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The study of fetal rat model of intra-amniotic isoproterenol injection induced heart dysfunction and phenotypic switch of contractile proteins.

Li Y, Fang J, Hua Y, Wang C, Mu D, Zhou K - Biomed Res Int (2014)

Bottom Line: Intra-amniotic injections of isoproterenol were supplied on E14.5 and E15.5 for fetuses and 7-day continuous intraperitoneal injections were performed for adults.Isoproterenol twice treated fetuses exhibited significant changes in histological evaluation, and mitochondrial damages were significantly severe with increased apoptotic rate.ANP and BNP increased and that of MMP-2 increased in isoproterenol twice treated group compared to control group, without CTGF.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatric Cardiovascular Disease, West China Second University Hospital, Sichuan University, No. 20, 3rd Section, South Renmin Road, Chengdu, Sichuan 610041, China.

ABSTRACT
To establish a reliable isoproterenol induced heart dysfunction fetal rat model and understand the switches of contractile proteins, 45 pregnant rats were divided into 15 mg/kg-once, 15 mg/kg-twice, sham-operated once, sham-operated twice, and control groups. And 18 adult rats were divided into isoproterenol-treated and control groups. H&E staining, Masson staining, and transmission electron microscope were performed. Apoptotic rate assessed by TUNEL analysis and expressions of ANP, BNP, MMP-2, and CTGF of hearts were measured. Intra-amniotic injections of isoproterenol were supplied on E14.5 and E15.5 for fetuses and 7-day continuous intraperitoneal injections were performed for adults. Then echocardiography was performed with M-mode view assessment on E18.5 and 6 weeks later, respectively. Isoproterenol twice treated fetuses exhibited significant changes in histological evaluation, and mitochondrial damages were significantly severe with increased apoptotic rate. ANP and BNP increased and that of MMP-2 increased in isoproterenol twice treated group compared to control group, without CTGF. The isoforms transition of troponin I and myosin heavy chain of fetal heart dysfunction were opposite to adult procedure. The administration of intra-amniotic isoproterenol to fetal rats could induce heart dysfunction and the regulation of contractile proteins of fetuses was different from adult procedure.

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Mitochondrial damages determined with transmission electron microscope in control, sham-operated, and ISO-treated hearts. (a) Presented transmission electron microscope images in control and ISO-treated hearts. Areas of mitochondria were focused. Severe reshaping, severe swelling, and loss of cristae with amorphous dense granules were observed in Iso2 group. 16000x–20000x. (b) Demonstrated FLAMENG index among five experimental groups. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.0001. n = 3/group.
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fig5: Mitochondrial damages determined with transmission electron microscope in control, sham-operated, and ISO-treated hearts. (a) Presented transmission electron microscope images in control and ISO-treated hearts. Areas of mitochondria were focused. Severe reshaping, severe swelling, and loss of cristae with amorphous dense granules were observed in Iso2 group. 16000x–20000x. (b) Demonstrated FLAMENG index among five experimental groups. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.0001. n = 3/group.

Mentions: To further evaluate the impacts of ISO induced cardiac dysfunction on heart microstructure, we examined the ultrastructure of the heart using a transmission electron microscope of fetuses among five groups on E18.5 (Figure 5(a)). The myocardial damages were analyzed based on the myocardial and mitochondria morphological changes (FLAMENG index of Iso1, Iso2, Sham1, Sham2, and Con were 1.28 ± 0.07, 3.05 ± 0.09, 0.61 ± 0.08, 0.91 ± 0.11, and 0.63 ± 0.12, respectively; Figure 5(b)). As shown in Figure 5(a), myocardial vacuolization, degranulation of endoplasmic reticulum, and mitochondrial damages were observed in the fetal hearts of ISO injected groups with a varied extent. Specifically, severe reshaping, severe swelling, and loss of cristae with amorphous dense granules were observed in Iso2 group indicating severe mitochondrial damages. Statistical comparisons revealed that significant differences existed between fetuses with ISO injection and fetuses with sham-operated as well as normal fetuses. However, no difference was recorded on fetuses among sham groups and normal group.


