尿病视网膜病变（DR）的相关性。方法纳入109例DR患者（DR组）、110例单纯糖尿病患者（DM组）和65例健康体检志愿者（对照组）。实时荧光定量聚合酶链反应（qPCR）检测血清circRNA-ZNF532、circRNA-HIPK3表达，酶联免疫吸附试验检测血清血管内皮生长因子（VEGF）、白细胞介素（IL）-1β、肿瘤坏死因子（TNF）-α、IL-6水平。Pearson分析DR患者血清circRNA-ZNF532、circRNA-HIPK3表达与血清VEGF、IL-1β、TNF-α、IL-6水平相关性。单因素和多因素Logistic回归分析DR危险因素，受试者工作特征（ROC）曲线分析circRNA-ZNF532、circRNA-HIPK3诊断DR的价值。结果 DR组血清circRNA-ZNF532、circRNA-HIPK3表达以及VEGF、IL-1β、TNF-α、IL-6水平高于DM组和对照组（P<0.05）。DR组血清circRNA-ZNF532、circRNA-HIPK3表达与VEGF、IL-1β、TNF-α、IL-6水平均呈正相关（P<0.05）。多因素Logistic回归分析结果显示高水平circRNA-ZNF532、circRNA-HIPK3及VEGF是糖尿病患者发生DR的独立危险因素（P<0.05）。三者诊断糖尿病患者发生DR的曲线下面积分别为0.736、0.740和0.864，联合诊断高于单独诊断（P<0.05）。结论糖尿病患者血清中circRNA-ZNF532、circRNA-HIPK3表达上调与DR发生有关，两者可以作为DR辅助诊断的潜在指标。

关键词: 糖尿病视网膜病变, RNA, 环状, 转录激活因子类, tristetraprolin蛋白, 同源盒结构域蛋白质类, 炎症, 血管内皮生长因子类, 同源域相互作用蛋白激酶3

Abstract:

Objective To investigate the correlation between circrNa-Zinc finger protein 532 (ZNF532), circrNa-homologous domain interaction protein kinase-3 (HIPK3) and diabetic retinopathy (DR). Methods A total of 109 patients with DR (DR group), 110 patients with simple diabetes (DM group) and 65 healthy volunteers (control group) were selected from our hospital from October 2020 to October 2022. Serum circRNA-ZNF532 and circRNA-HIPK3 expression levels were detected by real-time quantitative polymerase chain reaction (RT-PCR), and serum vascular endothelial growth factor (VEGF), interleukin (IL) -1β, tumor necrosis factor (TNF) -α and IL-6 levels were detected by enzyme-linked immunosoradsorption assay. The correlation between serum circRNA-ZNF532 and circRNA-HIPK3 expression and serum levels of VEGF, IL-1β, TNF-α and IL-6 were analyzed by Pearson method. Univariate Logistic regression analysis and multivariate Logistic regression analysis were used for DR risk factors, and receiver operating characteristic curve (ROC) was used for analysis of circRNA-ZNF532 and circRNA-HIPK3 diagnostic value of DR. Results Serum expressions of circRNA-ZNF532 and circRNA-HIPK3 and levels of VEGF, IL-1β, TNF-α and IL-6 were significantly higher in the DR group than those in the DM group and the control group (P<0.05). Serum circRNA-ZNF532 and circRNA-HIPK3 expression were positively correlated with levels of VEGF, IL-1β, TNF-α and IL-6 in the DR group (P<0.05). Multivariate Logistic regression analysis showed that high levels of VEGF, circRNA-ZNF532 and circRNA-HIPK3 were risk factors for developing DR in diabetic patients (P<0.05). The areas under the curve of circRNA-ZNF532, circRNA-HIPK3 and VEGF in the diagnosis of DR in diabetic patients were 0.736, 0.740 and 0.864. Combined diagnosis was higher than that of circRNA-ZNF532, circRNA-HIPK3 and VEGF alone (P<0.05). Conclusion The up-regulated expression levels of circRNA-ZNF532 and circRNA-HIPK3 in serum of diabetic patients are related to the occurrence of DR, and both circRNA-ZNF532 and circRNA-HIPK3 can be used as potential indicators for the auxiliary diagnosis of DR.

Key words: diabetic retinopathy, RNA, circular, activating transcription factors, tristetraprolin, homeodomain proteins, inflammation, vascular endothelial growth factors, circrNa-homologous domain interaction protein kinase-3

中图分类号:

R774.13

图/表 6

参考文献 18

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组别	n	年龄/岁		男性		BMI/（kg/m²）		吸烟史		饮酒史		糖尿病病程/年	高血压	高脂血症		DR		糖尿病周围神经病变		冠心病
对照组	65	58.76±9.62		39（60.0）		22.90±1.53		31（47.69）		26（40.0）		-	-	-		-		-		-
DM组	110	58.51±9.27		60（54.5）		22.91±1.62		63（57.27）		54（49.1）		4.91±1.32	50（45.5）	53（48.2）		21（19.1）		17（15.5）		23（20.9）
DR组	109	60.54±8.78		62（56.9）		23.05±1.79		62（56.88）		51（46.8）		7.19±2.21	71（65.1）	50（45.9）		26（23.9）		18（16.5）		21（19.3）
F、χ²或t		1.518		0.498		0.236		1.787		1.493		9.270^＊＊	8.579^＊＊	0.117		0.737		0.046		0.092
组别	脑卒中		降糖治疗				TG/ （mmol/L）		TC/ （mmol/L）		Cr/ （μmol/L）		FPG/（mmol/L）		FINS/（mmol/L）		HOMA-IR		HbA1c/%
组别	脑卒中		口服降糖药		联合胰岛素		TG/ （mmol/L）		TC/ （mmol/L）		Cr/ （μmol/L）		FPG/（mmol/L）		FINS/（mmol/L）		HOMA-IR		HbA1c/%
对照组	-		-		-		1.46±0.16		4.96±0.60		62.72±5.72		4.39±0.79		4.13±1.06		1.27±0.32		5.22±0.70
DM组	15（13.6）		65（59.1）		45（40.9）		1.89±0.25^a		5.55±0.46^a		87.81±11.05^a		7.48±1.04^a		10.11±2.05^a		3.24±0.41^a		7.69±1.09^a
DR组	17（15.6）		60（55.1）		49（44.9）		1.89±0.27^a		5.65±0.39^a		90.73±9.24^a		8.97±1.32^ab		12.38±2.41^ab		4.88±0.57^ab		9.05±0.96^ab
F、χ²或t	0.169		0.366				87.260^＊＊		47.176^＊＊		204.144^＊＊		351.408^＊＊		348.849^＊＊		1241.548^＊＊		320.678^＊＊

