| Effect of LIPUS and pioglitazone on IGF-1/mTOR/PGE2 pathway in LPS-induced OA chondrocytes |
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| DOI： |
| EN KeyWords: Low intensity pulsed ultrasound Pioglitazone Osteoarthritis Lipopolysaccharide |
| Fund Project:国家自然科学基金(81772437) |
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| EN Abstract: |
| Objective: To investigate the protective effect of low intensity pulsed ultrasound (LIPUS) and pioglitazone on lipopolysaccharide (LPS)-induced OA chondrocytes through mammalian target of rapamycin (mTOR) signal. Methods: The normal chondrocytes were extracted from the knee of New Zealand rabbits and randomly divided into four groups: normal group, LPS group, LIPUS group (LPS+LIPUS), pioglitazone group (LPS+pioglitazone). The LIPUS group received LIPUS intervention with intensity of 40 mW/cm2, 20 min every day for 7 days. The chondrocytes in pioglitazone group were incubated with pioglitazone for 7 days. On the 7th day, the proliferation of chondrocytes was measured by CCK-8 assay. The expression of type II collagen (COL2) in chondrocytes was detected by immunocytochemistry. The levels of insulin-like growth factor-1 (IGF-1) and prostaglandin (PG) E2 in the supernatants of chondrocytes were quantified by ELISA. The mTOR mRNA and protein expression was detected by RT-PCR and Western blot respectively. Results: ①Immunocytochemical staining: The expression of COL2 in chondrocytes of normal group was higher than LPS group. ②CCK-8: The proliferation of chondrocytes in 100 μM pioglitazone group was stronger than 1 μM, 10 μM, and 50 μM pioglitazone groups. ③ELISA: Compared with LPS group, the expression of IGF-1 in pioglitazone group and LIPUS group increased with the difference being statistically significant (P<0.05), and the LIPUS group had a significant increase compared with the pioglitazone group. Compared with LPS group, the expression of PGE2 in LIPUS group and pioglitazone group was significantly decreased (P<0.05), and PGE2 level in pioglitazone group was significantly lower than that in LIPUS group. ④RT-PCR and Western blot: Compared with LPS group, mTOR mRNA and protein expression in LIPUS group and pioglitazone group was significantly decreased (P<0.05), and the expression was significantly lower than that of the LIPUS group. Conclusion: Both LIPUS and pioglitazone can increase the level of IGF-1 of cartilage cells and decrease the expression of PEG2 and mTOR through the IGF-1/mTOR/PGE2 signaling pathway, enhance the self-protection of chondrocytes. |
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