Hh信号通路在氟致大鼠原代软骨细胞损伤中作用

发布时间：2018-09-08 12:42

【摘要】：目的探讨体外染氟软骨细胞中刺猥蛋白基因(Hh)信号通路中音速刺猬蛋白、跨膜蛋白(Smo)及下游骨形态发生蛋白-2(BMP-2)因子表达在氟中毒软骨细胞损伤中的作用。方法取1周龄健康SD大鼠,用机械、酶消化法分离膝关节软骨细胞,免疫细胞化学对原代软骨细胞进行鉴定。实验设对照组和染氟组(5、10、20、40 mg/L)。培养48 h后,噻唑蓝法检测各组细胞活力,蛋白印迹及逆转录PCR分别检测各组细胞Smo、Shh、BMP-2蛋白及mRNA表达;流式细胞术检测各组细胞凋亡情况。结果与对照组比较,5、10 mg/L染氟组细胞活力[分别为(113.33±11.74)%、(127.25±10.24)%]明显升高,40 mg/L染氟组细胞活力[(73.91±9.94)%]明显下降(P0.05);染氟组细胞中Shh、Smo、BMP-2蛋白和mRNA的表达量随染氟浓度增加而升高;40 mg/L染氟组细胞凋亡率为(11.13±1.20)%,明显高于对照组。结论氟可激活软骨细胞中Hh信号通路,Hh信号通路与软骨细胞凋亡可能共同参与氟中毒软骨损害过程。
[Abstract]:Objective to investigate the role of sonic hedgehog protein, transmembrane protein (Smo) and downstream bone morphogenetic protein (BMP-2) factor expression in the (Hh) signaling pathway of impure protein gene in fluorosis chondrocytes in vitro. Methods the chondrocytes of knee joint were isolated by mechanical and enzymatic digestion from 1 week old healthy SD rats. The primary chondrocytes were identified by immunocytochemistry. The experiment was divided into two groups: control group and fluorine-exposed group (5 ~ 10 ~ 20 ~ 40 mg/L). After cultured for 48 h, the cell viability was detected by thiazolyl blue assay, the expression of Smo,Shh,BMP-2 protein and mRNA were detected by Western blot and reverse transcription PCR, and the apoptosis of each group was detected by flow cytometry. Results compared with the control group, the cell viability [(113.33 卤11.74), (127.25 卤10.24)%] of the 10 mg/L fluorine group increased significantly [(73.91 卤9.94)%] significantly higher than that of the control group (P0.05), and the expression of Shh,Smo,BMP-2 protein and mRNA increased with the increase of fluoride concentration in the fluorine-exposed group. The apoptotic rate was (11.13 卤1.20) in the control group, which was significantly higher than that in the control group. Conclusion fluoride can activate the Hh signaling pathway in chondrocytes and the apoptosis of chondrocytes may be involved in the process of cartilage damage caused by fluorosis.
【作者单位】：贵州医科大学病理学教研室;
【基金】：国家自然科学基金(81260419) 教育部博士点基金(博导类)(20125215110001)
【分类号】：R114

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