Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (3): 448-457.DOI: 10.19852/j.cnki.jtcm.20240402.004
• Original Articles • Previous Articles Next Articles
ZHANG Zeyu, JIA Zhuangzhuang, SONG Yuwei, ZHANG Xuan, WANG Ci, WANG Shuai, ZHANG Peipei, REN Qiuan, WANG Xianliang(), MAO Jingyuan()
Received:
2023-01-16
Accepted:
2023-05-25
Online:
2024-04-02
Published:
2024-04-02
Contact:
WANG Xianliang,MAO Jingyuan
About author:
Prof. MAO Jingyuan, Department of First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion, Tianjin 300381, China. jymao@126.com Telephone: +86-13920007825; +86-13820498886Supported by:
ZHANG Zeyu, JIA Zhuangzhuang, SONG Yuwei, ZHANG Xuan, WANG Ci, WANG Shuai, ZHANG Peipei, REN Qiuan, WANG Xianliang, MAO Jingyuan. Optimized new Shengmai powder (优化新生脉散方) inhibits myocardial fibrosis in heart failure by regulating the rat sarcoma/rapidly accelerated fibrosarcoma/mitogen-activated protein kinase kinase/extracellular regulated protein kinases signaling pathway[J]. Journal of Traditional Chinese Medicine, 2024, 44(3): 448-457.
Primer name | Sequence (5’-3’) | Size (bp) | |
---|---|---|---|
COL Ⅰ | F | GAG AGG TGA ACA AGG TCC CG | 153 |
R | AAA CCT CTC TCG CCT CTT GC | ||
COL Ⅲ | F | AAG GCT GCA AGA TGG ATG CT | 95 |
R | GTG CTT ACG TGG GAC AGT CA | ||
α-SMA | F | CTATGAGGGCTATGCCTTGCC | 122 |
R | GCTCAGCAGTAGTAACGAAGGA | ||
c-Fos | F | GTCTTCCTTTGTCTTCACCTACCC | 384 |
R | CCCTGCCTTCTCTGACTGCT | ||
β-actin | F | GTCGTACCACTGGCATTGTG | 180 |
R | TCTCAGCTGTGGTGGTGAAG |
Table 1 Primer sequence table
Primer name | Sequence (5’-3’) | Size (bp) | |
---|---|---|---|
COL Ⅰ | F | GAG AGG TGA ACA AGG TCC CG | 153 |
R | AAA CCT CTC TCG CCT CTT GC | ||
COL Ⅲ | F | AAG GCT GCA AGA TGG ATG CT | 95 |
R | GTG CTT ACG TGG GAC AGT CA | ||
α-SMA | F | CTATGAGGGCTATGCCTTGCC | 122 |
R | GCTCAGCAGTAGTAACGAAGGA | ||
c-Fos | F | GTCTTCCTTTGTCTTCACCTACCC | 384 |
R | CCCTGCCTTCTCTGACTGCT | ||
β-actin | F | GTCGTACCACTGGCATTGTG | 180 |
R | TCTCAGCTGTGGTGGTGAAG |
Figure 1 Comparison of cardiac function and organ coefficient in rats of each group A: typical echocardiographic image. A1-A6: the echocardiographic profile of sham (A1), model (A2), ONSMP-L (A3), ONSMP-M (A4), ONSMP-H (A5), and ENP (A6); B: statistic of LVEF of rats in each group; C: Statistic of LVFS of rats in each group; D: Statistic of HMI of rats in each group; E: statistic of LVMI of rats in each group. Sham group: sham operated; model group: ligation of the left anterior descending coronary artery without treatment; ONSMP-L: ligation of the left anterior descending coronary artery rats treated with ONSMP intragastrically at a daily dose of 4.05 g crude drug/kg for 4 weeks; ONSMP-M: ligation of the left anterior descending coronary artery rats treated with ONSMP intragastrically at a daily dose of 8.1 g crude drug/kg for 4 weeks; ONSMP-H: ligation of the left anterior descending coronary artery rats treated with ONSMP intragastrically at a daily dose of 16.2 g crude drug/kg for 4 weeks; ENP: ligation of the left anterior descending coronary artery rats treated with ENP intragastrically at a daily dose of 0.9 g/kg for 4 weeks. ONSMP: Optimized New Shengmai powder; ENP: enalapril; LVEF: left ventricular ejection fractions; LVFS: left ventricular fractional shortening; HWI: heart weight index; LVMI: left ventricular mass index. Data represent the mean ± standard deviation using one-way analysis of variance (n = 6). Compared with the sham group, aP < 0.01; compared with the model group, bP < 0.01; compared with the ONSMP-L group, cP < 0.01; compared with the ONSMP-M group, dP < 0.01.
