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A new oxidative stress model, 2,2-azobis(2-amidinopropane) dihydrochloride induces cardiovascular damages in chicken embryo.

He RR, Li Y, Li XD, Yi RN, Wang XY, Tsoi B, Lee KK, Abe K, Yang X, Kurihara H - PLoS ONE (2013)

Bottom Line: At this concentration, AAPH was found to significantly reduce the density of blood vessel plexus that was developed in the chorioallantoic membrane (CAM) of HH 35 chick embryos.These results implied that AAPH-induced oxidative stress could impair the whole developmental processes associated with vasculogenesis and angiogenesis.In conclusion, oxidative stress, induced by AAPH, could lead to damage of the cardiovascular system in the developing chick embryo.

View Article: PubMed Central - PubMed

Affiliation: Pharmacy College, Jinan University, Guangzhou, People's Republic of China.

ABSTRACT
It is now well established that the developing embryo is very sensitive to oxidative stress, which is a contributing factor to pregnancy-related disorders. However, little is known about the effects of reactive oxygen species (ROS) on the embryonic cardiovascular system due to a lack of appropriate ROS control method in the placenta. In this study, a small molecule called 2,2-azobis(2-amidinopropane) dihydrochloride (AAPH), a free radicals generator, was used to study the effects of oxidative stress on the cardiovascular system during chick embryo development. When nine-day-old (stage HH 35) chick embryos were treated with different concentrations of AAPH inside the air chamber, it was established that the LD50 value for AAPH was 10 µmol/egg. At this concentration, AAPH was found to significantly reduce the density of blood vessel plexus that was developed in the chorioallantoic membrane (CAM) of HH 35 chick embryos. Impacts of AAPH on younger embryos were also examined and discovered that it inhibited the development of vascular plexus on yolk sac in HH 18 embryos. AAPH also dramatically repressed the development of blood islands in HH 3+ embryos. These results implied that AAPH-induced oxidative stress could impair the whole developmental processes associated with vasculogenesis and angiogenesis. Furthermore, we observed heart enlargement in the HH 40 embryo following AAPH treatment, where the left ventricle and interventricular septum were found to be thickened in a dose-dependent manner due to myocardiac cell hypertrophy. In conclusion, oxidative stress, induced by AAPH, could lead to damage of the cardiovascular system in the developing chick embryo. The current study also provided a new developmental model, as an alternative for animal and cell models, for testing small molecules and drugs that have anti-oxidative activities.

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Related in: MedlinePlus

Embryo mortality rate in presence of different AAPH concentration.(A) AAPH (0–40 µmol/egg) were injected into the air chamber of eggs containing 9-day-old (stage HH 35) chick embryos and incubated for 24 hours. (B) Stage HH 35 chick embryos were treated with AAPH (10 µmol/egg) and cultured for 2–48 hours. The experiments were performed in triplicates with 20 eggs assigned for each group. Embryos without heart beat were deemed dead.
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pone-0057732-g001: Embryo mortality rate in presence of different AAPH concentration.(A) AAPH (0–40 µmol/egg) were injected into the air chamber of eggs containing 9-day-old (stage HH 35) chick embryos and incubated for 24 hours. (B) Stage HH 35 chick embryos were treated with AAPH (10 µmol/egg) and cultured for 2–48 hours. The experiments were performed in triplicates with 20 eggs assigned for each group. Embryos without heart beat were deemed dead.

Mentions: Serial dilution of water-soluble AAPH (40, 30, 20, 10, 5, 2.5 µmol/egg) were injected into the air chamber of 9-day-old (HH 35) chick embryos and incubated for 24 h. In the control, the eggs were injected with simple saline. Embryos were deemed dead if they did not exhibit heartbeat. The mortality rate of AAPH treated embryos appeared to increase in a dose dependent manner, as shown in Figure 1A. The LD50 of AAPH treatment was calculated as 10 µmol/egg. The mortality rate of embryos exposed to LD50 of AAPH for 2, 4, 8, 12, 24, 36 and 48 hours was presented in Figure 1B. It was found that the mortality rate rose dramatically when AAPH treatment time was extended from 12 hours onwards.


