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Protection of retina by mini-αA in NaIO3-induced retinal pigment epithelium degeneration mice.

Zhang J, Zhao X, Cai Y, Li Y, Yu X, Lu L - Int J Mol Sci (2015)

Bottom Line: Further investigation into how mini-αA provided protection against NaIO3-induced RPE cell apoptosis showed that mini-αA reduced NaIO3-induced RPE cell apoptosis and autophagy.In addition, unfolded protein response was also involved in the protective effects of mini-αA against NaIO3-induced RPE cell apoptosis. mini-αA can antagonize RPE cell apoptosis induced by NaIO3.A possible mechanism is by inhibition of apoptosis by repressing autophagy and endoplasmic reticulum stress.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China. zhjinglin@126.com.

ABSTRACT

Background: Studies have shown that mini-αA can protect retinal pigment epithelium (RPE) cells from apoptosis. However, no in vivo study concerning the anti-apoptotic function of mini-αA has been conducted yet.

Methods: MTT assay, HE staining and TUNEL assay were used to assess levels of cells, and an animal model was established to examine the protective effects of mini-αA against NaIO3-induced RPE cell apoptosis. Western blot analysis and RT-qPCR were performed to explore the possible mechanism of mini-αA's protective function against NaIO3-induced RPE cell apoptosis.

Results: RESULTS from in vivo and animal experiments showed that mini-αA antagonized NaIO3-induced RPE cell apoptosis. Further investigation into how mini-αA provided protection against NaIO3-induced RPE cell apoptosis showed that mini-αA reduced NaIO3-induced RPE cell apoptosis and autophagy. In addition, unfolded protein response was also involved in the protective effects of mini-αA against NaIO3-induced RPE cell apoptosis.

Conclusions: mini-αA can antagonize RPE cell apoptosis induced by NaIO3. A possible mechanism is by inhibition of apoptosis by repressing autophagy and endoplasmic reticulum stress.

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Related in: MedlinePlus

TUNEL assay was performed 3 days after the establishment of the mouse model of NaIO3-induced retinal degeneration. (A) Almost no TUNEL-positive cells were detected in normal control group; a large number of TUNEL-positive cells were found in the external nuclear layer in the retinas of NaIO3 group (white arrow); several TUNEL-positive cells were observed in the external nuclear layer in mini-αA group (white arrow), but much less than those observed in NaIO3 group. Magnification: 400×; (B) Quantification of dead cells by TUNEL assay from at least five independent experiments. TUNEL-positive cells were counted and data were expressed as percent of total death cells; (C) Western blot analysis of caspase expression levels in each subgroup; (D) Quantification of band intensity of caspase from three independent experiments on (C). * p < 0.05 vs. Control; # p <0.05 vs. NaIO3.
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ijms-16-01644-f003: TUNEL assay was performed 3 days after the establishment of the mouse model of NaIO3-induced retinal degeneration. (A) Almost no TUNEL-positive cells were detected in normal control group; a large number of TUNEL-positive cells were found in the external nuclear layer in the retinas of NaIO3 group (white arrow); several TUNEL-positive cells were observed in the external nuclear layer in mini-αA group (white arrow), but much less than those observed in NaIO3 group. Magnification: 400×; (B) Quantification of dead cells by TUNEL assay from at least five independent experiments. TUNEL-positive cells were counted and data were expressed as percent of total death cells; (C) Western blot analysis of caspase expression levels in each subgroup; (D) Quantification of band intensity of caspase from three independent experiments on (C). * p < 0.05 vs. Control; # p <0.05 vs. NaIO3.

Mentions: Previous studies suggest that the reason why NaIO3 can cause thinning of the retina is that RPE cell apoptosis leads to photoreceptor apoptosis. Three days after successful establishment of the model, a TUNEL assay was performed to assess cell apoptosis in a control group, NaIO3 group and mini-αA group. Results showed that, although there was no significant change in retinal thickness and cellular stratification in each group, only a few TUNEL-positive cells were observed in the retinas of the control group, whereas a large number of TUNEL-positive cells were found in both the external and internal nuclear layers in the retinas of the NaIO3 group. TUNEL-positive cells detected in both the external and internal nuclear layers in the retina of mini-αA group were less than those detected in NaIO3 group, but more than those found in the control group. This explains why the retinas of mini-αA group were thicker than those of NaIO3 group but thinner than those of normal control group in Week two (Figure 3).


