Development of an efficient mice model of cancer‐associated cardiac cachexia

Figures

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  FIGURE 1. Cardiac function of mice with their respective control groups evaluated. (A) Representative echocardiography and assessment of control mice and CT26‐S mice (n = 8). (B) Representative echocardiography and assessment of control mice and CT26‐I mice (n = 8). (C) Representative diagram of wasting and cardiac atrophy in CT26‐I mice. (D) Epididymal fat weight/tibial length, weight/body weight, heart weight/body weight, and heart weight/tibial length of CT26‐I mice and control mice (n = 5). (E) Survival proportions of CT26‐I mice and control mice (n = 5). (F) Left ventricular mass in CT26‐I mice and control mice (n = 5). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns p > 0.05 compared to control group.

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  FIGURE 2. Cardiac morphological pathology of CT26‐I mice and cardiomyocyte size isolated from mice. (A) Left: Representative image of Masson's staining of heart tissue from the indicated mice (n = 4). Scale bars, 50 μm and 20 μm. Middle: Representative image of WGA staining of heart from the indicated mice(n = 4). Scale bars, 75 μm and 25 μm. Right: Representative image of hematoxylin–eosin (HE) staining of heart from the indicated mice (n = 4). Scale bars, 50 μm and 20 μm. (B) Quantitative analysis of interstitial fibrosis in cardiac tissue by Masson's staining. (C) Quantitative analysis of cardiomyocytes area by WGA staining. (D) Quantitative analysis of transverse myocyte diameter by HE staining. Each group has consisted of independent mice (n = 4). (E) The cardiomyocytes of CT26‐I mice contain sparse myofibrils, partially broken myofibrils, locally lysed myofibers, and disorderly sarcomeres. Magnification: 9700× and 37 000×. (F) Representative plot of cardiomyocyte population of control mice and CT26‐I mice. (G) Enrichment map of cardiomyocyte population in CT26‐I mice compared to control mice. (H–J) Cardiomyocyte size is assessed by forward scattered light signals (FSC‐H, FSC‐width,…

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  FIGURE 3. ECG monitoring of mice and cardiomyocyte contractility. (A) Resting heart rate in CT26‐I mice versus control mice. (B) Duration of PQ in CT26‐I mice versus control mice. (C) Duration of QRS in CT26‐I mice versus control mice. (D) QT duration of CT26‐I mice versus control mice. (E) ST duration of CT26‐I mice versus control mice. (F) QCT duration of CT26‐I mice versus control mice (n = 4). (G) Representative ECGs of CT26‐I mice and control mice. Each group consisted of four independent mice. (H) Average sarcomere shortening traces were compared. Shortening is shown normalized to resting length; the negative deflection represents contraction in response to electrical stimulation followed by relaxation back to the resting length. (n = 4 mice, n = 65 cardiomyocytes examined over four independent experiments) (I) Resting sarcomere length per cardiomyocytes are shown as circles. (J) Sarcomere peak shortening. (K) Pooled data of contraction velocity and relaxation velocity, and pooled data of contraction time and relaxation time to peak (50%). The box bounds represent the 25th and 75th percentiles; the middle line shows the median; the whiskers show the minimum and maximum; and…

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  FIGURE 4. Upregulation of cardiomyocyte‐specific E3 ubiquitination ligase levels and alterations of cardiac myosin isoform in cancer‐associated cardiac cachexia (CACC) mice. (A) Cardiac sarcomere protein transcription levels in CT26‐I mice (n = 4) using quantitative polymerase chain reaction (qPCR). (B) Cardiac sarcomere protein transcription levels in LLC‐T mice (n = 4) using qPCR. (C) Quantification of relative expression of Myh6/Myh7 in CACC mice (n = 3) by Western blotting (WB). (D) WB map of Myh6 and Myh7 in heart of CACC mice (n = 3) compared to control mice. (E) Quantification of the respective expression of Myh6 and Myh7 in CACC mice by WB. (F) Representative WB map of cardiac Ub‐protein conjugates in heart of CACC mice (n = 3) compared to control mice. (G) Quantification of cardiac Ub‐protein conjugates in heart of CACC mice compared to control mice. (H) WB map of MuRF‐1 and Atrogin‐1 in the hearts of CACC mice (n = 3) compared to control mice. (I) Quantification of MuRF‐1 and Atrogin‐1 in the hearts of CACC mice compared to control mice. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 compared to control group.