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目的研究坐骨神经选择性损伤引起的慢性疼痛应激态下SD大鼠颌下腺中annexin 5表达的变化。方法72只SD大鼠随机均分为3组:手术组、假手术组和对照组(每组24只)。手术组:用氯胺酮(40mg/kg)腹腔注射麻醉后,切开左后肢侧面皮肤和股二头肌,暴露大鼠坐骨神经干及其3个分支:胫神经、腓总神经和腓肠神经,结扎并切断胫神经和腓总神经,保留腓肠神经以建立坐骨神经分支选择性损伤(SNI)模型。假手术组:只暴露坐骨神经干及其分支而不损伤神经。对照组:不做任何处理。分别于手术后14、21、28d从各组中取8只大鼠,麻醉后处死,分离颌下腺组织置于液氮内保存备用。然后通过免疫组织化学和Western blotting检测坐骨神经选择性损伤后14、21、28d时各组大鼠颌下腺中annexin 5蛋白定位及表达的变化。结果免疫组化结果显示,annexin 5主要定位在颌下腺润管、分泌管及排泄导管上皮细胞中。术后14d,annexin 5在颌下腺的定位明显减少,Western blotting结果显示术后14d时与对照组比较,手术组大鼠颌下腺中annexin 5表达水平显著降低(P<0.01),与假手术组比较,手术组annexin 5表达水平也显著下降(P<0.05)。但术后21和48d手术组与对照组、假手术组比较,差异无统计学意义(P>0.05)。结论坐骨神经选择性损伤引起的慢性疼痛应激能下调SD大鼠颌下腺中annexin 5的表达,annexin 5可能参与了颌下腺对应激的反应。
Objective To study the changes of annexin 5 expression in the submandibular gland of SD rats under chronic pain stress induced by selective sciatic nerve injury. Methods 72 SD rats were randomly divided into three groups: operation group, sham operation group and control group (24 rats in each group). Surgical group: After anesthesia was induced by ketamine (40 mg / kg), lateral skin and biceps femoris of the left hind limbs were excised and the sciatic nerve trunk of the rat and its three branches were exposed: the tibial nerve, the common peroneal nerve and the sural nerve, The tibial nerve and common peroneal nerve were severed, and the sural nerve was preserved to establish the sciatic nerve branch selective injury (SNI) model. Sham-operated group: only exposing the sciatic nerve trunk and its branches without damaging the nerve. Control group: without any treatment. At the 14th, 21st and 28th day after surgery, 8 rats from each group were taken and anesthetized. After sacrificed, the submandibular glands were separated and stored in liquid nitrogen for later use. Immunohistochemistry and Western blotting were used to detect the localization of annexin 5 protein in rat submandibular glands at 14, 21 and 28 days after sciatic nerve selective injury. Results Immunohistochemical results showed that annexin 5 mainly located in submandibular gland duct, secretory duct and excretory duct epithelial cells. The expression of annexin 5 in the submandibular gland of the operation group was significantly lower than that of the control group on day 14 postoperatively (P <0.01). The level of annexin 5 in the submandibular gland of the operation group was significantly lower than that of the sham operation group The expression of annexin 5 in the operation group also decreased significantly (P <0.05). However, there was no significant difference between the operation group and the control group and the sham operation group 21 and 48 days after operation (P> 0.05). Conclusion Chronic pain stress induced by selective sciatic nerve injury can down-regulate the expression of annexin 5 in submandibular gland of SD rats. Annexin 5 may be involved in the response of submandibular gland to stress.