Limits...
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.

Show MeSH
Effects of mini-αA on CHOP and XBP1 expression level. (A) Western blot analysis was performed to detect the expression level of CHOP protein in each group; (B) RT-qPCR was used to detect the expression level of CHOP mRNA in each group. CTL: normal ARPE-19 cells; * p < 0.05; (C) Western blot was performed to detect XBP1 protein expression level in each group; (D) RT-qPCR was used to detect the expression level of XBP1 mRNA in each group. NaIO3 induction elevated the expression level of XBP1 in RPE cells, while mini-αA treatment inhibited the increase caused by NaIO3 induction. CTL: normal ARPE-19 cells, NaIO3: cells induced with NaIO3; NaIO3 + mini-αA: cells treated first with mini-αA and then with NaIO3. GAPDH was used as the internal. * p < 0.05.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4307325&req=5

ijms-16-01644-f005: Effects of mini-αA on CHOP and XBP1 expression level. (A) Western blot analysis was performed to detect the expression level of CHOP protein in each group; (B) RT-qPCR was used to detect the expression level of CHOP mRNA in each group. CTL: normal ARPE-19 cells; * p < 0.05; (C) Western blot was performed to detect XBP1 protein expression level in each group; (D) RT-qPCR was used to detect the expression level of XBP1 mRNA in each group. NaIO3 induction elevated the expression level of XBP1 in RPE cells, while mini-αA treatment inhibited the increase caused by NaIO3 induction. CTL: normal ARPE-19 cells, NaIO3: cells induced with NaIO3; NaIO3 + mini-αA: cells treated first with mini-αA and then with NaIO3. GAPDH was used as the internal. * p < 0.05.

Mentions: To examine the relationship between unfolded protein response (UPR) and mini-αA’s protective function against NaIO3-induced RPE cell apoptosis, Western blot analysis was performed to detect CHOP expression levels in cells induced by NaIO3 at concentrations of 2, 2.5, 3 and 3.5 mM. We discovered that, compared with normal untreated RPE cells, there was a marked increase in CHOP protein expression in cells treated with NaIO3, and the expression level rose as the concentration of NaIO3 rose. After being treated with mini-αA at 10, 15 or 20 μM, CHOP protein expression levels of RPE cells induced by 3.5 mM NaIO3 were evidently repressed, with 20 μM mini-αA showing the best inhibitive effect (Figure 5A). In addition, RT-qPCR was used to detect expression levels of CHOP mRNA in each group. The result was the same as that of the Western blot analysis. The CHOP mRNA expression levels in cells induced by NaIO3 at 2, 2.5, 3 or 3.5 mM were much higher than that of normal cells, and the mRNA expression level rose as the concentration of NaIO3 rose. After mini-αA treatment, the mRNA expression level decreased, with 20 μM mini-αA showing the greatest inhibition (Figure 5B).


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)

Effects of mini-αA on CHOP and XBP1 expression level. (A) Western blot analysis was performed to detect the expression level of CHOP protein in each group; (B) RT-qPCR was used to detect the expression level of CHOP mRNA in each group. CTL: normal ARPE-19 cells; * p < 0.05; (C) Western blot was performed to detect XBP1 protein expression level in each group; (D) RT-qPCR was used to detect the expression level of XBP1 mRNA in each group. NaIO3 induction elevated the expression level of XBP1 in RPE cells, while mini-αA treatment inhibited the increase caused by NaIO3 induction. CTL: normal ARPE-19 cells, NaIO3: cells induced with NaIO3; NaIO3 + mini-αA: cells treated first with mini-αA and then with NaIO3. GAPDH was used as the internal. * p < 0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4307325&req=5

ijms-16-01644-f005: Effects of mini-αA on CHOP and XBP1 expression level. (A) Western blot analysis was performed to detect the expression level of CHOP protein in each group; (B) RT-qPCR was used to detect the expression level of CHOP mRNA in each group. CTL: normal ARPE-19 cells; * p < 0.05; (C) Western blot was performed to detect XBP1 protein expression level in each group; (D) RT-qPCR was used to detect the expression level of XBP1 mRNA in each group. NaIO3 induction elevated the expression level of XBP1 in RPE cells, while mini-αA treatment inhibited the increase caused by NaIO3 induction. CTL: normal ARPE-19 cells, NaIO3: cells induced with NaIO3; NaIO3 + mini-αA: cells treated first with mini-αA and then with NaIO3. GAPDH was used as the internal. * p < 0.05.
Mentions: To examine the relationship between unfolded protein response (UPR) and mini-αA’s protective function against NaIO3-induced RPE cell apoptosis, Western blot analysis was performed to detect CHOP expression levels in cells induced by NaIO3 at concentrations of 2, 2.5, 3 and 3.5 mM. We discovered that, compared with normal untreated RPE cells, there was a marked increase in CHOP protein expression in cells treated with NaIO3, and the expression level rose as the concentration of NaIO3 rose. After being treated with mini-αA at 10, 15 or 20 μM, CHOP protein expression levels of RPE cells induced by 3.5 mM NaIO3 were evidently repressed, with 20 μM mini-αA showing the best inhibitive effect (Figure 5A). In addition, RT-qPCR was used to detect expression levels of CHOP mRNA in each group. The result was the same as that of the Western blot analysis. The CHOP mRNA expression levels in cells induced by NaIO3 at 2, 2.5, 3 or 3.5 mM were much higher than that of normal cells, and the mRNA expression level rose as the concentration of NaIO3 rose. After mini-αA treatment, the mRNA expression level decreased, with 20 μM mini-αA showing the greatest inhibition (Figure 5B).

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