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. 2017 Jun;13(6):3013-3020.
doi: 10.3892/etm.2017.4349. Epub 2017 Apr 18.

Protective effect of Shenmai injection on knee articular cartilage of osteoarthritic rabbits and IL-1β-stimulated human chondrocytes

Nan Yao  1   2 , Neng Chen  1   3 , Xuemeng Xu  1 , Dongmei Sun  2 , Wengang Liu  1 , Gang Li  4 , Xiaoli Bi  2 , Sumei Li  2 , Zhao Chen  2 , Guocai Chen  1   3 , Haining Gan  2
Affiliations

Protective effect of Shenmai injection on knee articular cartilage of osteoarthritic rabbits and IL-1β-stimulated human chondrocytes

Nan Yao et al. Exp Ther Med. 2017 Jun.

Abstract

Shenmai injection (SMI) has been widely used as a therapy to treat a number of diseases. However, its anti-osteoarthritic properties have not yet been fully investigated. In the present study, the protective effect of SMI on knee articular cartilage of anterior cruciate ligament transected rabbits and interleukin-1β (IL-1β)-stimulated human chondrocytes was investigated. For the in vivo study, knee osteoarthritis (KOA) was induced in female New Zealand white rabbits by anterior cruciate ligament transection (ACLT) in the knee of right hind limb. Rabbits either underwent sham surgery or ACLT surgery. Out of the rabbits receiving ACLT surgery, half of the rabbits received one 0.3 ml Shenmai intra-articular injection in the knee per week for four weeks, following ACLT surgery. The other rabbits received the same volume of normal saline solution. The cartilage was subsequently collected for histological evaluation. For the in vitro study, cultured human chondrocytes were treated with 10 ng/ml IL-1β in the presence or absence of 5 and 2% (v/v) SMI for 24 h. Nitric oxide (NO) and prostaglandin E2 (PGE2) levels in cell culture supernatant were assessed using a Griess reaction and ELISA respectively. The mRNA expression of cyclooxgenase-2 (COX-2), inducible nitric oxide synthase (iNOS), matrix metalloproteinase (MMP)-1, MMP-13 and tissue inhibitors of metalloproteinase-1 (TIMP-1) in chondrocytes were detected by reverse transcription-quantitative polymerase chain reaction. The results of the current study revealed that treatment with SMI ameliorated cartilage degradation in the ACLT rabbit model, and decreased levels of NO and PGE2. Furthermore, treatment with SMI decreased levels of COX-2, iNOS, MMP-1 and MMP-13 mRNA expression and increased TIMP-1 mRNA expression in IL-1β-stimulated human chondrocytes. These results indicate that SMI suppresses inflammation and ameliorated cartilage degradation, making it a potential and promising therapeutic option to treat KOA.

Keywords: Shenmai injection; anterior cruciate ligament transection; cartilage; chondrocytes; knee osteoarthritis.

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Figures

Figure 1.
Figure 1.
Chemical profile of Shenmai injection analyzed by high performance lipid chromatography.
Figure 2.
Figure 2.
H&E and Safranin O-Fast Green staining of articular cartilage in different groups (magnification ×100). In H&E staining, the nucleus was stained purple-blue and cytoplasm was stained red. In Safranin O-Fast Green staining, cartilage was stained red and subchondral bone was stained blue. H&E, hematoxylin and eosin.
Figure 3.
Figure 3.
Effect of SMI on relative cell viability. The results are presented as mean ± standard deviation obtained from the 6 wells in each group. **P<0.01 vs. Control group. SMI, shenmai injection.
Figure 4.
Figure 4.
Effect of SMI on NO and PGE2 levels in human chondrocytes. Results are presented as mean ± standard deviation obtained from 3 wells in each group. ##P<0.01 vs. Control group; *P<0.05, **P<0.01 vs. IL-1β group. SMI, Shenmai injection; NO, nitric oxide; PGE2, prostaglandin; IL-1β, interleukin-1β.
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