The effect of miR-3202 on human retinal vascular endothelial cell injury induced by high glucose

doi:10.3877/cma.j.issn.2095-1221.2021.03.004

Abstract

Abstract:

Objective

To investigate the effect of microRNA-3202 (miR-3202) on apoptosis induced by high glucose in human retinal vascular endothelial cells (HRCEC) and its molecular mechanisms.

Methods

HRCEC was cultured in vitro and divided into eight groups, including low glucose group, high glucose group, high glucose+miR-NC group, high glucose+miR-3202 group, high glucose+si-NC group, high glucose+si-ILK group, high sugar + miR -3202+pcDNA3.1 group, high glucose+miR-3202+pcDNA3.1-ILK group. The expression of miR-3202 and ILK in HRCEC was determined by real-time quantitative PCR (qRT-PCR) and Western blot, and the apoptotic rate was determined by flow cytometry. The dual luciferase reporter gene assay and Western blot assay were used to verified the relationship between miR-3202 and ILK. Also, The expression of Bax and Bcl-2 protein in HRCEC was determined by Western blot.

Results

Compared with the low glucose group, the expression level of miR-3202 in the HRCEC of the high glucose group was significantly decreased (0.95±0.09 vs 0.21±0.02, P < 0.05), and the expression level of ILK (0.30±0.03 vs 0.86±0.08) and the rate of cell apoptosis were significantly increased (8.54%±1.03% vs 28.53%± 3.25%, P < 0.05). Compared with the high glucose + miR-NC group, the apoptotic rate of the high glucose + miR-3202 group (28.86%±2.45% vs 12.67%±1.15%) and the expression level of Bax were significantly decreased (1.12±0.11 vs 0.36±0.03, P < 0.05), however,the expression level of Bcl-2 significantly increased (0.23±0.02 vs 0.77±0.07, P < 0.05). Compared with the high glucose + si-NC group, the apoptotic rate of the high glucose + si-ILK group (28.86%±2.45% vs 13.46%±1.24%, P < 0.05) and the expression level of Bax were both significantly decreased (1.10±0.11 vs 0.47±0.05, P < 0.05), while the expression level of Bcl-2 was significantly increased (0.23±0.02 vs 0.64±0.06, P < 0.05). MiR-3202 may negatively regulate the expression of the target gene ILK. Overexpression of ILK reversed the effect of miR-3202 on high glucose-induced apoptosis of HRCEC.

Conclusion

high glucose can induce the apoptosis of HRCEC and miR-3202 may inhibit it by down-regulating the expression of ILK.

Key words: miR-3202, Integrin-linked kinase, Diabetic retinopathy, Apoptosis

Hong Luo, Changliang Yuan, Lan Chen. The effect of miR-3202 on human retinal vascular endothelial cell injury induced by high glucose[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2021, 11(03): 155-160.

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Figures/Tables 13

References 18

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基因	序列（5'→3'）	预期扩增产物长度（bp）
miR-3202	上游ATGGAAGGGAGAAGAGC	120
	下游GCTCGTCTCTGATCTCGCG
	RT TGGAAGGGAGAAGAGCTTTATATAAAG
U6	上游ATTGGAACGATACAGAGAAGATT	96
U6	下游GGAACGCTTCACGAATTTG	96

基因	序列（5'→3'）	预期扩增产物长度（bp）
miR-3202	上游ATGGAAGGGAGAAGAGC	120
	下游GCTCGTCTCTGATCTCGCG
	RT TGGAAGGGAGAAGAGCTTTATATAAAG
U6	上游ATTGGAACGATACAGAGAAGATT	96
U6	下游GGAACGCTTCACGAATTTG	96

分组	实验次数	miR-3202	ILK蛋白
低糖	6	0.95±0.09	0.30±0.03
高糖	6	0.21±0.02	0.86±0.08
t值		19.661	16.055
P值		<0.001	<0.001

分组	实验次数	miR-3202	ILK蛋白
低糖	6	0.95±0.09	0.30±0.03
高糖	6	0.21±0.02	0.86±0.08
t值		19.661	16.055
P值		<0.001	<0.001

分组	实验次数	miR-3202	Bax蛋白	Bcl-2蛋白	凋亡率（%）
高糖+miR-NC	6	0.20±0.02	1.12±0.11	0.23±0.02	28.86±2.45
高糖+miR-3202	6	0.83±0.08	0.36±0.03	0.77±0.07	12.67±1.15
t值		18.714	16.327	18.169	14.653
P值		<0.001	<0.001	<0.001	<0.001

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