Protection of retina by mini-αA in NaIO3-induced retinal pigment epithelium degeneration mice.

Zhang J, Zhao X, Cai Y, Li Y, Yu X, Lu L - Int J Mol Sci (2015)

TUNEL assay was performed 3 days after the establishment of the mouse model of NaIO3-induced retinal degeneration. (A) Almost no TUNEL-positive cells were detected in normal control group; a large number of TUNEL-positive cells were found in the external nuclear layer in the retinas of NaIO3 group (white arrow); several TUNEL-positive cells were observed in the external nuclear layer in mini-αA group (white arrow), but much less than those observed in NaIO3 group. Magnification: 400×; (B) Quantification of dead cells by TUNEL assay from at least five independent experiments. TUNEL-positive cells were counted and data were expressed as percent of total death cells; (C) Western blot analysis of caspase expression levels in each subgroup; (D) Quantification of band intensity of caspase from three independent experiments on (C). * p < 0.05 vs. Control; # p <0.05 vs. NaIO3.
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ijms-16-01644-f003: TUNEL assay was performed 3 days after the establishment of the mouse model of NaIO3-induced retinal degeneration. (A) Almost no TUNEL-positive cells were detected in normal control group; a large number of TUNEL-positive cells were found in the external nuclear layer in the retinas of NaIO3 group (white arrow); several TUNEL-positive cells were observed in the external nuclear layer in mini-αA group (white arrow), but much less than those observed in NaIO3 group. Magnification: 400×; (B) Quantification of dead cells by TUNEL assay from at least five independent experiments. TUNEL-positive cells were counted and data were expressed as percent of total death cells; (C) Western blot analysis of caspase expression levels in each subgroup; (D) Quantification of band intensity of caspase from three independent experiments on (C). * p < 0.05 vs. Control; # p <0.05 vs. NaIO3.
Mentions: Previous studies suggest that the reason why NaIO3 can cause thinning of the retina is that RPE cell apoptosis leads to photoreceptor apoptosis. Three days after successful establishment of the model, a TUNEL assay was performed to assess cell apoptosis in a control group, NaIO3 group and mini-αA group. Results showed that, although there was no significant change in retinal thickness and cellular stratification in each group, only a few TUNEL-positive cells were observed in the retinas of the control group, whereas a large number of TUNEL-positive cells were found in both the external and internal nuclear layers in the retinas of the NaIO3 group. TUNEL-positive cells detected in both the external and internal nuclear layers in the retina of mini-αA group were less than those detected in NaIO3 group, but more than those found in the control group. This explains why the retinas of mini-αA group were thicker than those of NaIO3 group but thinner than those of normal control group in Week two (Figure 3).

Bottom Line: Further investigation into how mini-αA provided protection against NaIO3-induced RPE cell apoptosis showed that mini-αA reduced NaIO3-induced RPE cell apoptosis and autophagy.In addition, unfolded protein response was also involved in the protective effects of mini-αA against NaIO3-induced RPE cell apoptosis. mini-αA can antagonize RPE cell apoptosis induced by NaIO3.A possible mechanism is by inhibition of apoptosis by repressing autophagy and endoplasmic reticulum stress.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China. zhjinglin@126.com.

ABSTRACT

Background: Studies have shown that mini-αA can protect retinal pigment epithelium (RPE) cells from apoptosis. However, no in vivo study concerning the anti-apoptotic function of mini-αA has been conducted yet.

Methods: MTT assay, HE staining and TUNEL assay were used to assess levels of cells, and an animal model was established to examine the protective effects of mini-αA against NaIO3-induced RPE cell apoptosis. Western blot analysis and RT-qPCR were performed to explore the possible mechanism of mini-αA's protective function against NaIO3-induced RPE cell apoptosis.

Results: RESULTS from in vivo and animal experiments showed that mini-αA antagonized NaIO3-induced RPE cell apoptosis. Further investigation into how mini-αA provided protection against NaIO3-induced RPE cell apoptosis showed that mini-αA reduced NaIO3-induced RPE cell apoptosis and autophagy. In addition, unfolded protein response was also involved in the protective effects of mini-αA against NaIO3-induced RPE cell apoptosis.

Conclusions: mini-αA can antagonize RPE cell apoptosis induced by NaIO3. A possible mechanism is by inhibition of apoptosis by repressing autophagy and endoplasmic reticulum stress.

Show MeSH
Related in: MedlinePlus