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. 2015:2015:242517.
doi: 10.1155/2015/242517. Epub 2015 Mar 3.

NaoXinTong Inhibits the Development of Diabetic Retinopathy in db/db Mice

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NaoXinTong Inhibits the Development of Diabetic Retinopathy in db/db Mice

Mengyang Liu et al. Evid Based Complement Alternat Med. 2015.

Abstract

Buchang NaoXinTong capsule (NXT) is a Chinese Materia Medica standardized product extracted from 16 Chinese traditional medical herbs and widely used for treatment of patients with cerebrovascular and cardiovascular diseases in China. Formation of microaneurysms plays an important role in the development of diabetic retinopathy. In this study, we investigated if NXT can protect diabetic mice against the development of diabetic retinopathy. The db/db mice (~6 weeks old), a diabetic animal model, were divided into two groups and fed normal chow or plus NXT for 14 weeks. During the treatment, fasting blood glucose levels were monthly determined. After treatment, retinas were collected to determine retinal thickness, accumulation of carbohydrate macromolecules, and caspase-3 (CAS-3) expression. Our results demonstrate that administration of NXT decreased fasting blood glucose levels. Associated with the decreased glucose levels, NXT blocked the diabetes-induced shrink of multiple layers, such as photoreceptor layer and outer nuclear/plexiform layers, in the retina. NXT also inhibited the diabetes-induced expression of CAS-3 protein and mRNA, MMP-2/9 and TNFα mRNA, accumulation of carbohydrate macromolecules, and formation of acellular capillaries in the retina. Taken together, our study shows that NXT can inhibit the development of diabetic retinopathy and suggests a new potential application of NXT in clinic.

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Figures

Figure 1
Figure 1
NXT inhibits the accumulation of carbohydrate macromolecules and the formation of acellular capillaries in retinal vasculature in db/db mice. (a) At the end of the study, mouse eyes were collected and the retinal vascular network was prepared followed by PAS staining and photograph under a microscope as described in Section 2. The representative images from each group were presented. Black arrows indicate acellular capillaries in the retinal vasculature. Bars: 1 mm and 50 μm in the up and middle panels, respectively. (b) The density of PAS staining was quantified by the Photoshop software. (c) Quantitation of acellular capillaries in the retina. P < 0.05 (n ≥ 3).
Figure 2
Figure 2
NXT corrects the retinal abnormalities in db/db mice. Mouse eyeballs were collected at the end of the study and fixed in 4% PFA/PBS. The 5 μm frozen cross sections were prepared and used to conduct HE staining as described in Section 2. The representative images from each group were presented. Bar: 100 μm.
Figure 3
Figure 3
NXT prevents the reduction of thickness of whole retina and sublayers in retina in db/db mice. After HE staining and photographing, the thickness of whole retina and sublayers in retina was quantified. OPL: the outer plexiform layer; ONL: outer nuclear layer, IS + OS: photoreceptor layer; RPE: retinal pigment epithelium layer. P < 0.05 (n ≥ 5).
Figure 4
Figure 4
NXT inhibits diabetes-induced CAS-3 expression. (a) The frozen sections of mouse eyeballs from each group were prepared and CAS-3 protein expression was determined by immunofluorescent staining as described in Section 2. Bars: 50 μm. (b) CAS-3 mRNA expression in the retinas was determined by real time RT-PCR analysis. (c) TNF-α, MMP-2 and MMP-9 mRNA expression in the retinas was determined by real time RT-PCR analysis. P < 0.05 (n ≥ 5).

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