| 张婷婷,李雪萍.低强度脉冲超声和吡格列酮对脂多糖诱导的OA软骨细胞IGF-1/mTOR/PGE2通路的影响[J].中国康复,2018,33(3):179-183 |
| 低强度脉冲超声和吡格列酮对脂多糖诱导的OA软骨细胞IGF-1/mTOR/PGE2通路的影响 |
| Effect of LIPUS and pioglitazone on IGF-1/mTOR/PGE2 pathway in LPS-induced OA chondrocytes |
| |
| DOI： |
| 中文关键词: 低强度脉冲超声 吡格列酮 骨关节炎 脂多糖 |
| 英文关键词: Low intensity pulsed ultrasound Pioglitazone Osteoarthritis Lipopolysaccharide |
| 基金项目:国家自然科学基金(81772437) |
|
| 摘要点击次数: 6937 |
| 全文下载次数: 4604 |
| 中文摘要: |
| 目的：探讨低强度脉冲超声(LIPUS)和吡格列酮对脂多糖(LPS）诱导的骨关节炎(OA)软骨细胞经雷帕霉素靶蛋白(mTOR)途径的软骨保护作用比较。方法：于新西兰大白兔膝关节软骨提取正常软骨细胞，分别进行体外培养并按数字表法随机分为4组:正常组、LPS组、LIPUS组（LPS+LIPUS）、吡格列酮组（LPS+吡格列酮,Pioglitazone组）各6只。LIPUS干预选取40 mW/cm2强度，每次照射20min，1次/d，共7d。Pioglitazone组的软骨细胞在含有100uM吡格列酮培养基中连续培养7d。于第7天应用CCK-8检测各组软骨细胞增殖活性，免疫细胞化学染色法检测各组软骨细胞的II型胶原(COL2)的表达情况，ELISA技术检测胰岛素生长因子1(IGF-1)、前列腺素E2(PGE2）的表达水平，RT-PCR和Western blot技术检测mTORmRNA表达水平和蛋白含量。结果：①免疫细胞化学染色：正常组软骨细胞内的COL2表达较LPS组高（P＜0.05）。②CCK-8：100uM吡格列酮对软骨细胞的增殖作用较1uM,10uM,50uM吡格列酮强（P＜0.05）。③ELISA：与LPS组相比,Pioglitazone组和LIPUS组的软骨细胞IGF-1表达水平均上升（P＜0.05），并且LIPUS组较Pioglitazone组上升明显（P＜0.05）。与LPS组相比，LIPUS组和Pioglitazone组的软骨细胞PGE2表达水平明显下降（P＜0.05）；且Pioglitazone组PGE2水平较LIPUS组下降明显（P＜0.05）。④RT-PCR和Western blot：与LPS组相比,LIPUS组和Pioglitazone组的软骨细胞mTOR mRNA水平和蛋白含量明显下降（P＜0.05），且Pioglitazone组mTOR mRNA水平和蛋白含量较LIPUS组明显下降（P＜0.05）。结论：LIPUS与吡格列酮可能通过IGF-1/mTOR/PGE2信号通路上调软骨细胞IGF-1的水平，下调PGE2和mTOR的表达，加强软骨细胞自我保护作用。 |
| 英文摘要: |
| 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. |
|
查看全文
下载PDF阅读器 HTML全文 |
| 关闭 |
|
|
|
