Limits...
Circulating miRNAs reflect early myocardial injury and recovery after heart transplantation.

Wang E, Nie Y, Zhao Q, Wang W, Huang J, Liao Z, Zhang H, Hu S, Zheng Z - J Cardiothorac Surg (2013)

Bottom Line: Correlations between miRNAs and cTnI were statistically significant (p < 0.05), especially for miR-133b (R2 = 0.813, p < 0.001).Obviously, miR-133b had a better correlation than cTnI.And miR-133b may have advantages over cTnI in forecasting graft dysfunction and recovery of patients after operation.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100037, People's Republic of China.

ABSTRACT

Background: MicroRNAs (miRNAs) are short, single-stranded and non-coding RNAs, freely circulating in human plasma and correlating with vary pathologies. In this study, we monitored early myocardial injury and recovery after heart transplantation by detecting levels of circulating muscle-specific miR-133a, miR-133b and miR-208a.

Methods: 7 consecutive patients underwent heart transplantation in Fuwai hospital and 14 healthy controls were included in our study. Peripheral vein blood was drawn from patients on the day just after transplantation (day 0), the 1st, 2nd, 3rd, 7th and 14th day after transplantation respectively. Serum from peripheral blood was obtained for cardiac troponin I (cTnI) measurement. Plasma was centrifuged from peripheral blood for measuring miR-133a, miR-133b and miR-208a by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The plasma concentration of miRNAs were calculated by absolute quantification method. The sensitivity and specificity of circulating miRNAs were revealed by receiver operating characteristic curve (ROC) analysis. Correlations between miRNAs and cTnI / perioperative parameters were analyzed.

Results: Similar to cTnI, miR-133a, miR-133b and miR-208a all showed dynamic changes from high to low levels early after operation. The Sensitivity and specificity of miRNAs were: miR-133a (85.7%,100%), miR-208a (100%,100%), and miR-133b (90%,100%). Correlations between miRNAs and cTnI were statistically significant (p < 0.05), especially for miR-133b (R2 = 0.813, p < 0.001). MiR-133b from Day 0-Day 2 (r > 0.98, p < 0.01), and cTnI from Day 1- Day 3 (r > 0.86, p < 0.05) had strong correlations with bypass time, particularly parallel bypass time. Obviously, miR-133b had a better correlation than cTnI. Circulating miR-133b correlated well with parameters of heart function such as central venous pressure (CVP), pulmonary capillary wedge pressure (PCWP), cardiac output (CO) and inotrope support, while cTnI only correlated with 3 of the 4 parameters mentioned above. MiR-133b also had strong correlations with ventilation time (r > 0.99, p < 0.001) and length of ICU stay (r > 0.92, p < 0.05), both of which reflected the recovery after operation. The correlation coefficients of miR-133b were also higher than that of cTnI.

Conclusions: The dynamic change in circulating muscle-specific miRNAs, especially miR-133b can reflect early myocardial injury after heart transplantation. And miR-133b may have advantages over cTnI in forecasting graft dysfunction and recovery of patients after operation.

Show MeSH

Related in: MedlinePlus

Standard curves of miR-133a, miR-208a, and miR-133b for TaqMan qRT-PCR assays. Standard curves were generated for each miRNAs assay using a dilution series of known input amounts of synthetic miRNAs oligonucleotide corresponding to the target of the assay (Table 1). Data points not on the trend line are those for which the measured Ct was interpreted to be below the linear range of the assay, and were not used for derivation of the trend line. A, Standard curve of miR-133a; B, Standard curve of miR-208a. C, Standard curve of miR-133b.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3716980&req=5

Figure 1: Standard curves of miR-133a, miR-208a, and miR-133b for TaqMan qRT-PCR assays. Standard curves were generated for each miRNAs assay using a dilution series of known input amounts of synthetic miRNAs oligonucleotide corresponding to the target of the assay (Table 1). Data points not on the trend line are those for which the measured Ct was interpreted to be below the linear range of the assay, and were not used for derivation of the trend line. A, Standard curve of miR-133a; B, Standard curve of miR-208a. C, Standard curve of miR-133b.

Mentions: Synthetic single-stranded RNA oligonucleotides miR-133a, miR-208a, and miR-133b (miRBase Release v.19.0) were purchased from Shanghai GenePharma, China. Sequence information was provided in Table 2. Synthetic miRNAs were input into the RT reaction according to range of copies from reference [3,20]. The standard curves for miR133a, miR-208a, and miR-133b were plotted by Ct values versus copy number of the synthetic miRNAs as shown in Figure 1. Copies of endogenous miRNAs in plasma were then approximated according to their Ct values and the standard curve. A Normalization Factor was calculated by the Ct values of the three synthetic spiked-in C. elegans miRNAs [3,20]. Then the copies of a given miRNAs in each sample (calculated by the standard curves described earlier) were multiplied by the normalization factor corresponding to the sample to obtain a normalized copy number [3,20].


