Page 33 - 《中国药房》2021年20期

Discussion on the Inhibitory Mechanism of Lanthanum Chloride on Vascular Smooth Muscle Cell
Calcification Induced by High Phosphorus Based on NF-κB Signaling Pathway
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GU Chao ，ZHAO Lulu ，LI Gang ，GAO Yuan ，WANG Shengnan ，LI Xiaojia ，YUAN Xiaorong ，WANG
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Qiwen ，Baolechaolu ，HAN Ruilan （1. School of Pharmacy，Inner Mongolia Medical University，Hohhot
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010110，China；2. Collaborative Innovation Center of Mongolian Medicine，Inner Mongolia Medical University，
Hohhot 010110，China）
ABSTRACT OBJECTIVE：To discuss the inhibitory effect of lanthanum chloride on the calcification of vascular smooth muscle
cells（VSMCs）induced by high phosphorus and its mechanism. METHODS：On the basis of screening the action concentration
and time of lanthanum chloride by MTT method，human VSMCs were divided into control group（1 mmol/L phosphorus solution），
lanthanum chloride high concentration control group（1 mmol/L phosphorus solution+60 μmol/L lanthanum chloride），model group
（3 mmol/L phosphorus solution），sodium chloride group（3 mmol/L phosphorus solution+180 μmol/L sodium chloride），nuclear
factor κB（NF-κB）signaling pathway agonist+lanthanum chloride group（3 mmol/L phosphorus solution+1 μg/mL lipopolysaccharide+
60 μmol/L lanthanum chloride），positive control group（3 mmol/L phosphorus solution+100 μmol/L sodium pyrophosphate），and
lanthanum chloride low，medium，and high concentration groups（3 mmol/L phosphorus solution+15，30，60 μmol/L lanthanum
chloride）. Alizarin red S staining and Von Kossa staining were used to detect cell calcification in each group after treated with
phosphorus solution for 6 d and relevant medicine for 2 d. Western blot assay was used to detect the protein expression of TNF-α
receptor associated protein 6（TRAF6），nuclear factor κB inhibitor protein α（IκBα），NF-κB p65，bone morphogenetic protein 2
（BMP-2），smooth muscle 22 α（SM22 α）and Runt related transcription factor 2（Runx2）. Real-time fluorescence quantitative
polymerase chain reaction was used to detect mRNA expression of TRAF6，I κ B α ，BMP-2，SM22 α and Runx2. RESULTS：
Compared with control group，no cell calcification was observed in the lanthanum chloride high concentration control group，while
obvious cell calcification and significant increase of OD value were observed in model group and sodium chloride group（P＜
0.01）；protein and mRNA expression of TRAF6 and BMP-2 in cytoplasm as well as mRNA expression of Runx2，protein
expression of NF-κB p65 and Runx2 in nucleus were significantly increased（P＜0.01）；protein and mRNA expression of IκBα and
SM22α as well as protein expression of NF-κB p65 in cytoplasm were significantly decreased（P＜0.01）. Compared with model
group，cell calcification was significantly improved in lanthanum chloride groups and positive control group，while OD values were
significantly reduced；the expression levels of the above-mentioned protein and mRNA were reversed to varying degrees（P＜0.05
or P＜0.01）. Compared with lanthanum chloride high concentration group，obvious cell calcification was observed in NF-κ B
signaling pathway agonist + lanthanum chloride group， and OD value was significantly increased； the above indexes were
significantly reversed in cytoplasm and nucleus （P＜0.05 or P＜0.01）. CONCLUSIONS：Lanthanum chloride can inhibit the
calcification of VSMCs induced by high phosphorus，and its mechanism may be related to the inhibition of NF-κ B signaling
pathway activation.
KEYWORDS Lanthanum chloride；High phosphorus；Vascular calcification；NF-κB signaling pathway；Human vascular smooth
muscle cells

全球疾病负担报告显示，2017年慢性肾脏病（CKD） 细胞转分化，提示这一信号通路可作为抑制血管中层钙
的患病人数约为 6.975 亿，其中 120 万人死于肾衰竭，另 化的潜在靶标 [5－7] 。
有 140 万人死于并发的心血管疾病 。已有研究表明， 本课题组前期已经证明，氢氧化镧可改善 CKD 高
[1]
[8]
随着肾功能损伤进展到晚期、功能性肾单位量减少、磷 磷血症模型大鼠的肾功能，降低其血清磷含量 。除此
酸盐排泄受阻，再加上持续摄入饮食，CKD 患者血清磷 之外，还发现纳米氢氧化镧与碳酸镧相比具有更高的效
酸盐浓度将持续升高，最终可能引起血管钙化等并发症 价 。由于氢氧化镧难溶于水，不适宜用于细胞实验，因
[9]
[2]
的发生，从而导致CKD患者过早死亡 。 此本研究以氯化镧作为研究对象，基于NF-κB信号通路
血管中层钙化是导致CKD患者心血管疾病死亡的 探究其对高磷致VSMCs 钙化的影响，旨在为CKD并发
[3]
关键因素 。在高磷条件下，血管平滑肌细胞（VSMCs） 症——血管钙化的治疗提供参考。
能够向成骨样细胞转分化，转分化后的 VSMCs 可促进 1 材料
钙和磷酸盐沉淀以及磷酸钙晶体生长，进而促进动脉中 1.1 主要仪器
层钙化 。有研究表明，细胞外聚积的磷酸盐能通过激 本研究所用主要仪器包括 PikoReal 96 型荧光定量
[4]
活核因子κB（NF-κB）信号通路而诱导VSMCs向成骨样 聚合酶链反应（PCR）仪、Heraeus HERAcell 150i 型 CO2

中国药房 2021年第32卷第20期 China Pharmacy 2021 Vol. 32 No. 20 ·2459 ·