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[严重烧伤大鼠骨形成与骨吸收相关指标的变化]

[Changes in the related indicators of bone formation and bone resorption in severely burned rats].

作者信息

Gong X, Ye Z Q, Yu G, Zhang W, Zhang W D, Zhou X Q, Li M, Xie W G

机构信息

Institute of Burns, Tongren Hospital of Wuhan University & Wuhan Third Hospital, Wuhan 430060, China.

出版信息

Zhonghua Shao Shang Za Zhi. 2021 Sep 20;37(9):839-845. doi: 10.3760/cma.j.cn501120-20200505-00253.

Abstract

To observe the changes in the related indicators of bone formation and bone resorption in severely burned rats. The experimental research method was adopted. Thirty female Sprague-Dawley rats aged 6 to 8 weeks were divided into sham injury group, 12% total body surface area (TBSA) full-thickness burn group, and 24%TBSA full-thickness burn group according to the random number table, with 10 rats in each group. The rats were treated on the back correspondingly, after which, the burned rats were rehydrated by intraperitoneal injection according to the Parkland formula, and the wound was coated with 20 g/L iodophor until wound healing. On post injury day (PID) 28, the tibia tissue of rats in each group was collected. The new bone tissue and the number of osteoclasts were observed after staining with Masson and tartrate-resistant acid phosphatase, respectively. The abdominal aortic blood of rats in each group was harvested for serum preparation. The bone metabolism indexes of serum calcium ion and phosphorus ion concentration were determined by the methyl thymol blue colorimetric method and phosphomolybdic acid method, respectively. The serum levels of bone formation marker of aminoterminal propeptide of type 1 procollagen (P1NP) and bone resorption marker of beta-carboxy-terminated peptide of type Ⅰ collagen (β-CTX) were determined by enzyme-linked immunosorbent assay. The first lumbar spine tissue of rats in each group was collected, and the mRNA expression levels of osteoprotegerin, receptor activator of nuclear factor-κB ligand (RANKL), tumor necrosis factor receptor-associated factor 6 (TRAF-6), nuclear factor of activated T cell 1 (NFATC1), c-Fos, and c-Src were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were statistically analyzed with one-way analysis of variance, Bonferroni method, Welch test, Games-Howell test, Kruskal-Wallis test, Mann-Whitney test, and Bonferroni correction. On PID 28, compared with that in sham injury group, the formation of new bone tissue in the tibia tissue of rats in the two burn groups was decreased, and the larger the burn area, the more obvious the decrease. The numbers of osteoclasts in the tibia tissue of rats in the two burn groups were similar, both significantly more than the number in sham injury group. On PID 28, the serum calcium ion concentration and serum level of β-CTX of rats in the three groups were similar (>0.05). The serum phosphorus ion concentration of rats in 24%TBSA full-thickness burn group was significantly higher than that in 12%TBSA full-thickness burn group (<0.05), and the serum phosphorus ion concentrations in the two burn groups were significantly higher than the concentration in sham injury group (<0.01). The serum level of P1NP of rats in 24%TBSA full-thickness burn group was significantly lower than that in sham injury group (<0.01). On PID 28, the mRNA expression levels of osteoprotegerin in the first lumbar spine tissue of rats in sham injury group, 12%TBSA full-thickness burn group, and 24%TBSA full-thickness burn group were 1.01±0.20, 1.71±0.83, and 2.24±0.51, respectively, and that in 24%TBSA full-thickness burn group was significantly higher than that in sham injury group (<0.01). The mRNA expression level of RANKL in the first lumbar spine tissue of rats in 24%TBSA full-thickness burn group was 1.31±0.17, which was significantly higher than 1.00±0.14 in sham injury group and 0.97±0.10 in 12%TBSA full-thickness burn group (<0.01). The mRNA expression levels of TRAF-6, NFATC1 (=3.141, 3.782), and c-Src in the first lumbar tissue of rats in 12%TBSA full-thickness burn group and 24%TBSA full-thickness burn group and the mRNA expression level of c-Fos in the first lumbar tissue of rats in 12%TBSA full-thickness burn group were significantly higher than those in sham injury group (<0.05 or <0.01). The mRNA expression levels of c-Fos and c-Src in the first lumbar spine tissue of rats in 12%TBSA full-thickness burn group were significantly higher than those in 24%TBSA full-thickness burn group (<0.01). Severe burns can cause a decrease in the generation of new bone tissue, an increase in the number of osteoclasts and the serum phosphorus ion concentration, and a decrease in the serum level of P1NP in rats. The level of osteoprotegerin, RANKL, TRAF-6, NFATC1, c-Fos, and c-Src in bone tissue showed an increasing trend while the level of NFATC1, c-Fos, and c-Src showed a decreasing trend with the increase of burn area.