组别	n	年龄/岁		男性		BMI/（kg/m²）		吸烟史		饮酒史		糖尿病病程/年	高血压	高脂血症		DR		糖尿病周围神经病变		冠心病
对照组	65	58.76±9.62		39（60.0）		22.90±1.53		31（47.69）		26（40.0）		-	-	-		-		-		-
DM组	110	58.51±9.27		60（54.5）		22.91±1.62		63（57.27）		54（49.1）		4.91±1.32	50（45.5）	53（48.2）		21（19.1）		17（15.5）		23（20.9）
DR组	109	60.54±8.78		62（56.9）		23.05±1.79		62（56.88）		51（46.8）		7.19±2.21	71（65.1）	50（45.9）		26（23.9）		18（16.5）		21（19.3）
F、χ²或t		1.518		0.498		0.236		1.787		1.493		9.270^＊＊	8.579^＊＊	0.117		0.737		0.046		0.092
组别	脑卒中		降糖治疗				TG/ （mmol/L）		TC/ （mmol/L）		Cr/ （μmol/L）		FPG/（mmol/L）		FINS/（mmol/L）		HOMA-IR		HbA1c/%
组别	脑卒中		口服降糖药		联合胰岛素		TG/ （mmol/L）		TC/ （mmol/L）		Cr/ （μmol/L）		FPG/（mmol/L）		FINS/（mmol/L）		HOMA-IR		HbA1c/%
对照组	-		-		-		1.46±0.16		4.96±0.60		62.72±5.72		4.39±0.79		4.13±1.06		1.27±0.32		5.22±0.70
DM组	15（13.6）		65（59.1）		45（40.9）		1.89±0.25^a		5.55±0.46^a		87.81±11.05^a		7.48±1.04^a		10.11±2.05^a		3.24±0.41^a		7.69±1.09^a
DR组	17（15.6）		60（55.1）		49（44.9）		1.89±0.27^a		5.65±0.39^a		90.73±9.24^a		8.97±1.32^ab		12.38±2.41^ab		4.88±0.57^ab		9.05±0.96^ab
F、χ²或t	0.169		0.366				87.260^＊＊		47.176^＊＊		204.144^＊＊		351.408^＊＊		348.849^＊＊		1241.548^＊＊		320.678^＊＊

组别	n	circRNA-ZNF532	circRNA-HIPK3	VEGF/（ng/L）	IL-1β/（ng/L）	TNF-α/（ng/L）	IL-6/（ng/L）
对照组	65	1.60±0.31	1.15±0.29	48.63±13.36	62.20±12.17	3.77±1.42	49.96±9.80
DM组	110	1.65±0.34	1.21±0.24	51.83±11.79	65.54±13.83	9.10±1.97^a	60.79±16.01^a
DR组	109	5.64±1.90^ab	4.26±1.24^ab	104.90±26.06^ab	86.67±20.66^ab	13.78±2.18^ab	96.62±25.29^ab
F		371.272^＊＊	502.703^＊＊	278.583^＊＊	62.834^＊＊	546.075^＊＊	151.834^＊＊

组别	n	circRNA-ZNF532	circRNA-HIPK3	VEGF/（ng/L）	IL-1β/（ng/L）	TNF-α/（ng/L）	IL-6/（ng/L）
对照组	65	1.60±0.31	1.15±0.29	48.63±13.36	62.20±12.17	3.77±1.42	49.96±9.80
DM组	110	1.65±0.34	1.21±0.24	51.83±11.79	65.54±13.83	9.10±1.97^a	60.79±16.01^a
DR组	109	5.64±1.90^ab	4.26±1.24^ab	104.90±26.06^ab	86.67±20.66^ab	13.78±2.18^ab	96.62±25.29^ab
F		371.272^＊＊	502.703^＊＊	278.583^＊＊	62.834^＊＊	546.075^＊＊	151.834^＊＊

变量	β	SE	Wald χ²	P	OR（95%CI）
糖尿病病程	0.802	0.263	9.299	<0.001	2.230（1.332~3.734）
高血压	0.502	0.206	5.938	0.010	1.652（1.103~2.474）
HbA1c	0.904	0.277	10.651	<0.001	2.469（1.435~4.250）
circRNA- ZNF532	0.576	0.263	4.797	0.013	1.779（1.062~2.979）
circRNA- HIPK3	0.679	0.251	7.318	<0.001	1.972（1.206~3.225）
VEGF	0.602	0.279	4.656	0.019	1.826（1.057~3.155）
IL-1β	0.346	0.162	4.562	0.023	1.413（1.029~1.942）
FPG	0.264	0.251	1.106	0.365	1.302（0.796~2.130）
TNF-α	0.302	0.268	1.270	0.465	1.353（0.800~2.287）
IL-6	0.291	0.287	1.028	0.598	1.338（0.762~2.348）