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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (3): 232-236.doi: 10.3736/jcim20090307

• Original Experimental Research • Previous Articles     Next Articles

Effects of tetramethylpyrazine on angiotensin Ⅱ-induced proliferation and type Ⅰ collagen synthesis of rat cardiac fibroblasts

Dong-mei Zhang , Ying Qin , Xi-ying Lu , Ai-ming Wu , Ling-qun Zhu , Shuo-ren Wang , Liang-duo Jiang   

  1. Key Laboratory of Chinese Internal Medicine of the Ministry of Education, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China
  • Received:2008-08-18 Accepted:2008-10-13 Online:2009-03-20 Published:2009-03-15

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Objective

To observe the effects of tetramethylpyrazine (TMP) on the proliferation and type Ⅰ collagen synthesis of rat cardiac fibroblasts (CFBs) induced by angiotensin Ⅱ (Ang Ⅱ), and to explore the mechanism of TMP in treating myocardial fibrosis.
Methods

CFBs were isolated from neonatal rats, and the fourth-passage CFBs were used in the entire test and were stimulated by 0.1 μmol/L Ang Ⅱ in vivo. The CFB proliferation was measured by methyl thiazolyl tetrazolium (MTT) assay. Type Ⅰ collagen in the cell culture supernatant was measured by enzyme-linked immunosorbent assay. The expression of mRNA of type Ⅰ collagen was semi-quantitatively measured by reverse transcription-polymerase chain reaction (RT-PCR).
Results

(1) In MTT assay, the optical density of CFBs cultured with 0.1 μmol/L Ang Ⅱ was higher than that of the blank control cultured with 2% fetal bovine serum-Dulbecco’s modified Eagle’s medium (FBS-DMEM). The difference was statistically significant (P<0.05). Both optical densities of CFBs cultured with 0.1 μmol/L Ang Ⅱ plus 800 μg/mL TMP and 0.1 μmol/L Ang Ⅱ plus 600 μg/mL TMP were lower than that of CFBs cultured with 0.1 μmol/L Ang Ⅱ, but only the difference between 0.1 μmol/L AngⅡ plus 800 μg/mL TMP group and 0.1 μmol/L Ang Ⅱ group was significant (P<0.05). (2) The content of type Ⅰ collagen secreted by CFBs cultured with 0.1 μmol/L Ang Ⅱ was higher than that with 2% FBS-DMEM (P<0.01). The content of type Ⅰ collagen secreted by CFBs cultured with 0.1 μmol/L Ang Ⅱ plus 800 μg/mL TMP was lower than that with 0.1 μmol/L Ang Ⅱ (P<0.05). (3) The level of type Ⅰ collagen mRNA in 0.1 μmol/L Ang Ⅱ group was higher than that in blank control group, and lower than that in 0.1 μmol/L Ang Ⅱ plus 800 μg/mL TMP group. Both the differences between 0.1 μmol/L Ang Ⅱ group and the blank control group and between 0.1 μmol/L Ang Ⅱ group and 0.1 μmol/L Ang Ⅱ plus 800 μg/mL TMP group were statistically significant (P<0.05).
Conclusion

TMP can not only inhibit the proliferation of CFBs, but also decrease the secretion and the mRNA expression level of collagen Ⅰ in cultured CFBs of rat which are increased by Ang Ⅱ.

Key words: Tetramethylpyrazine, Cardiac fibroblasts, Type I collagen, Angiotensin H, Rats

Table 1

Effects of TMP on proliferation of rat CFBs induced by Ang Ⅱ（x±s）"

Group n Optical density
Blank control 6 0.109±0.014
0.1 μmol/L Ang Ⅱ 6 0.133±0.011*
0.1 μmol/L Ang Ⅱ plus 800 μg/mL TMP 6 0.095±0.018△
0.1 μmol/L Ang Ⅱ plus 600 μg/mL TMP 6 0.108±0.015

Table 2

Effects of TMP on secretion of type Ⅰ collagen of rat CFBs induced by Ang Ⅱ（x±s, ng/mg）"

Group n Content of type
Ⅰ collagen
Blank control 6 170.95±13.59
0.1 μmol/L AngⅡ 6 215.61±14.95**
0.1 μmol/L Ang Ⅱ plus 800 μg/mL TMP 6 183.09±15.13△

Table 3

Effects of TMP on expression of type Ⅰ collagen mRNA of rat CFBs induced by Ang Ⅱ（x±s）"

Group n Expression level of type Ⅰ collagen mRNA
Blank control 5 0.975±0.049
0.1 μmol/L Ang Ⅱ 5 1.216±0.096*
0.1 μmol/L Ang Ⅱ plus 800 μg/mL TMP 5 0.966±0.104△
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