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Experimental study on the suppression of sodium nitroprussiate-induced chondrocyte apoptosis by Tougu Xiaotong Capsule (透骨消痛胶囊)-containing serum

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Abstract

Objective

To study the mechanism of action of Tougu Xiaotong Capsule (透骨消痛胶囊, TGXTC) ex vivo in suppressing chondrocyte (CD) apoptosis induced by sodium nitroprussiate (SNP).

Methods

Thirty New Zealand rabbits, 2 months old, were randomized by lottery into five groups, six in each: the blank group treated with saline, the positive control group treated with Zhuanggu Guanjie Pill (壮骨关节丸, 70 mg/kg), and the three experimental groups, EGA, EGB, and EGC, treated with low dose (35 mg/kg), moderate dose (70 mg/kg), and high dose (140 mg/kg) of TGXTC, respectively. All treatments were administered via gastrogavage twice a day for 3 days. Arterial blood was collected from the abdominal aorta and drug or drug metabolites-containing serum was prepared. CDs obtained from knee joints of 16 four-week-old New Zealand rabbits were cultured to the third passage and confirmed by toluidine blue staining. SNP of various final concentrations (0, 0.5, 1.0, and 2.0 mmol/L) was used to induce CD apoptosis, and the dosage-effect relationship of SNP in inducing CD apoptosis was determined. Serum samples from the blank, control, and three dosages of TGXTC-treated rabbits were tested in the CD culture in the presence of SNP. Cell apoptosis was determined by Hoechst 33342 staining, viability of CDs was quantified by MTT, CD apoptosis rate was determined by annexin V-FITC/PI staining, levels of p53 and Bcl-2 mRNA expression in CDs were determined with RT-PCR, and contents of caspase-3 and caspase-9 proteins were determined by colorimetry.

Results

CD apoptosis was induced by SNP at all concentrations tested and in a dose-dependent manner. The SNP concentration of 1 mmol/L and treatment duration of 24 h appeared to be optimal and were selected for the study. Serum samples from the positive control rabbits and from the two higher doses of TGXTC-treated rabbits showed reduction of SNP-induced CD apoptosis, decrease in p53 mRNA expression, inhibition of catalytic activities of caspase-3 and caspase-9, and increase in Bcl-2 mRNA expression when compared with the serum from the blank group (P<0.05).

Conclusion

TGXTC-containing sera antagonized SNP-induced CD apoptosis and the molecular basis for the action was associated with up-regulation of Bcl-2, down-regulation of p53 expression, and inhibition of caspase-3 and caspase-9 catalytic activities.

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Authors and Affiliations

  1. Academy of Integrative Medicine of Fujian University of Traditional Chinese Medicine, Fuzhou, 350108, China

    Xi-hai Li  (李西海), Ming-xia Wu  (吴明霞), Hong-zhi Ye  (叶蕻芝) & Xian-xiang Liu  (刘献祥)

  2. Fujian Provincial Key Lab for Integrative Medical Study on Senile Diseases, Fuzhou, 350108, China

    Wen-lie Chen  (陈文列), Jiu-mao Lin  (林久茂) & Liang-pu Zheng  (郑良朴)

Authors
  1. Xi-hai Li  (李西海)
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  2. Ming-xia Wu  (吴明霞)
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  3. Hong-zhi Ye  (叶蕻芝)
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  4. Wen-lie Chen  (陈文列)
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  5. Jiu-mao Lin  (林久茂)
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  6. Liang-pu Zheng  (郑良朴)
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  7. Xian-xiang Liu  (刘献祥)
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Corresponding author

Correspondence to Ming-xia Wu  (吴明霞).

Additional information

Supported by the National Natural Science Foundation of China (No. 81072826), Major Project Foundation of Fujian Provincial Administration of Science and Technology (No. 2009Y0029), and the Developmental Fund of CHEN Keji Integrative Medicine (No. CKJ2010013)

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Li, Xh., Wu, Mx., Ye, Hz. et al. Experimental study on the suppression of sodium nitroprussiate-induced chondrocyte apoptosis by Tougu Xiaotong Capsule (透骨消痛胶囊)-containing serum. Chin. J. Integr. Med. 17, 436–443 (2011). https://doi.org/10.1007/s11655-011-0751-x

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  • DOI: https://doi.org/10.1007/s11655-011-0751-x

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