Protective effect of Qideng Mingmu capsule on retinal vessels in mice with oxygen-induced retinopathy

Authors: Liu Chunmeng,  Ding Shan,  Dong Xuewen,  Zhao Dandan,  Pu Siyuan,  Pei Li,  Zhang Fuwen
DOI: 10.3760/cma.j.cn115989-20220426-00181
Published 2024-05-10
Cite as Chin J Exp Ophthalmol, 2024, 42(5): 428-435.

Abstract                              [Download PDF] [Read Full Text]

Objective

To investigate the effect of Qideng Mingmu capsule on the formation and remodeling of retinal neovascularization in mice with oxygen-induced retinopathy (OIR).

Methods

Thirty-six postnatal day 7 (P7)SPF grade C57BL/6J pups were divided into normal group, OIR group, Qideng Mingmu capsule group and apatinib group by random number table method, with 9 mice in each group.The mice in the normal group were raised in normal environment.The mice in the other three groups were fed in hyperoxic environment of (75±2)% oxygen concentration for 5 days from P7 to P12 and then were fed in normal environment for 5 days from P12 to P17 to establish the OIR model.From P12, mice in Qideng Mingmu capsule group and apatinib group were given intragastric administration of Qideng Mingmu capsule (900 mg/kg) and vascular endothelial growth factor receptor 2 inhibitor apatinib (70 mg/kg) respectively, once a day for 5 consecutive days.On P17, paraffin sections of mouse eyeballs were made and stained with hematoxylin-eosin to count the number of vascular endothelial cells that broke through the internal limiting membrane.The retinal slices were prepared and stained with FITC-dextran to quantify the retinal non-perfusion area, neovascularization density and total vascular density.The distribution and fluorescence intensity of retinal vascular endothelial cell marker CD31 and pericyte marker α-smooth muscle actin (α-SMA) were observed by double immunofluorescence staining.Immunohistochemical staining was used to detect the expression and distribution of retinal hypoxia inducible factor-1α (HIF-1α) and vascular endothelial cadherin (VE-cadherin).The use and care of animals were in accordance with the Regulations on the Management of Laboratory Animals issued by the Ministry of Science and Technology.This study was approved by the Animal Ethics Committee of Chengdu University of Traditional Chinese Medicine (No.2019-30).

Results

The number of vascular endothelial cells breaking through the internal limiting membrane in normal group, OIR group, Qideng Mingmu capsule group and apatinib group were (2.83±4.40), (37.33±5.43), (23.83±6.79) and (14.00±9.34), respectively, with a statistically significant overall difference (F=28.313, P<0.001).There were more vascular endothelial cells breaking through internal limiting membrane in OIR group than in normal group, Qideng Mingmu capsule group and apatinib group, showing statistically significant differences (all at P<0.05).In the observation of mouse retinal slices, there were large non-perfusion areas, neovascularization buds and disordered distribution of blood vessels in OIR group.The distribution of blood vessels was more uniform and the areas of non-perfusion and neovascularization were smaller in Qideng Mingmu capsule group and apatinib group than in OIR group.The relative area of central retinal non-perfusion area and neovascularization density were significantly lower in normal group, Qideng Mingmu capsule group and apatinib group than in OIR group (all at P<0.05).The immunofluorescence intensity of CD31 and the absorbance value of HIF-1α were significantly lower, and the immunofluorescence intensity of α-SMA and the absorbance value of VE-cadherin were significantly higher in normal group, Qideng Mingmu capsule group and apatinib group than in OIR group (all at P<0.05).

Conclusions

Qideng Mingmu capsule can inhibit retinal neovascularization formation, increase vascular pericyte coverage, relieve retinal hypoxia and increase vascular integrity in OIR mice.It can protect the retinal vessels of OIR mice.

Key words:

Retinal neovascularization; Oxygen-induced retinopathy; Mouse; Qideng Mingmu capsule; Apatinib

Contributor Information

Liu Chunmeng

Eye School of Chengdu University of Traditional Chinese Medicine, Key Laboratory of Sichuan Province Ophthalmopathy Prevention &

Cure and Visual Function Protection, Chengdu 610075, China

Ding Shan

Eye School of Chengdu University of Traditional Chinese Medicine, Key Laboratory of Sichuan Province Ophthalmopathy Prevention &

Cure and Visual Function Protection, Chengdu 610075, China

Dong Xuewen

Eye School of Chengdu University of Traditional Chinese Medicine, Key Laboratory of Sichuan Province Ophthalmopathy Prevention &

Cure and Visual Function Protection, Chengdu 610075, China

Zhao Dandan

Eye School of Chengdu University of Traditional Chinese Medicine, Key Laboratory of Sichuan Province Ophthalmopathy Prevention &

Cure and Visual Function Protection, Chengdu 610075, China

Pu Siyuan

Eye School of Chengdu University of Traditional Chinese Medicine, Key Laboratory of Sichuan Province Ophthalmopathy Prevention &

Cure and Visual Function Protection, Chengdu 610075, China

Pei Li

Eye School of Chengdu University of Traditional Chinese Medicine, Key Laboratory of Sichuan Province Ophthalmopathy Prevention &

Cure and Visual Function Protection, Chengdu 610075, China

Zhang Fuwen

Eye School of Chengdu University of Traditional Chinese Medicine, Key Laboratory of Sichuan Province Ophthalmopathy Prevention &

Cure and Visual Function Protection, Chengdu 610075, China

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