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毫米波治疗通过 p38MAPK 通路抑制一氧化氮诱导的软骨细胞凋亡。

Millimeter wave treatment inhibits NO-induced apoptosis of chondrocytes through the p38MAPK pathway.

机构信息

Huatuo, University Town, Minhou Shangjie, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350108, P.R. China.

出版信息

Int J Mol Med. 2010 Mar;25(3):393-9. doi: 10.3892/ijmm_00000357.

Abstract

In the present study, we investigated the effects of millimeter wave treatment on the activation of the p38MAPK signaling pathway in the process of NO-induced apoptosis in chondrocytes. Cartilage was isolated from the knee joint of SD rats and used to establish cultured primary chondrocytes. After identification using in situ staining of type II collagen, the passage 2 chondrocytes were incubated with or without sodium nitroprussiate (SNP) to induce apoptosis and treated with a millimeter wave for various times. The apoptosis of chondrocytes was detected using immunofluorescence, an MTT assay, and Annexin V-FITC labeling followed by fluorescence-activated cell sorting (FACS). The activity of caspase-3 was measured using colorimeters, and the levels of p38 and p53 were also detected using RT-PCR and Western blotting. After treatment with SNP, the OD values of the experimental groups were significantly lower than the control group (P<0.01). The 24-h interference of a millimeter wave significantly prevented apoptosis (P<0.01) and showed a dose dependency, and an identical trend of apoptosis was noted with normal cell number counting (P<0.01) and FACS (P<0.01). Consistently, the caspase 3 activity showed a reverse trend, with the highest activity in the experimental group receiving no millimeter wave treatment (P<0.01). The mRNA expression of p38 and p53 and the protein levels of phosphorylated p38 and p53 showed a similar trend (P<0.01) to that of caspase 3 activity. In conclusion, millimeter wave treatment inhibits the SNP-induced apoptosis of chondrocytes through the p38MAPK pathway.

摘要

在本研究中,我们研究了毫米波治疗对 NO 诱导软骨细胞凋亡过程中 p38MAPK 信号通路激活的影响。从小鼠膝关节分离软骨,用于建立原代培养的软骨细胞。经 II 型胶原原位染色鉴定后,将第 2 代软骨细胞与硝普钠(SNP)孵育或不孵育以诱导凋亡,并进行不同时间的毫米波治疗。采用免疫荧光、MTT 检测和 Annexin V-FITC 标记后荧光激活细胞分选(FACS)检测软骨细胞凋亡。采用比色法测定 caspase-3 的活性,采用 RT-PCR 和 Western blot 检测 p38 和 p53 的水平。SNP 处理后,实验组的 OD 值明显低于对照组(P<0.01)。24 小时毫米波干预明显预防了凋亡(P<0.01),并表现出剂量依赖性,与正常细胞计数(P<0.01)和 FACS(P<0.01)的凋亡趋势相同。同样,caspase 3 的活性呈现相反的趋势,未接受毫米波治疗的实验组活性最高(P<0.01)。p38 和 p53 的 mRNA 表达以及磷酸化 p38 和 p53 的蛋白水平与 caspase 3 活性表现出相似的趋势(P<0.01)。总之,毫米波治疗通过 p38MAPK 通路抑制 SNP 诱导的软骨细胞凋亡。

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