Ying Hao1, Yang Wu2, Shanglong Wang3, Chungguo Wang4, Sihao Qu5, Li Li6, Guohua Yu7, Zimin Liu8, Zhen Zhao9, Pengcheng Fan10, Zengliang Zhang11, Yuanyuan Shi12. 1. School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 102488, China; Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY, 10065, USA. Electronic address: haoying_ibp@163.com. 2. School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 102488, China. Electronic address: 977672064@qq.com. 3. Chenland Nutritionals, Inc, Irvine, CA, 92614, USA. Electronic address: mwang@chenland.com. 4. Beijing Research Institute of Chinese Medicine, Beijing University of Chinese Medicine, Beijing, 100029, China. Electronic address: wangcg1119@126.com. 5. School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 102488, China. Electronic address: jing_cao11@163.com. 6. Chenland Nutritionals, Inc, Irvine, CA, 92614, USA. Electronic address: lli@chenland.com. 7. School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 102488, China. Electronic address: 1510010924@qq.com. 8. Chenland Nutritionals, Inc, Irvine, CA, 92614, USA. Electronic address: dliu@chenland.com. 9. Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY, 10065, USA. Electronic address: zhz9010@med.cornell.edu. 10. State Key Laboratory of Proteomics, National Center for Protein Sciences (Beijing), Institute of Lifeomics, Beijing, 102206, China. Electronic address: pharmapapers@hotmail.com. 11. Chenland Nutritionals, Inc, Irvine, CA, 92614, USA; Traditional Chinese Medicine College, Inner Mongolia Medical University, Jinshan Development Zone Hohhot, Inner Mongolia, 010110, China. Electronic address: zhangzengliang1978@outlook.com. 12. School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 102488, China. Electronic address: yshi@bucm.edu.cn.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: Epimedium brevicornu Maxim, Dioscorea nipponica Makino, and Salvia miltiorrhiza Bunge formula (EDS) are three traditional Chinese medicines commonly combined and used to treat osteoarthritis (OA). However, the mechanism of its therapeutic effect on OA is still unclear. AIM OF THE STUDY: The aim of this study was to investigate the potential anti osteoarthritis mechanism of EDS in the treatment of OA rats' model by quantitative proteomics. MATERIALS AND METHODS: A papain-induced rat OA model was established, and then EDS was intragastrically administered for 28 days. A label-free quantification proteomics was performed to evaluate the holistic efficacy of EDS against OA and identify the possible protein profiles mechanisms. The expression levels of critical changed proteins were validated by RT-qPCR and Western blotting. The effects of EDS were then assessed by evaluating pathologic changes in the affected knee joint and measuring pressure pain threshold, acoustic reflex threshold, angle of joint curvature. RESULTS: Proteomics analysis showed that 62 proteins were significantly upregulated and 208 proteins were downregulated in OA group compared to control group. The changed proteins were involved in activation of humoral immunity response, complement cascade activation, leukocyte mediated immunity, acute inflammatory response, endocytosis regulation, and proteolysis regulation. The EDS treatment partially restored the protein profile changes. The protective effects of EDS on pathologic changes in OA rats' knee joint and pain threshold assessment were consisted with the proteomics results. CONCLUSIONS: The results suggest that EDS exerted synergistic therapeutic efficacies to against OA through suppressing inflammation, modulating the immune system, relieving joint pain, and attenuating cartilage degradation.
ETHNOPHARMACOLOGICAL RELEVANCE: Epimedium brevicornu Maxim, Dioscorea nipponica Makino, and Salvia miltiorrhiza Bunge formula (EDS) are three traditional Chinese medicines commonly combined and used to treat osteoarthritis (OA). However, the mechanism of its therapeutic effect on OA is still unclear. AIM OF THE STUDY: The aim of this study was to investigate the potential anti osteoarthritis mechanism of EDS in the treatment of OA rats' model by quantitative proteomics. MATERIALS AND METHODS: A papain-induced rat OA model was established, and then EDS was intragastrically administered for 28 days. A label-free quantification proteomics was performed to evaluate the holistic efficacy of EDS against OA and identify the possible protein profiles mechanisms. The expression levels of critical changed proteins were validated by RT-qPCR and Western blotting. The effects of EDS were then assessed by evaluating pathologic changes in the affected knee joint and measuring pressure pain threshold, acoustic reflex threshold, angle of joint curvature. RESULTS: Proteomics analysis showed that 62 proteins were significantly upregulated and 208 proteins were downregulated in OA group compared to control group. The changed proteins were involved in activation of humoral immunity response, complement cascade activation, leukocyte mediated immunity, acute inflammatory response, endocytosis regulation, and proteolysis regulation. The EDS treatment partially restored the protein profile changes. The protective effects of EDS on pathologic changes in OA rats' knee joint and pain threshold assessment were consisted with the proteomics results. CONCLUSIONS: The results suggest that EDS exerted synergistic therapeutic efficacies to against OA through suppressing inflammation, modulating the immune system, relieving joint pain, and attenuating cartilage degradation.