Figure 2 Pathological section results of myocardial tissue of rats in each group A: typical myocardial HE-stained section image. A1-A6: HE-stained section images of sham (A1), model (A2), ONSMP-L (A3), ONSMP-M (A4), ONSMP-H (A5), and ENP (A6) (× 1); A7-A12: HE-stained section images of sham (A7), model (A8), ONSMP-L (A9), ONSMP-M (A10), ONSMP-H (A11), and ENP (A12) (× 200). B: Typical myocardial masson-stained section image. B1-B6: masson-stained section images of sham (B1), model (B2), ONSMP-L (B3), ONSMP-M (B4), ONSMP-H (B5), and ENP (B6) (× 1); B7-B12: masson-stained section images of sham (B7), model (B8), ONSMP-L (B9), ONSMP-M (B10), ONSMP-H (B11), and ENP (B12) (× 200). C: statistics of CVF of rats in each group. Sham group: sham operated; model group: ligation of the left anterior descending coronary artery without treatment; ONSMP-L: ligation of the left anterior descending coronary artery rats treated with ONSMP intragastrically at a daily dose of 4.05 g crude drug/kg for 4 weeks; ONSMP-M: ligation of the left anterior descending coronary artery rats treated with ONSMP intragastrically at a daily dose of 8.1 g crude drug/kg for 4 weeks; ONSMP-H: ligation of the left anterior descending coronary artery rats treated with ONSMP intragastrically at a daily dose of 16.2 g crude drug/kg for 4 weeks; ENP: ligation of the left anterior descending coronary artery rats treated with ENP intragastrically at a daily dose of 0.9 g/kg for 4 weeks. ONSMP: Optimized New Shengmai powder; ENP: enalapril; CVF: collagen volume fraction. Data represent the mean ± standard deviation using one-way analysis of variance (n = 3). Compared with the sham group, aP < 0.01; compared with the model group, bP < 0.01; compared with the ONSMP-L group, cP < 0.01; compared with the ONSMP-M group, dP < 0.01.
Group | n | COL Ⅰ | COL Ⅲ |
---|---|---|---|
Sham | 6 | 34.5±1.4 | 20.4±1.6 |
Model | 6 | 44.3±1.3a | 27.1±1.3a |
ONSMP-L | 6 | 40.5±1.7c | 24.7±1.5b |
ONSMP-M | 6 | 39.5±1.8c | 23.1±1.1c |
ONSMP-H | 6 | 36.1±1.8cef | 22.4±0.8cd |
Table 2 Content of COL Ⅰ and COL Ⅲ in myocardial tissue of rats in each group (μg/L, $\bar{x} \pm s$)
Group | n | COL Ⅰ | COL Ⅲ |
---|---|---|---|
Sham | 6 | 34.5±1.4 | 20.4±1.6 |
Model | 6 | 44.3±1.3a | 27.1±1.3a |
ONSMP-L | 6 | 40.5±1.7c | 24.7±1.5b |
ONSMP-M | 6 | 39.5±1.8c | 23.1±1.1c |
ONSMP-H | 6 | 36.1±1.8cef | 22.4±0.8cd |
Group | n | COL Ⅰ/β-actin | COL Ⅲ/β-actin | α-SMA/β-actin | c-Fos/β-actin |
---|---|---|---|---|---|
Sham | 3 | 1.04±0.29 | 1.04±0.28 | 1.01±0.15 | 1.08±0.40 |
Model | 3 | 7.69±0.50a | 7.03±0.74a | 7.36±1.62a | 6.45±0.70a |
ONSMP-L | 3 | 4.85±0.42b | 4.12±0.22b | 5.02±0.70b | 2.92±0.16b |
ONSMP-M | 3 | 2.73±0.33bc | 2.19±0.30bc | 3.26±0.68bc | 2.05±0.33bc |
