摘要
目的 观察低强度脉冲超声（LIPUS）和吡格列酮（pioglitazone）经过氧化物酶增殖物激活受体γ（PPARγ）/核转录因子-κB（NF-κB）/诱导型一氧化氮合酶（iNOS）途径对骨关节炎（OA）软骨细胞的保护作用。 方法 选取健康成年新西兰大白兔24只，按随机数字表法分为正常组、脂多糖（LPS）组、LIPUS组（LPS＋LIPUS）、吡格列酮组（LPS＋吡格列酮），每组6只大白兔。分别提取4组大白兔膝正常软骨细胞,采用ELISA法检测4组软骨细胞肿瘤坏死因子-α（TNF-α）、瘦素（LEP）、一氧化氮（NO）的水平；采用免疫细胞化学和免疫荧光染色法检测4组软骨细胞Ⅱ型胶原（COL2）的表达情况；另采用实时荧光定量逆转录-聚合酶链反应（RT-PCR）和免疫蛋白印迹法检测（Western blot）技术分别检测4组软骨细胞的PPARγ，NF-κB，iNOS mRNA和蛋白表达水平。 结果 LIPUS组和吡格列酮组OA软骨细胞的TNF-α、LEP、NO水平与LPS组比较，均显著下降，差异均有统计学意义（P〈0.05），且吡格列酮组下调幅度大于LIPUS组，差异有统计学意义（P〈0.05）；LIPUS组OA软骨细胞COL2的表达水平显著上调，与LPS组比较，差异有统计学意义（P〈0.05）；LIPUS组和吡格列酮组OA软骨细胞PPARγ的mRNA和蛋白表达水平均高于LPS组，差异均有统计学意义（P〈0.05）；吡格列酮组和LIPUS组OA软骨细胞NF-κB、iNOS mRNA和蛋白表达下降，与LPS组比较，差异均有统计学意义（P〈0.05），且吡格列酮组下降程度大于LIPUS组，差异有统计学意义（P〈0.05）。 结论 LIPUS和吡格列酮均可能通过PPARγ/NF-κB/iNOS途径参与OA软骨细胞的抗炎症、COL2合成过程，起到OA软骨细胞的保护作用。
Objective To investigate any protective effect of low-intensity pulsed ultrasound （LIPUS） and pioglitazone on chondrocytes in osteoarthritic patients using the pathway from peroxisome proliferator-activated γreceptor （PPARγ） to nuclear factor kappa B （NF-κB） to inducible nitric oxide synthase （iNOS）. Methods Normal chondrocytes of 24 healthy adult New Zealand white rabbits were extracted and divided into a normal group, a lipopolysaccharide （LPS） group, a LIPUS group （LPS＋LIPUS） and a pioglitazone group （LPS＋pioglitazone）, each of 6 using a random number table. Each group was given the intervention their names implies. The levels of tumor necrosis factor-α （TNF-α）, leptin （LEP） and nitric oxide （NO） in the chondrocytes were detected using enzyme-linked immune sorbent assays. The expression of type II collagen （COL2） in the chondrocytes of each groups was detected using immunocytochemistry and fluorescent staining. The mRNA and protein expressions of PPARγ, NF-κB and iNOS were detected using reverse transcription polymerase chain reactions and western blotting respectively. Results Compared with the LPS group, the average level of TNF-α, LEP and NO in the LIPUS and pioglitazone groups was significantly lower, with the levels in the pioglitazone group significantly lower than in the LIPUS group. Compared with the LPS group, COL2 expression in the LIPUS group was significantly greater. The mRNA and protein expressions of PPARγ in the chondrocytes in the LIPUS and pioglitazone groups were significantly higher than those in the LPS group. Compared with the LPS group, the mRNA and protein expressions of NF-κB and iNOS in the pioglitazone and LIPUS groups were significantly lower, with the pioglitazone group′s levels significantly below those of the LIPUS group. Conclusion LIPUS and pioglitazone may promote anti-inflammatory action and COL2 synthesis in chondrocytes through the PPARγ/ NF-κB/iNOS pathway and play a protective role, at least in rabbits.
作者
张婷婷
高明霞
夏鹏
任莎莎
王馨伟
程凯
林强
李雪萍
Zhang Tingting , Gao Mingxia, Xia Peng, Ren Shasha, Wang Xinwei, Cheng Kai, Lin Qiang, Li Xueping(Department of Rehabilitation Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, Chin)
出处
《中华物理医学与康复杂志》
CAS
CSCD
北大核心
2018年第3期167-173,共7页
Chinese Journal of Physical Medicine and Rehabilitation
基金
国家自然科学基金（81772437）
国家自然科学基金（81501941）
