晚期糖基化终末产物可通过调节V-ATPase a3与ClC-7影响破骨细胞的泌酸功能
发布时间：2018-10-10 15:27
【摘要】：背景:晚期糖基化终末产物对破骨细胞骨吸收功能的影响存在争议,作者的前期研究表明晚期糖基化终末产物作用于破骨细胞前体可显著抑制骨吸收功能,但关于其对破骨细胞泌酸功能的影响尚不明晰。目的:探究晚期糖基化终末产物对破骨细胞泌酸功能的影响及其机制。方法:以15μg/L的RANKL对破骨细胞前体RAW 264.7进行定向诱导(正常组),实验组中另加入50-400 mg/L不等的晚期糖基化终末产物及对照用牛血清白蛋白(100 mg/L)。通过骨吸收实验验证晚期糖基化终末产物对骨吸收的抑制效应,并以吖啶橙染色观察晚期糖基化终末产物对破骨细胞泌酸功能的影响;进一步检测质子泵V-ATPase a3及氯离子通道ClC-7的表达情况,分析晚期糖基化终末产物影响破骨细胞泌酸的相关机制。结果与结论:(1)晚期糖基化终末产物组的骨吸收面积较正常组显著减少(P0.05);(2)吖啶橙染色显示晚期糖基化终末产物组的红色荧光(620 nm)强度较正常组显著减少(P0.05),抑制程度随着刺激浓度的增高而加重;(3)免疫细胞化学染色、蛋白质免疫印迹及PCR发现,晚期糖基化终末产物组的V-ATPase a3及ClC-7表达量均较正常组显著下降(P0.05);(4)综上结果表明,晚期糖基化终末产物作用于破骨细胞前体可显著抑制破骨细胞的泌酸功能,其机制可能与晚期糖基化终末产物抑制了质子泵V-ATPase a3及氯离子通道ClC-7的表达相关。
[Abstract]:Background: the effects of advanced glycation end products on osteoclastic bone resorption are controversial. Previous studies by the authors have shown that late glycosylation end products can significantly inhibit bone resorption by acting on osteoclast precursors. However, its effect on the acid secretion of osteoclasts remains unclear. Aim: to investigate the effect of advanced glycation end products on acid secretion of osteoclasts and its mechanism. Methods: the osteoclast precursor RAW 264.7 was induced by 15 渭 g / L RANKL (normal group). The late glycosylation end product (50 to 400 mg/L) and bovine serum albumin (100 mg/L) were added in the experimental group. The inhibitory effect of advanced glycation end products on bone resorption was verified by bone resorption test, and the effect of acridine orange staining on the acid secretion of osteoclasts was observed by acridine orange staining. Furthermore, the expression of proton pump V-ATPase a3 and chloride channel ClC-7 was detected, and the mechanism of late glycation end products affecting the acid secretion of osteoclasts was analyzed. Results and conclusion: (1) the bone resorption area in the advanced glycosylation end product group was significantly lower than that in the normal group (P0.05); (2) acridine orange staining showed that the red fluorescence (620 nm) intensity in the advanced glycosylation end product group was significantly lower than that in the normal group (P0.05), and the inhibition degree was significant (P0.05). (3) immunocytochemical staining, Western blot and PCR showed that the expression of V-ATPase a3 and ClC-7 in the advanced glycosylation end product group was significantly lower than that in the normal group (P0.05); (4). The late glycation end products could inhibit the acid secretion of osteoclasts significantly by acting on the precursor of osteoclasts. The mechanism may be related to the inhibition of proton pump V-ATPase a3 and chloride channel ClC-7 expression by the late glycation end products.
【作者单位】： 中山大学附属第一医院关节外科;中山大学中山医学院组织胚胎学教研室;
【基金】：国家自然科学基金面上项目(81672149),资助单位:中山大学附属第一医院 广东省自然科学基金-重点(2015A030311004),资助单位:中山大学附属第一医院~~
【分类号】：R68
，
本文编号：2262293
[Abstract]:Background: the effects of advanced glycation end products on osteoclastic bone resorption are controversial. Previous studies by the authors have shown that late glycosylation end products can significantly inhibit bone resorption by acting on osteoclast precursors. However, its effect on the acid secretion of osteoclasts remains unclear. Aim: to investigate the effect of advanced glycation end products on acid secretion of osteoclasts and its mechanism. Methods: the osteoclast precursor RAW 264.7 was induced by 15 渭 g / L RANKL (normal group). The late glycosylation end product (50 to 400 mg/L) and bovine serum albumin (100 mg/L) were added in the experimental group. The inhibitory effect of advanced glycation end products on bone resorption was verified by bone resorption test, and the effect of acridine orange staining on the acid secretion of osteoclasts was observed by acridine orange staining. Furthermore, the expression of proton pump V-ATPase a3 and chloride channel ClC-7 was detected, and the mechanism of late glycation end products affecting the acid secretion of osteoclasts was analyzed. Results and conclusion: (1) the bone resorption area in the advanced glycosylation end product group was significantly lower than that in the normal group (P0.05); (2) acridine orange staining showed that the red fluorescence (620 nm) intensity in the advanced glycosylation end product group was significantly lower than that in the normal group (P0.05), and the inhibition degree was significant (P0.05). (3) immunocytochemical staining, Western blot and PCR showed that the expression of V-ATPase a3 and ClC-7 in the advanced glycosylation end product group was significantly lower than that in the normal group (P0.05); (4). The late glycation end products could inhibit the acid secretion of osteoclasts significantly by acting on the precursor of osteoclasts. The mechanism may be related to the inhibition of proton pump V-ATPase a3 and chloride channel ClC-7 expression by the late glycation end products.
【作者单位】： 中山大学附属第一医院关节外科;中山大学中山医学院组织胚胎学教研室;
【基金】：国家自然科学基金面上项目(81672149),资助单位:中山大学附属第一医院 广东省自然科学基金-重点(2015A030311004),资助单位:中山大学附属第一医院~~
【分类号】：R68
，
本文编号：2262293
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