Circulating miRNAs reflect early myocardial injury and recovery after heart transplantation.

Wang E, Nie Y, Zhao Q, Wang W, Huang J, Liao Z, Zhang H, Hu S, Zheng Z - J Cardiothorac Surg (2013)

Standard curves of miR-133a, miR-208a, and miR-133b for TaqMan qRT-PCR assays. Standard curves were generated for each miRNAs assay using a dilution series of known input amounts of synthetic miRNAs oligonucleotide corresponding to the target of the assay (Table 1). Data points not on the trend line are those for which the measured Ct was interpreted to be below the linear range of the assay, and were not used for derivation of the trend line. A, Standard curve of miR-133a; B, Standard curve of miR-208a. C, Standard curve of miR-133b.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3716980&req=5

Figure 1: Standard curves of miR-133a, miR-208a, and miR-133b for TaqMan qRT-PCR assays. Standard curves were generated for each miRNAs assay using a dilution series of known input amounts of synthetic miRNAs oligonucleotide corresponding to the target of the assay (Table 1). Data points not on the trend line are those for which the measured Ct was interpreted to be below the linear range of the assay, and were not used for derivation of the trend line. A, Standard curve of miR-133a; B, Standard curve of miR-208a. C, Standard curve of miR-133b.
Mentions: Synthetic single-stranded RNA oligonucleotides miR-133a, miR-208a, and miR-133b (miRBase Release v.19.0) were purchased from Shanghai GenePharma, China. Sequence information was provided in Table 2. Synthetic miRNAs were input into the RT reaction according to range of copies from reference [3,20]. The standard curves for miR133a, miR-208a, and miR-133b were plotted by Ct values versus copy number of the synthetic miRNAs as shown in Figure 1. Copies of endogenous miRNAs in plasma were then approximated according to their Ct values and the standard curve. A Normalization Factor was calculated by the Ct values of the three synthetic spiked-in C. elegans miRNAs [3,20]. Then the copies of a given miRNAs in each sample (calculated by the standard curves described earlier) were multiplied by the normalization factor corresponding to the sample to obtain a normalized copy number [3,20].

Bottom Line: Correlations between miRNAs and cTnI were statistically significant (p < 0.05), especially for miR-133b (R2 = 0.813, p < 0.001).Obviously, miR-133b had a better correlation than cTnI.And miR-133b may have advantages over cTnI in forecasting graft dysfunction and recovery of patients after operation.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100037, People's Republic of China.

ABSTRACT

Background: MicroRNAs (miRNAs) are short, single-stranded and non-coding RNAs, freely circulating in human plasma and correlating with vary pathologies. In this study, we monitored early myocardial injury and recovery after heart transplantation by detecting levels of circulating muscle-specific miR-133a, miR-133b and miR-208a.

Methods: 7 consecutive patients underwent heart transplantation in Fuwai hospital and 14 healthy controls were included in our study. Peripheral vein blood was drawn from patients on the day just after transplantation (day 0), the 1st, 2nd, 3rd, 7th and 14th day after transplantation respectively. Serum from peripheral blood was obtained for cardiac troponin I (cTnI) measurement. Plasma was centrifuged from peripheral blood for measuring miR-133a, miR-133b and miR-208a by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The plasma concentration of miRNAs were calculated by absolute quantification method. The sensitivity and specificity of circulating miRNAs were revealed by receiver operating characteristic curve (ROC) analysis. Correlations between miRNAs and cTnI / perioperative parameters were analyzed.

Results: Similar to cTnI, miR-133a, miR-133b and miR-208a all showed dynamic changes from high to low levels early after operation. The Sensitivity and specificity of miRNAs were: miR-133a (85.7%,100%), miR-208a (100%,100%), and miR-133b (90%,100%). Correlations between miRNAs and cTnI were statistically significant (p < 0.05), especially for miR-133b (R2 = 0.813, p < 0.001). MiR-133b from Day 0-Day 2 (r > 0.98, p < 0.01), and cTnI from Day 1- Day 3 (r > 0.86, p < 0.05) had strong correlations with bypass time, particularly parallel bypass time. Obviously, miR-133b had a better correlation than cTnI. Circulating miR-133b correlated well with parameters of heart function such as central venous pressure (CVP), pulmonary capillary wedge pressure (PCWP), cardiac output (CO) and inotrope support, while cTnI only correlated with 3 of the 4 parameters mentioned above. MiR-133b also had strong correlations with ventilation time (r > 0.99, p < 0.001) and length of ICU stay (r > 0.92, p < 0.05), both of which reflected the recovery after operation. The correlation coefficients of miR-133b were also higher than that of cTnI.

Conclusions: The dynamic change in circulating muscle-specific miRNAs, especially miR-133b can reflect early myocardial injury after heart transplantation. And miR-133b may have advantages over cTnI in forecasting graft dysfunction and recovery of patients after operation.

Show MeSH
Related in: MedlinePlus