加味蠲痹汤对腰椎后路减压融合内固定术后椎旁肌损伤模型SD大鼠影响实验研究

Effect of Modified Zhibi Decoction for SD Rats Model of Paravertebral Muscle Injury after Lumbar Posterior Decompression-fusion and Internal Fixation

目的:探究加味蠲痹汤对腰椎后路减压融合内固定术后椎旁肌损伤模型SD大鼠疗效及碱性成纤维细胞生长因子(Basic Fibroblast Growth Factor,bFGF)mRNA、Ⅰ型胶原蛋白(Collagen Ⅰ)、Ⅲ型胶原蛋白(Collagen Ⅲ)的作用。方法:大鼠均模拟临床腰椎后路减压融合内固定术模式建立椎旁肌损伤模型。将15只建模成功的SD大鼠随机分为3组(n=5),即模型组、低剂量组和高剂量组,另取5只健康大鼠作为对照组。低剂量组和高剂量组的大鼠分别按照9.1g/Kg、18.2g/Kg体重标准使用加味蠲痹汤灌胃。干预5d后椎旁肌损伤评分和苏木素-伊红(HE)染色分析损伤恢复情况。检测和比较血清中组胺和肿瘤坏死因子α(Tumor Necrosis Factor-α,TNF-α)水平。使用qPCR和Western blot检测软组织中bFGF mRNA、Collagen-Ⅰ/Ⅲ蛋白的水平。结果:低剂量组和高剂量组大鼠椎旁肌损伤评分低于模型组(P<0.05),并且高剂量组的椎旁肌损伤评分低于低剂量组(P<0.05)。HE染色结果显示对照组的软组织结构正常;模型组细胞排列紊乱、纤维断裂和炎性浸润,同时出现纤维增生;低剂量组和高剂量组的椎旁肌损伤程度均显著轻于模型组,并且高剂量组的软组织基本恢复。模型组大鼠血清组胺和TNF-α的水平明显升高(P<0.05),与模型组比较,低剂量组和高剂量组的水平显著降低(P<0.05),其中高剂量组大鼠血清组胺和TNF-α的水平显著低于低剂量组(P<0.05)。模型组大鼠软组织中bFGF mRNA、Collagen-Ⅰ/Ⅲ蛋白水平明显升高(P<0.05),与模型组比较,低剂量组和高剂量组bFGF mRNA显著升高而Collagen-Ⅰ/Ⅲ蛋白的水平显著降低(P<0.05),其中高剂量组大鼠软组织中bFGF mRNA显著高于低剂量组,而Collagen-Ⅰ/Ⅲ蛋白水平显著低于低剂量组(P<0.05)。结论:加味蠲痹汤可有效的促进腰椎后路减压融合内固定术后椎旁肌损伤大鼠的组织恢复,降低大鼠炎症反应,增加bFGF mRNA蛋白表达,降低Collagen-Ⅰ/Ⅲ蛋白表达。

Objective:Investigate the therapeutic effect of Modified Zhibi Decoction on SD rats model of paravertebral injury after posterior decompression and fusion of lumbar spine and the role of basic fibroblast growth factor(bFGF)mRNA,collagen type Ⅰ(Collagen Ⅰ),collagen Ⅲ(Collagen Ⅲ).Methods:The rats were modeled by the clinical model of posterior lumbar spine decompression and fusion and internal fixation.Fifteen SD rats successfully modeled were randomly divided into three groups(n=5),namely the model group,the low-dose group and the high-dose group,and five healthy rats were selected as the controlled group.The rats in the low-dose group and the high-dose group were administered with Modified Zhibi Decoction according to the body weight standards of 9.1g/kg and 18.2g/kg,respectively.After 5days of intervention,the paraspinal muscle injury score and hematoxylineosin(HE)staining were used to analyze the injury recovery.Detection and comparison of serum histamine and tumor necrosis factor-α(TNF-α)levels.The levels of bFGF mRNA and Collagen-Ⅰ/Ⅲ protein in soft tissue were detected by qPCR and Western blot.Results:The low-dose and high-dose groups had lower paraspinal muscle injury scores than the model group(P<0.05),and the high-dose group had lower paraspinal muscle injury scores than the low-dose group(P<0.05).The results of HE staining showed that the soft tissue structure of the controlled group was normal.The cell arrangement disorder,fiber breakage,and inflammatory infiltration in the model group were accompanied by fibroproliferation.The degree of paraspinal muscle injury was significantly less in the low and high dose groups than in the model group,and the soft tissue of the high-dose group recovered basically.The levels of serum histamine and TNF-α in the model group were significantly increased(P<0.05).Compared with the model group,the levels of the low-dose group and the high-dose group were significantly reduced(P<0.05).The levels of serum histamine and TNF-α were significantly lower than those in the low-dose group(P<0.05).The levels of bF-GF mRNA and Collagen-Ⅰ/Ⅲ protein in the soft tissues of the model group rats were significantly increased(P<0.05).Compared with the model group,the bFGF mRNA was significantly increased in the low-dose group and the high-dose group,while the level of collagen was significantly reduced(P<0.05).The bFGFmRNA in the soft tissue of rats in the high-dose group was significantly higher than that in the low-dose group and the level of Collagen-Ⅰ/Ⅲprotein was significantly lower than that in the low-dose group(P<0.05).Conclusion:Modified Zhibi Decoction can effectively promote the tissue recovery of paravertebral muscle injury rats after posterior lumbar decompression fusion and internal fixation,reduce the inflammatory response in rats,increase the expression of bFGF mRNA protein,and reduce the expression of Collagen-Ⅰ/Ⅲ protein.