慢性氟中毒大鼠骨组织Gli1表达变化及其与β-catenin的相关性分析
首发时间:2018-09-07
摘要:目的观察使用刺猬蛋白(Hedgehog, Hh)信号阻断剂环巴胺(Cyclopamine, Cycl)对慢性氟中毒大鼠骨组织中神经胶质瘤相关癌基因同源蛋白1(Glioma-associated oncogene homolog 1, Gli1)及β链蛋白(β-catenin)表达的影响,探讨两者在氟致骨损伤中的意义。方法 SD大鼠48只随机分为4组,每组12只。对照组饮用自来水(含氟量<1 mg/L),氟中毒组(NaF50 mg/L)、氟+阻断剂组(氟+Cycl组)和氟+阻断剂对照组(氟+DMSO组)分别饮用含50 mg/L氟化钠的自来水。饲养6个月后分别给氟+ Cycl组和氟+DMSO组大鼠腹腔注射Cycl和DMSO。检测各组大鼠尿氟含量;酶联免疫吸附试验测定血清骨碱性磷酸酶(BALP)含量;实时荧光定量PCR法、免疫组织化学和蛋白印迹法检测骨组织中Gli1和β-catenin的mRNA和蛋白表达情况。结果染氟后大鼠尿氟含量明显增加,氟中毒组、氟+Cycl组和氟+DMSO组较对照组明显升高,但三组间未见明显差异。氟中毒组较对照组血清BALP含量增加,Cycl阻断后血清BALP含量明显降低(P<0.05)。与对照组比较,氟中毒组Gli1和β-catenin mRNA和蛋白均有表达增强。使用Cycl后,两者表达均较氟中毒组降低(P<0.05),而阻断对照组则无明显变化。Gli1与β-catenin的表达呈正相关关系(r=0.476,P<0.05)。结论氟致Hh信号表达增强可能促进β-catenin的表达,并且两者具有协同作用,而这在一定程度上增加了骨形成。
关键词: 氟中毒 骨 神经胶质瘤相关癌基因-1 β链蛋白
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Relationship between expression of Gli1 and β-catenin on rats\' bone with chronic fluorosis
Abstract:To explore the expression and significance of Glioma-associated oncogene homolog 1 (Gli1) and β-catenin on bone of rats with chronic fluorosis which were inhibited by Cyclopamine (Cycl). Methods Forty-eight healthy SD rats were devided to four groups with the method of random digits table. The rats of control were fed with tap water (NaF<1 mg/L) and the experimental rats were exposed to NaF (50 mg/L) added to the drinking water to established the chronic fluorosis model. Then Cycl or DMSO were administered by intraperitoneal injection in rats of F+Cycl or F+DMSO group respectively. Bone fluorine was detected by ashing-fluorin ion selective electrode method. The content of bone alkaline phosphatase (BALP) in rats’ serum was detected by enzyme-linked immunosorbent assay (ELISA). The expression of Gli1 and β-catenin mRNA and protein on bone were detected by Real-time PCR, immunohistochemistry (IHC) and Western Blot. Results The contents of urine fluoride was increased in the experimental groups compared with the control, but there is no significance. The content of BALP was increased in the experimental group and decreased in F+Cycl group (P<0.05) . Compared with the control, the expression of Gli1 and β-catenin mRNA and protein were increased. Both of them were decreased than the rats in F group. There was positive correlation between the expression of Gli1 and β-catenin (r=0.476, P<0.05). Conlusion It may enhance the expression of β-catenin induced by fluorosis via the Hh sinaling. And they can interact with each other. It may be involved in the osteogenesis to some extent.
Keywords: Fluorosis Bone Glioma-associated oncogene homolog 1 β-catenin
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慢性氟中毒大鼠骨组织Gli1表达变化及其与β-catenin的相关性分析
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