TGF-β1、Smad2及Smad3在脊柱结核诊疗中的价值

The value of TGF-β1, Smad2 and Smad3 in the diagnosis and efficacy evaluation of spinal tuberculosis

中文关键词:  脊柱结核  转化生长因子β1  Smad2  Smad3  诊断及疗效评价

英文关键词:Spinal tuberculosis  TGF-β1  Smad2  Smad3  Diagnosis and efficacy evaluation

中文摘要:

  【摘要】 目的：研究转化生长因子β1（transforming growth factor-β1，TGF-β1）、Smad2（drosophila mothers against decapentaplegic 2）及Smad3在脊柱结核诊断及疗效评价中的价值。方法：纳入2018年9月~2019年6月在宁夏医科大学总医院治疗的确诊为脊柱结核（观察组）和椎间盘退变（对照组）患者各35例，分析两组患者临床资料并收集椎间盘病变组织，同时收集观察组术前、术后6个月和术后1年（分别定义为A组、B组和C组）及对照组术前的外周血标本。提取椎间盘组织及外周血标本的RNA，荧光定量PCR法扩增TGF-β1、Smad2、Smad3的mRNA，免疫组化染色法检测其在椎间盘组织中的表达水平，ELISA法测定其在外周血中的蛋白表达水平。t检验分析各指标在组间及组内的表达差异；ROC曲线分析各指标单独及分别联合ESR、CRP诊断脊柱结核的价值；曲线回归分析各指标与脊柱结核患者临床资料的相关性。结果：在椎间盘mRNA水平，观察组TGF-β1、Smad2及Smad3表达高于对照组（P＜0.05）。在椎间盘蛋白水平，观察组TGF-β1、Smad2及Smad3均为阳性表达，对照组均为阴性表达。在外周血mRNA水平，A组TGF-β1和Smad3表达高于对照组，B组表达低于A组（P＜0.01），C组表达低于B组（P＞0.05）；Smad2表达水平在组间及组内比较时均为P＞0.05。在外周血蛋白水平，A组TGF-β1表达高于对照组（P＜0.01），B组表达低于A组（P＜0.01），C组表达低于B组（P＞0.05）；Smad2和Smad3表达水平在组间及组内比较时均为P＞0.05。ROC曲线显示外周血TGF-β1 mRNA、TGF-β1蛋白、Smad3 mRNA单独及分别联合ESR、CRP诊断脊柱结核的AUC分别是0.7800、0.8000、0.8352、0.8219、0.8705、0.8819、0.7410、0.7552、0.8000，敏感性分别是93.33%、73.33%、86.67%、70.00%、90.00%、80.00%、46.67%、46.67%、80.00%，特异性分别是54.29%、80.00%、80.00%、91.43%、88.57%、94.29%、100.00%、100.00%、85.71%（均为P＜0.01）。曲线回归分析表明外周血TGF-β1和Smad3在mRNA水平表达呈显著正相关（R2=0.8534，P＜0.01），外周血TGF-β1蛋白表达水平与脊柱结核患者的年龄、病程、病变椎体数量、ESR、CRP并无相关性。结论：TGF-β1/Smads信号通路与脊柱结核发病相关，其中TGF-β1/Smad3可能发挥主要作用；外周血及椎间盘组织TGF-β1、Smad3 mRNA检测可用于辅助脊柱结核诊断及疗效评价；外周血TGF-β1蛋白高表达，且与预后相关，可作为脊柱结核的诊断及疗效评价指标，与CRP联合诊断时价值更高。

英文摘要:

  【Abstract】 Objectives: To study the value of transforming growth factor-β1(TGF-β1), Smad2 and Smad3 in the diagnosis and efficacy evaluation of spinal tuberculosis. Methods: Thirty-five patients diagnosed with spinal tuberculosis and treated in General Hospital of Ningxia Medical University from September 2018 to June 2019 were enrolled in the experimental group, and another thirty-five patients diagnosed with degenerative intervertebral disc and treated in the same hospital during the same period were enrolled in the control group. The clinical information of patients in both groups were analyzed, and diseased tissues of intervertebral disc of each group of patients were collected. Besides, 35 specimens of peripheral blood of patients in the experimental group were collected before operation(group A), 6 months after operation(group B) and 1 year after operation(group C). And 35 specimens of preoperative peripheral blood in the control group were also collected. The RNA from tissues of intervertebral disc and specimens of peripheral blood were extracted, then the concentration of mRNA of TGF-β1, Smad2 and Smad3 was amplified via PCR, and the mRNA expression levels in intervertebral disc were detected via immunohistochemical staining while the expression levels of protein in blood were measured via ELISA. At last, the intragroup and intergroup differences in expressions were analyzed through t test, the value of independent indexes and their combined value with ESR and CRP in the diagnosis of spinal tuberculosis were analyzed by ROC curve, and the correlations of the three indexes with clinical data of patients were evaluated by curve regression. Results: The mRNA of TGF-β1, Smad2 and Smad3 in experimental group was higher than that of control group from the disc(P<0.05). TGF-β1, Smad2 and Smad3 were positively expressed in the experimental group, and negatively expressed in the control group at the level of intervertebral disc protein. In the peripheral blood, mRNA of TGF-β1 and Smad3 in group A was higher than that in the control group, and it in group B was lower than that in group A(P<0.01), in group C it was lower than that in group B(P＜0.05), but Smad2 had no difference within groups(P>0.05). At the protein level, TGF-β1 in group A was higher than that in the control group(P<0.01), and it in group B was lower than that in group A(P<0.01), and which was lower in group C than that in group B(P＜0.05), but Smad2 and Smad3 had no difference within groups(P>0.05). The AUC of TGF-β1 mRNA, TGF-β1 protein and Smad3 mRNA in blood alone and combined with ESR or CRP respectively were 0.7800, 0.8000, 0.8352, 0.8219, 0.8705, 0.8819, 0.7410, 0.7552 and 0.8000, the sensitivity were 93.33%, 73.33%, 86.67%, 70.00%, 90.00%, 80.00%, 46.67%, 46.67% and 80.00%, respectively, the specificity were 54.29%, 80.00%, 80.00%, 91.43%, 88.57%, 94.29%, 100.00%, 100.00% and 85.71%(P<0.01), respectively. The mRNA of TGF-β1 and Smad3 had positive correlation(R2=0.8534, P<0.01), but it was not correlated with age, course of disease, number of diseased vertebrae, ESR and CRP. Conclusions: The TGF-β1/Smads pathway participates the onset of spinal tuberculosis, and TGF-β1/Smad3 may play a major role. The detection of TGF-β1 and Smad3 mRNA in peripheral blood and intervertebral disc tissue can be used to assist the diagnosis and efficacy evaluation of spinal tuberculosis. The expression of TGF-β1 protein in peripheral blood is highly expressed and related to prognosis. It can be used as an index for the diagnosis and efficacy evaluation of spinal tuberculosis, and it′s more valuable when combined with CRP.