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西南医科大学附属中医医院疼痛科,泸州 646000
廖 瑶,Email:liaoyao202110@163.com,研究方向:疼痛机制与治疗研究。
收稿:2022-06-22,
修回:2023-04-25,
录用:2023-05-23,
纸质出版:2023-07-28
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廖瑶,涂婷,曾惜羽,等.瑞芬太尼对椎间盘退变模型大鼠椎间盘的影响及机制[J].重庆医科大学学报,2023,48(7):760-764.
Liao Yao,Tu Ting,Zeng Xiyu,et al.Effect of remifentanil on the intervertebral disc of rats with intervertebral disc degeneration and its mechanism[J].Journal of Chongqing Medical University,2023,48(7):760-764.
廖瑶,涂婷,曾惜羽,等.瑞芬太尼对椎间盘退变模型大鼠椎间盘的影响及机制[J].重庆医科大学学报,2023,48(7):760-764. DOI: 10.13406/j.cnki.cyxb.003281.
Liao Yao,Tu Ting,Zeng Xiyu,et al.Effect of remifentanil on the intervertebral disc of rats with intervertebral disc degeneration and its mechanism[J].Journal of Chongqing Medical University,2023,48(7):760-764. DOI: 10.13406/j.cnki.cyxb.003281.
目的
2
观察瑞芬太尼对椎间盘退变(intervertebral disc degeneration,IDD)模型大鼠椎间盘的保护作用,并探讨其作用机制。
方法
2
将40只健康SD大鼠随机分为假手术组、模型组和瑞芬太尼低、中、高剂量组,每组8只;采用针刺纤维环法构建IDD模型;瑞芬太尼低、中、高剂量组于造模后以0.2、0.6、1.2 μg/(kg·min)经大鼠尾静脉泵注瑞芬太尼30 min,模型组和假手术组给予等量生理盐水泵注。采用苏木精-伊红(hematoxylin-eosin,HE)染色观察椎间盘组织病理变化;酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测血清肿瘤坏死因子-α(tumor necrosis factor α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)水平;Western blot检测椎间盘组织过氧化物酶体增殖物激活受体γ共激活因子-1α(peroxisome proliferator-activated receptor-1α,PGC-1α)、线粒体转录因子(mitochondrial transcription factor A,TFAM)、半胱氨酸天冬氨酸蛋白酶1(cysteine aspartic protease-1,Caspase-1)、消皮素D(gasdermin D,GSDMD)、NOD样受体热蛋白结构域3(NOD-like receptor family pyrin domain containing 3,NLRP3)、缺氧诱导因子-1α(hypoxia-inducible factor 1α,HIF-1α)、血管内皮生长因子(vascular endothelial growth factor,VEGF)蛋白表达。
结果
2
与假手术组比较,模型组大鼠髓核缩小,纤维环排列紊乱,髓核细胞数量减少;血清TNF-α、IL-6、IL-1β水平升高(
P
<
0.05);椎间盘组织PGC-1ɑ、TFAM蛋白表达水平降低,Ca
spase-1、GSDMD、NLRP3、HIF-1α、VEGF蛋白表达水平增加(
P
<
0.05)。与模型组比较,瑞芬太尼中、高剂量组纤维环排列紊乱明显改善,髓核细胞数量明显增多;血清TNF-α、IL-6、IL-1β水平降低(
P
<
0.05);椎间盘组织PGC-1α、TFAM蛋白表达水平增加,Caspase-1、GSDMD、NLRP3、HIF-1α、VEGF蛋白表达水平降低(
P
<
0.05)。
结论
2
瑞芬太尼可改善IDD模型大鼠椎间盘退变,其机制与抑制椎间盘炎症反应、细胞焦亡,调控HIF-1α/VEGF信号通路有关。
Objective
2
To observe the protective effect of remifentanil on intervertebral disc of rats with intervertebral disc degeneration (IDD),and to explore its mechanism of action.
Methods
2
A total of 40 healthy SD rats were randomly divided into sham-operation group,model group,low-dose,middle-dose and high-dose remifentanil groups,8 cases in each group. IDD models were constructed by fibrous annulus puncturing. After modeling,low-dose,medium-dose and high-dose remifentanil groups were given intravenous injection of remifentanil through tail vein at rates of 0.2,0.6 and 1.2 μg/(kg·min) for 30 minutes,while model group and sham operation group were given the same volume of normal saline. The pathological changes of intervertebral disc tissues were observed by hematoxylin-eosin(HE) staining. The levels of serum tumor necrosis factor α(TNF-α),interleukin 6(IL-10) and interleukin 1β(IL-1β) were detected by enzyme linked immunosorbent assay(ELISA). The expressions of peroxisome proliferator-activated receptor-1α(PGC-1α),mitochondrial transcription factor A(TFAM),cysteine aspartic protease-1(Caspase-1),gasdermin D(GSDMD),NOD-like receptor family pyrin domain containing 3(NLRP3),hypoxia-inducible factor 1α(HIF-1α) and vascular endothelial growth factor(VEGF) in intervertebral disc tissues were detected by Western blot.
Results
2
Compared with sham operation group,nucleus pulposus was shrunk,arrangement of fibrous annulus was disordered and the number of nucleus pulposus cells was decreased in model group. The levels of serum TNF
-α,IL-6 and IL-1β were increased(
P
<
0.05). The expression levels of PGC-1α and TFAM in intervertebral disc tissues were decreased,and expression levels of Caspase-1,GSDMD,NLRP3,HIF-1α and VEGF were increased(
P
<
0.05). Compared with model group,disorders of fibrous annulus were significantly improved in medium-dose and high-dose remifentanil groups,and number of nucleus pulposus cells was significantly increased. The levels of serum TNF-α,IL-6 and IL-1β were decreased(
P
<
0.05). The expression levels of PGC-1α and TFAM in intervertebral disc tissues were increased,and expression levels of Caspase-1,GSDMD,NLRP3,HIF-1α and VEGF were decreased(
P
<
0.05).
Conclusion
2
Remifentanil can improve IDD in rats,and its mechanism is related to inhibiting inflammatory response and pyroptosis of intervertebral disc,and regulating HIF-1α/VEGF signaling pathways.
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