[The mediation of HIF-1α siRNA to the leukocyte adhesion and myeloid cell's activity in rat's retina under early stage of diabetic retinopathy].

Abstract

OBJECTIVE:

To investigate the mediation of HIF-1α siRNA to the leukocyte adhesion and myeloid cell's activity in rat's retina at early stage of diabetic retinopathy.

METHODS:

Experimental study. HIF-1α specific siRNA expression vector pSUPERH1-siHIF1α was constructed by gene recombination. The rat diabetic model was induced by intraperitoneal injection of streptozotocin. After 2 months of diabetes induction, 27 diabetic rats were randomly chosen and assigned to 3 groups, including diabetes and phosphate buffered saline (PBS) vitreous injection group (group B), diabetes and pSUPERH1-siHIF1α vitreous transfect group (group C) and diabetes and pSUPER-retro vitreous transfect group (group D). Each group had 9 rats. Nine age matched health rats were chosen as control group (group A). Retinal leukostasis was quantified with acridine orange leukocyte fluorography. Retinal myeloid cell activity was measured by enzyme linked immunosorbent assay of myeloperoxidase (MPO). The differences of the mean values among the four groups were analyzed by one-factor analysis of variance. The multiple comparisons of the mean values among the four groups were analyzed by LSD-t analysis.

RESULTS:

According to the results of the acridine orange leukocyte fluorography, the numbers of leukocyte adhesion in the four groups were (47.00 ± 3.60), (155.33 ± 9.01), (76.00 ± 9.05), (142.66 ± 10.26), respectively. The differences among them were significant (F = 116.25, P = 0.00). The number of leukocyte adhesion in the group C was significantly lower than that in group B (LSD-t test, P = 0.00, 95% CI: 3.56-95.10). The levels of retinal MPO in the four groups were (17.24 ± 1.13), (31.32 ± 2.53), (21.35 ± 1.06), (31.33 ± 1.26) µg/L, respectively. The differences among them were significant (F = 58.68, P = 0.00). The level of retinal MPO in the group C was significantly lower than that in group B (LSD-t test, P = 0.00, 95% CI: 6.92-13.01).

CONCLUSIONS:

HIF-1α siRNA may play a role in the mediation of the leukocyte adhesion and myeloid cell's activity in rat's retina at early stage of diabetic retinopathy in vivo.

• Authors: Huping Song, Qi Zhu, Qiong Wu, Saile Song, Hongfen Sun, Xiaoliang Zhou, Hui Liu
• Journal: [Zhonghua yan ke za zhi] Chinese journal of ophthalmology (Zhonghua Yan Ke Za Zhi) Vol. 51 Issue 5 Pg. 351-5 (May 2015) ISSN: 0412-4081 [Print] China
• PMID: 26311695 (Publication Type: Journal Article)

Chemical References

• Fluorescent Dyes
• Hypoxia-Inducible Factor 1, alpha Subunit
• RNA, Small Interfering
• Peroxidase
• Acridine Orange

Topics

• Acridine Orange
• Animals
• Cell Adhesion
• Cell Movement
• Diabetes Mellitus, Experimental (genetics, pathology)
• Diabetic Retinopathy (genetics, pathology)
• Fluorescent Dyes
• Hypoxia-Inducible Factor 1, alpha Subunit (genetics)
• Leukocytes (enzymology, physiology)
• Myeloid Cells (enzymology, physiology)
• Peroxidase (analysis)
• RNA, Small Interfering (physiology)
• Rats
• Retina (enzymology, pathology)