Wedelolctone upregulates miR-190 expression to inhibit human retinal vascular endothelial cell apoptosis induced by high glucose

doi:10.3877/cma.j.issn.2095-1221.2023.06.003

Abstract

Abstract:

Objective

To investigate the effects and molecular mechanisms of wedelolactone (WEL) on injury of human retinal vascular endothelial cells (HRECs) induced by high glucose (HG) .

Methods

After being treated with low (10 μmol/L), medium (20 μmol/ L) and high (40 μmol/ L) doses of WEL (WEL-L, WEL-M and WEL-H) or transfected with miR- 190 mimics/ inhibitors, the cell injury model was established by incubating HRECs with 30 mmol/L glucose for 24 h. The kit was used to detect cellular reactive oxygen species (ROS), superoxide dismutase (SOD) and malondialdehyde (MDA) content; cell apoptosis was analyzed by flow cytometry; miR-190 expression was detected by RT-qPCR. Independent sample t test and one-way analysis of variance were used for statistical comparison.

Results

Low, medium and high doses of WEL could reduce HG induced apoptosis rate of HRECs [ (24.22 ± 2.16) %, (17.17 ± 1.19) %, (9.89 ± 0.86) % vs (31.67 ± 3.02) %], MDA content and ROS level, and increased the expression of miR-190 (0.28 ± 0.03 vs 0.42 ± 0.04, 0.59 ± 0.04, 0.72 ± 0.06) and SOD activity (P < 0.01). Overexpression of miR- 190 decreased apoptosis rate [ (32.28 ± 3.03) % vs (12.98 ± 1.06) %], MDA content and ROS level of HRECs induced by HG, and increased SOD activity (P < 0.01). Down-regulation of miR-190 reversed the effects of WEL on apoptosis rate [ (9.79 ± 0.87) % vs (25.57 ± 2.49) %], MDA content, ROS level and SOD activity of HRECs induced by HG (P < 0.01) .

Conclusion

Wedelolactone alleviates oxidative stress injury and apoptosis of HRECs induced by HG, and the mechanism is related to up-regulation miR-190.

Key words: Wedelolactone, Retinal vascular endothelial cells, Apoptosis, Oxidative stress, MiR-190

Lingling Yu, Ni Peng, Xiaohu Liu, Conghui Liu. Wedelolctone upregulates miR-190 expression to inhibit human retinal vascular endothelial cell apoptosis induced by high glucose[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2023, 13(06): 339-345.

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URL: https://zhxbygxbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-1221.2023.06.003

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Figures/Tables 11

References 24

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基因	序列（5'→3'）	预期扩增产物长度（bp）
U6（内参）	上游GCGCGTCGTGAAGCGTTC	98
	下游GTGCAGGGTCCGAGGT
miR-190	上游GGTCTTTGATGATGATTCTGG	75
	下游CTAGGCACAGTATTGAAGGTT

基因	序列（5'→3'）	预期扩增产物长度（bp）
U6（内参）	上游GCGCGTCGTGAAGCGTTC	98
	下游GTGCAGGGTCCGAGGT
miR-190	上游GGTCTTTGATGATGATTCTGG	75
	下游CTAGGCACAGTATTGAAGGTT

分组	实验次数	ROS水平（%）	SOD活性（U/mg）	MDA含量（nmol/mg）
对照	3	100.00 ± 0.00	127.70 ± 11.52	3.15 ± 0.37
HG	3	340.74 ± 20.77^a	34.86 ± 3.52^a	19.96 ± 1.58^a
HG+WEL-L	3	265.01 ± 21.56^b	61.52 ± 5.08^b	14.98 ± 1.12^b
HG+WEL-M	3	203.85 ± 16.55^bc	81.62 ± 6.36^bc	10.15 ± 0.87^bc
HG+WEL-H	3	136.91 ± 14.25^bcd	101.79 ± 10.33^bcd	5.21 ± 0.48^bcd
F值		308.830	181.117	441.948
P值		< 0.001	< 0.001	< 0.001

分组	实验次数	ROS水平（%）	SOD活性（U/mg）	MDA含量（nmol/mg）
对照	3	100.00 ± 0.00	127.70 ± 11.52	3.15 ± 0.37
HG	3	340.74 ± 20.77^a	34.86 ± 3.52^a	19.96 ± 1.58^a
HG+WEL-L	3	265.01 ± 21.56^b	61.52 ± 5.08^b	14.98 ± 1.12^b
HG+WEL-M	3	203.85 ± 16.55^bc	81.62 ± 6.36^bc	10.15 ± 0.87^bc
HG+WEL-H	3	136.91 ± 14.25^bcd	101.79 ± 10.33^bcd	5.21 ± 0.48^bcd
F值		308.830	181.117	441.948
P值		< 0.001	< 0.001	< 0.001

分组	实验次数	凋亡率（%）	cleaved caspase-3	cleaved caspase-9
Con	3	6.83 ± 0.59	0.22 ± 0.02	0.12 ± 0.02
HG	3	31.67 ± 3.02^a	0.71 ± 0.06^a	0.58 ± 0.04^a
HG+WEL-L	3	24.22 ± 2.16^b	0.57 ± 0.04^b	0.41 ± 0.03^b
HG+WEL-M	3	17.17 ± 1.19^bc	0.43 ± 0.04^bc	0.29 ± 0.03^bc
HG+WEL-H	3	9.89 ± 0.86^bcd	0.31 ± 0.03^bcd	0.17 ± 0.02^bcd
F值		287.833	215.111	372.107
P值		< 0.01	< 0.01	< 0.01

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