A new oxidative stress model, 2,2-azobis(2-amidinopropane) dihydrochloride induces cardiovascular damages in chicken embryo.

He RR, Li Y, Li XD, Yi RN, Wang XY, Tsoi B, Lee KK, Abe K, Yang X, Kurihara H - PLoS ONE (2013)

Embryo mortality rate in presence of different AAPH concentration.(A) AAPH (0–40 µmol/egg) were injected into the air chamber of eggs containing 9-day-old (stage HH 35) chick embryos and incubated for 24 hours. (B) Stage HH 35 chick embryos were treated with AAPH (10 µmol/egg) and cultured for 2–48 hours. The experiments were performed in triplicates with 20 eggs assigned for each group. Embryos without heart beat were deemed dead.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585800&req=5

pone-0057732-g001: Embryo mortality rate in presence of different AAPH concentration.(A) AAPH (0–40 µmol/egg) were injected into the air chamber of eggs containing 9-day-old (stage HH 35) chick embryos and incubated for 24 hours. (B) Stage HH 35 chick embryos were treated with AAPH (10 µmol/egg) and cultured for 2–48 hours. The experiments were performed in triplicates with 20 eggs assigned for each group. Embryos without heart beat were deemed dead.
Mentions: Serial dilution of water-soluble AAPH (40, 30, 20, 10, 5, 2.5 µmol/egg) were injected into the air chamber of 9-day-old (HH 35) chick embryos and incubated for 24 h. In the control, the eggs were injected with simple saline. Embryos were deemed dead if they did not exhibit heartbeat. The mortality rate of AAPH treated embryos appeared to increase in a dose dependent manner, as shown in Figure 1A. The LD50 of AAPH treatment was calculated as 10 µmol/egg. The mortality rate of embryos exposed to LD50 of AAPH for 2, 4, 8, 12, 24, 36 and 48 hours was presented in Figure 1B. It was found that the mortality rate rose dramatically when AAPH treatment time was extended from 12 hours onwards.

Bottom Line: At this concentration, AAPH was found to significantly reduce the density of blood vessel plexus that was developed in the chorioallantoic membrane (CAM) of HH 35 chick embryos.These results implied that AAPH-induced oxidative stress could impair the whole developmental processes associated with vasculogenesis and angiogenesis.In conclusion, oxidative stress, induced by AAPH, could lead to damage of the cardiovascular system in the developing chick embryo.

View Article: PubMed Central - PubMed

Affiliation: Pharmacy College, Jinan University, Guangzhou, People's Republic of China.

ABSTRACT
It is now well established that the developing embryo is very sensitive to oxidative stress, which is a contributing factor to pregnancy-related disorders. However, little is known about the effects of reactive oxygen species (ROS) on the embryonic cardiovascular system due to a lack of appropriate ROS control method in the placenta. In this study, a small molecule called 2,2-azobis(2-amidinopropane) dihydrochloride (AAPH), a free radicals generator, was used to study the effects of oxidative stress on the cardiovascular system during chick embryo development. When nine-day-old (stage HH 35) chick embryos were treated with different concentrations of AAPH inside the air chamber, it was established that the LD50 value for AAPH was 10 µmol/egg. At this concentration, AAPH was found to significantly reduce the density of blood vessel plexus that was developed in the chorioallantoic membrane (CAM) of HH 35 chick embryos. Impacts of AAPH on younger embryos were also examined and discovered that it inhibited the development of vascular plexus on yolk sac in HH 18 embryos. AAPH also dramatically repressed the development of blood islands in HH 3+ embryos. These results implied that AAPH-induced oxidative stress could impair the whole developmental processes associated with vasculogenesis and angiogenesis. Furthermore, we observed heart enlargement in the HH 40 embryo following AAPH treatment, where the left ventricle and interventricular septum were found to be thickened in a dose-dependent manner due to myocardiac cell hypertrophy. In conclusion, oxidative stress, induced by AAPH, could lead to damage of the cardiovascular system in the developing chick embryo. The current study also provided a new developmental model, as an alternative for animal and cell models, for testing small molecules and drugs that have anti-oxidative activities.

Show MeSH
Related in: MedlinePlus