Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (1): 20-25.doi: 10.3969/j.issn.2095-4344.2146

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Effect of Yougui Decoction on autophagy and fate of bone marrow mesenchymal stem cells in rats with glucocorticoid-associated femoral head necrosis

Liu Xin1 , Du Bin1 , Gao Lili1 , Sun Guangquan1 , Wang Xu2 , Yuan Peng2 , Lin Xuanye2
  

  1. 1 Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital of Chinese Medicine, Nanjing 210029, Jiangsu Province, China; 2 First Clinical School of Nanjing University of Chinese Medicine, Nanjing 210023, Jiangsu Province, China
  • Received:2020-01-11 Revised:2020-01-15 Accepted:2020-02-23 Online:2021-01-08 Published:2020-09-19
  • Contact: Du Bin, MD, Chief physician, Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital of Chinese Medicine, Nanjing 210029, Jiangsu Province, China
  • About author:Liu Xin, Master, Physician, Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital of Chinese Medicine, Nanjing 210029, Jiangsu Province, China

Abstract: BACKGROUND: Yougui Decoction is an empirical prescription for the treatment of glucocorticoid-associated femoral head necrosis. Literature has shown that the pathogenesis of glucocorticoid-associated femoral head necrosis is associated with glucocorticoid-induced autophagy down-regulation and fate change in bone marrow mesenchymal stem cells. 
OBJECTIVE: To investigate the effect of Yougui Decoction on autophagy and fate of bone marrow mesenchymal stem cells in model rats of glucocorticoid-associated femoral head necrosis. 
METHODS: We used Escherichia coli endotoxin combined with high-dose dexamethasone to make the rat models of early femoral head necrosis. Forty SHR rats were randomly divided into five groups: blank control group, model group, high-dose Yougui Decoction group, medium-dose Yougui Decoction group and low-dose Yougui Decoction group. After 6 weeks of intervention, medullary cavity tissue of the rat proximal femur was taken for hematoxylin-eosin staining and immunohistochemical staining of autophagy proteins LC3 II, P53 and beclin-1. After culture and induction of bone marrow mesenchymal stem cells, alizarin red staining, bone alkaline phosphatase quantification, oil red staining and MTT determination were performed and western blot assay was used to quantitatively measure the expression of LC3 II, P53 and beclin-1 proteins. 
RESULTS AND CONCLUSION: (1) Immunohistochemistry and western blot assay results showed that Yougui Decoction significantly increased autophagy protein LC3 II, P53 and beclin 1 expression in a dose-dependent manner. (2) Alizarin red staining, oil red staining and bone alkaline phosphatase quantification suggested that Yougui Decoction could significantly interfere with the fate of bone marrow mesenchymal stem cells, up-regulate their osteogenic differentiation and down-regulate their adipogenic differentiation in a dose-dependent manner. (3) MTT results suggested that Yougui Decoction significantly improved the proliferation ability of bone marrow mesenchymal stem cells, but had no significant differences in different doses. (4) To conclude, Yougui Decoction can significantly improve the autophagy expression, change the cell fate, up-regulate osteogenic differentiation and down-regulate adipogenic differentiation of bone marrow mesenchymal stem cells in the rat models of glucocorticoid-associated femoral head necrosis, which provides certain basis for elucidating the mechanism of Yougui Decoction in treating glucocorticoid-associated femoral head necrosis.

Key words: Yougui Decoction, glucocorticoid-associated femoral head necrosis, bone marrow, mesenchymal stem cells, autophagy, cell fate, rat

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