Limits...
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.

Show MeSH

Related in: MedlinePlus

Western blot of cTnI, ssTnI, α-MHC, and β-MHC expression in fetal Iso2, fetal Con, adult Iso, and adult Con hearts. Individual protein expression was normalized to GAPDH expression and displayed as a percentage of control. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.0001. n = 9/group, while n = 8 for adult Iso group.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4127273&req=5

fig7: Western blot of cTnI, ssTnI, α-MHC, and β-MHC expression in fetal Iso2, fetal Con, adult Iso, and adult Con hearts. Individual protein expression was normalized to GAPDH expression and displayed as a percentage of control. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.0001. n = 9/group, while n = 8 for adult Iso group.

Mentions: Interestingly, in fetal Iso2 group, ssTnI was repressed compared to normal fetal rats (P < 0.01), while α-MHC was increased according to western blot analysis, while there was no change in cTnI and β-MHC expressions. Besides, the elevated ratios of cTnI/ssTnI and α-MHC/β-MHC level were identified (P < 0.01) (Figure 7). And the same evaluating procedures were performed in adult ones. As same as previous studies, we found that the level of cTnI and α-MHC dropped in adult Iso rats (P < 0.01), while elevating level of ssTnI and β-MHC had been confirmed (P < 0.01) with decreasing ratio of cTnI/ssTnI and α-MHC/β-MHC (P < 0.01) (Figure 7). Besides, RT-PCR was enrolled for mRNA analysis for cTnI, ssTnI, α-MHC, and β-MHC, which had the results consistent with western blot analysis (Figure 8).


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)

Western blot of cTnI, ssTnI, α-MHC, and β-MHC expression in fetal Iso2, fetal Con, adult Iso, and adult Con hearts. Individual protein expression was normalized to GAPDH expression and displayed as a percentage of control. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.0001. n = 9/group, while n = 8 for adult Iso group.
© Copyright Policy
Related In: Results  -  Collection

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

fig7: Western blot of cTnI, ssTnI, α-MHC, and β-MHC expression in fetal Iso2, fetal Con, adult Iso, and adult Con hearts. Individual protein expression was normalized to GAPDH expression and displayed as a percentage of control. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.0001. n = 9/group, while n = 8 for adult Iso group.
Mentions: Interestingly, in fetal Iso2 group, ssTnI was repressed compared to normal fetal rats (P < 0.01), while α-MHC was increased according to western blot analysis, while there was no change in cTnI and β-MHC expressions. Besides, the elevated ratios of cTnI/ssTnI and α-MHC/β-MHC level were identified (P < 0.01) (Figure 7). And the same evaluating procedures were performed in adult ones. As same as previous studies, we found that the level of cTnI and α-MHC dropped in adult Iso rats (P < 0.01), while elevating level of ssTnI and β-MHC had been confirmed (P < 0.01) with decreasing ratio of cTnI/ssTnI and α-MHC/β-MHC (P < 0.01) (Figure 7). Besides, RT-PCR was enrolled for mRNA analysis for cTnI, ssTnI, α-MHC, and β-MHC, which had the results consistent with western blot analysis (Figure 8).

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