【摘要】：背景:晚期糖基化終末產(chǎn)物對破骨細(xì)胞骨吸收功能的影響存在爭議,作者的前期研究表明晚期糖基化終末產(chǎn)物作用于破骨細(xì)胞前體可顯著抑制骨吸收功能,但關(guān)于其對破骨細(xì)胞泌酸功能的影響尚不明晰。目的:探究晚期糖基化終末產(chǎn)物對破骨細(xì)胞泌酸功能的影響及其機(jī)制。方法:以15μg/L的RANKL對破骨細(xì)胞前體RAW 264.7進(jìn)行定向誘導(dǎo)(正常組),實驗組中另加入50-400 mg/L不等的晚期糖基化終末產(chǎn)物及對照用牛血清白蛋白(100 mg/L)。通過骨吸收實驗驗證晚期糖基化終末產(chǎn)物對骨吸收的抑制效應(yīng),并以吖啶橙染色觀察晚期糖基化終末產(chǎn)物對破骨細(xì)胞泌酸功能的影響;進(jìn)一步檢測質(zhì)子泵V-ATPase a3及氯離子通道ClC-7的表達(dá)情況,分析晚期糖基化終末產(chǎn)物影響破骨細(xì)胞泌酸的相關(guān)機(jī)制。結(jié)果與結(jié)論:(1)晚期糖基化終末產(chǎn)物組的骨吸收面積較正常組顯著減少(P0.05);(2)吖啶橙染色顯示晚期糖基化終末產(chǎn)物組的紅色熒光(620 nm)強(qiáng)度較正常組顯著減少(P0.05),抑制程度隨著刺激濃度的增高而加重;(3)免疫細(xì)胞化學(xué)染色、蛋白質(zhì)免疫印跡及PCR發(fā)現(xiàn),晚期糖基化終末產(chǎn)物組的V-ATPase a3及ClC-7表達(dá)量均較正常組顯著下降(P0.05);(4)綜上結(jié)果表明,晚期糖基化終末產(chǎn)物作用于破骨細(xì)胞前體可顯著抑制破骨細(xì)胞的泌酸功能,其機(jī)制可能與晚期糖基化終末產(chǎn)物抑制了質(zhì)子泵V-ATPase a3及氯離子通道ClC-7的表達(dá)相關(guān)。
[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.
【作者單位】： 中山大學(xué)附屬第一醫(yī)院關(guān)節(jié)外科;中山大學(xué)中山醫(yī)學(xué)院組織胚胎學(xué)教研室;
【基金】：國家自然科學(xué)基金面上項目(81672149),資助單位:中山大學(xué)附屬第一醫(yī)院 廣東省自然科學(xué)基金-重點(2015A030311004),資助單位:中山大學(xué)附屬第一醫(yī)院~~
【分類號】：R68
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本文編號：2262293