ONSMP-H | 3 | 1.32±0.11bcd | 1.30±0.05bcd | 1.79±0.32bcd | 1.35±0.05bcd |
Table 3 mRNA of COL Ⅰ, COL Ⅲ, α-SMA, and c-Fos in myocardial tissue of rats in each group ($\bar{x} \pm s$)
Group | n | COL Ⅰ/β-actin | COL Ⅲ/β-actin | α-SMA/β-actin | c-Fos/β-actin |
---|---|---|---|---|---|
Sham | 3 | 1.04±0.29 | 1.04±0.28 | 1.01±0.15 | 1.08±0.40 |
Model | 3 | 7.69±0.50a | 7.03±0.74a | 7.36±1.62a | 6.45±0.70a |
ONSMP-L | 3 | 4.85±0.42b | 4.12±0.22b | 5.02±0.70b | 2.92±0.16b |
ONSMP-M | 3 | 2.73±0.33bc | 2.19±0.30bc | 3.26±0.68bc | 2.05±0.33bc |
ONSMP-H | 3 | 1.32±0.11bcd | 1.30±0.05bcd | 1.79±0.32bcd | 1.35±0.05bcd |
Figure 3 ONSMP inhibited the RAS/RAF/MEK/ERK pathway and downstream protein expression A: Western blotting representative images of p-RAS, p-RAF, p-MEK1/2, p-ERK1/2, p-ELK1, p-c-Fos, COL I, COL Ⅲ, and α-SMA. B: protein expression levels of p-RAS; C: protein expression levels of p-RAF; D: protein expression levels of p-MEK1/2; E: protein expression levels of p-ERK1/2; F: protein expression levels of p-ELK1; G: protein expression levels of p-c-Fos; H: protein expression levels of COL I; I: protein expression levels of COL Ⅲ; J: protein expression levels of α-SMA. Sham group: sham operated; model group: ligation of the left anterior descending coronary artery without treatment; ONSMP-L: ligation of the left anterior descending coronary artery rats treated with ONSMP intragastrically at a daily dose of 4.05 g crude drug/kg for 4 weeks; ONSMP-M: ligation of the left anterior descending coronary artery rats treated with ONSMP intragastrically at a daily dose of 8.1 g crude drug/kg for 4 weeks; ONSMP-H: ligation of the left anterior descending coronary artery rats treated with ONSMP intragastrically at a daily dose of 16.2 g crude drug/kg for 4 weeks; ENP: ligation of the left anterior descending coronary artery rats treated with ENP intragastrically at a daily dose of 0.9 g/kg for 4 weeks. p-RAS: phosphor-rat sarcoma; p-RAF: phosphor-rapidly accelerated fibrosarcoma; p-MEK1/2: phosphor-mitogen-activated protein kinase kinase 1/2; p-ERK1/2: phosphor-extracellular regulated protein kinases; p-ELK1: phosphor-ETS-like-1 transcription factor; p-c-Fos: phosphor-c-Fos proto-oncogene; COL I: collagen I; COL Ⅲ: collagen Ⅲ; α-SMA: α-smooth muscle actin; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; ONSMP: optimized New Shengmai powder; ENP: enalapril. Data represent the mean ± standard deviation using one-way analysis of variance (n = 3). Compared with the sham group, aP < 0.01; compared with the model group, bP < 0.01; compared with the ONSMP-L group, cP < 0.05, dP < 0.01; compared with the ONSMP-M group, eP < 0.05.
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