The study of fetal rat model of intra-amniotic isoproterenol injection induced heart dysfunction and phenotypic switch of contractile proteins.

Li Y, Fang J, Hua Y, Wang C, Mu D, Zhou K - Biomed Res Int (2014)

Mitochondrial damages determined with transmission electron microscope in control, sham-operated, and ISO-treated hearts. (a) Presented transmission electron microscope images in control and ISO-treated hearts. Areas of mitochondria were focused. Severe reshaping, severe swelling, and loss of cristae with amorphous dense granules were observed in Iso2 group. 16000x–20000x. (b) Demonstrated FLAMENG index among five experimental groups. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.0001. n = 3/group.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4127273&req=5

fig5: Mitochondrial damages determined with transmission electron microscope in control, sham-operated, and ISO-treated hearts. (a) Presented transmission electron microscope images in control and ISO-treated hearts. Areas of mitochondria were focused. Severe reshaping, severe swelling, and loss of cristae with amorphous dense granules were observed in Iso2 group. 16000x–20000x. (b) Demonstrated FLAMENG index among five experimental groups. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.0001. n = 3/group.
Mentions: To further evaluate the impacts of ISO induced cardiac dysfunction on heart microstructure, we examined the ultrastructure of the heart using a transmission electron microscope of fetuses among five groups on E18.5 (Figure 5(a)). The myocardial damages were analyzed based on the myocardial and mitochondria morphological changes (FLAMENG index of Iso1, Iso2, Sham1, Sham2, and Con were 1.28 ± 0.07, 3.05 ± 0.09, 0.61 ± 0.08, 0.91 ± 0.11, and 0.63 ± 0.12, respectively; Figure 5(b)). As shown in Figure 5(a), myocardial vacuolization, degranulation of endoplasmic reticulum, and mitochondrial damages were observed in the fetal hearts of ISO injected groups with a varied extent. Specifically, severe reshaping, severe swelling, and loss of cristae with amorphous dense granules were observed in Iso2 group indicating severe mitochondrial damages. Statistical comparisons revealed that significant differences existed between fetuses with ISO injection and fetuses with sham-operated as well as normal fetuses. However, no difference was recorded on fetuses among sham groups and normal group.

Bottom Line: Intra-amniotic injections of isoproterenol were supplied on E14.5 and E15.5 for fetuses and 7-day continuous intraperitoneal injections were performed for adults.Isoproterenol twice treated fetuses exhibited significant changes in histological evaluation, and mitochondrial damages were significantly severe with increased apoptotic rate.ANP and BNP increased and that of MMP-2 increased in isoproterenol twice treated group compared to control group, without CTGF.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatric Cardiovascular Disease, West China Second University Hospital, Sichuan University, No. 20, 3rd Section, South Renmin Road, Chengdu, Sichuan 610041, China.

ABSTRACT
To establish a reliable isoproterenol induced heart dysfunction fetal rat model and understand the switches of contractile proteins, 45 pregnant rats were divided into 15 mg/kg-once, 15 mg/kg-twice, sham-operated once, sham-operated twice, and control groups. And 18 adult rats were divided into isoproterenol-treated and control groups. H&E staining, Masson staining, and transmission electron microscope were performed. Apoptotic rate assessed by TUNEL analysis and expressions of ANP, BNP, MMP-2, and CTGF of hearts were measured. Intra-amniotic injections of isoproterenol were supplied on E14.5 and E15.5 for fetuses and 7-day continuous intraperitoneal injections were performed for adults. Then echocardiography was performed with M-mode view assessment on E18.5 and 6 weeks later, respectively. Isoproterenol twice treated fetuses exhibited significant changes in histological evaluation, and mitochondrial damages were significantly severe with increased apoptotic rate. ANP and BNP increased and that of MMP-2 increased in isoproterenol twice treated group compared to control group, without CTGF. The isoforms transition of troponin I and myosin heavy chain of fetal heart dysfunction were opposite to adult procedure. The administration of intra-amniotic isoproterenol to fetal rats could induce heart dysfunction and the regulation of contractile proteins of fetuses was different from adult procedure.

Show MeSH
Related in: MedlinePlus