摘要

观察重度烧伤大鼠骨形成和骨吸收相关指标的变化。采用实验研究方法。将30只6至8周龄的雌性Sprague-Dawley大鼠按随机数字表分为假伤组、12%总体表面积(TBSA)全层烧伤组和24%TBSA全层烧伤组,每组10只。对大鼠背部进行相应处理,之后,根据Parkland公式对烧伤大鼠进行腹腔注射补液,并在伤口涂抹20 g/L碘伏直至伤口愈合。在伤后第28天(PID 28),收集每组大鼠的胫骨组织。分别用Masson染色和抗酒石酸酸性磷酸酶染色观察新骨组织和破骨细胞数量。采集每组大鼠的腹主动脉血制备血清。分别用甲基麝香草酚蓝比色法和磷钼酸法测定血清钙离子和磷离子浓度的骨代谢指标。用酶联免疫吸附测定法测定血清Ⅰ型前胶原氨基端前肽(P1NP)的骨形成标志物水平和Ⅰ型胶原β羧基端肽(β-CTX)的骨吸收标志物水平。收集每组大鼠的第一腰椎组织,采用实时荧光定量逆转录聚合酶链反应检测骨保护素(osteoprotegerin)、核因子κB受体激活剂配体(RANKL)、肿瘤坏死因子受体相关因子6(TRAF-6)、活化T细胞核因子1(NFATC1)、c-Fos和c-Src的mRNA表达水平。数据采用单因素方差分析、Bonferroni法、Welch检验、Games-Howell检验、Kruskal-Wallis检验、Mann-Whitney检验和Bonferroni校正进行统计学分析。在PID 28时,与假伤组相比,两个烧伤组大鼠胫骨组织中新骨组织形成减少,且烧伤面积越大,减少越明显。两个烧伤组大鼠胫骨组织中的破骨细胞数量相似,均显著多于假伤组。在PID 28时,三组大鼠的血清钙离子浓度和β-CTX血清水平相似(>0.05)。24%TBSA全层烧伤组大鼠的血清磷离子浓度显著高于12%TBSA全层烧伤组(<0.05),且两个烧伤组的血清磷离子浓度均显著高于假伤组(<0.01)。24%TBSA全层烧伤组大鼠的血清P1NP水平显著低于假伤组(<0.01)。在PID 28时,假伤组、12%TBSA全层烧伤组和24%TBSA全层烧伤组大鼠第一腰椎组织中骨保护素的mRNA表达水平分别为1.01±0.20、1.71±0.83和2.24±0.51,24%TBSA全层烧伤组显著高于假伤组(<0.01)。24%TBSA全层烧伤组大鼠第一腰椎组织中RANKL的mRNA表达水平为1.31±0.17,显著高于假伤组的1.00±0.14和12%TBSA全层烧伤组的0.97±0.10(<0.01)。12%TBSA全层烧伤组和24%TBSA全层烧伤组大鼠第一腰椎组织中TRAF-6、NFATC1(=3.141,3.782)和c-Src的mRNA表达水平以及12%TBSA全层烧伤组大鼠第一腰椎组织中c-Fos的mRNA表达水平均显著高于假伤组(<0.05或<0.01)。12%TBSA全层烧伤组大鼠第一腰椎组织中c-Fos和c-Src的mRNA表达水平显著高于24%TBSA全层烧伤组(<0.01)。重度烧伤可导致大鼠新骨组织生成减少、破骨细胞数量增加和血清磷离子浓度升高,血清P1NP水平降低。骨组织中骨保护素、RANKL、TRAF-6、NFATC1、c-Fos和c-Src的水平呈上升趋势,而NFATC1、c-Fos和c-Src的水平随烧伤面积增加呈下降趋势。

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Vitamin D in burn-injured patients.烧伤患者体内的维生素D
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