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目的:观察高迁移率族蛋白-1(high mobility group box-1,HMGB1)在糖尿病大鼠脊髓内的表达变化,探索其参与糖尿病性机械性痛觉过敏的具体机制,进一步阐明糖尿病性痛的机制,为糖尿病疼痛的治疗提供新的思路。方法:(1)36只SD大鼠随机分成6组(n=6),分别为正常大鼠组、糖尿病大鼠对照组、糖尿病7 d组、14 d、21 d和28 d组。通过Real-time PCR法检测各组大鼠脊髓内HMGB1 m RNA的表达情况。(2)24只SD大鼠分成4组(n=6)制作糖尿病大鼠模型,在造模后第28 d鞘内给予生理盐水、HMGB1的中和抗体10、30和100μg,检测糖尿病大鼠模型在各时间点的机械性缩足阈值。(3)30只SD大鼠随机分成5组(n=6),其中4组给予链尿佐菌素制作糖尿病大鼠模型。模型制作28 d后鞘内给予生理盐水、HMGB1的中和抗体10、30和100μg。另一组大鼠腹腔给予生理盐水,作为糖尿病大鼠的对照组。检测各组大鼠脊髓的TNF-α、IL-1β和IL-6 m RNA的表达。结果:(1)糖尿病大鼠模型制作21 d和28 d,脊髓内HMGB1 m RNA的表达显著上调(P<0.05)。(2)糖尿病大鼠鞘内给予HMGB1中和抗体30和100μg后,可以在长达24 h的时间内扭转模型大鼠的机械性痛敏(P<0.05)。(3)糖尿病大鼠造模28 d后,鞘内给予HMGB1的中和抗体30和100μg可以明显逆转糖尿病大鼠脊髓内的TNF-α、IL-1β和IL-6 m RNA的表达(P<0.05)。结论:糖尿病大鼠脊髓内HMGB1显著上调,鞘内给予HMGB1的中和抗体可以通过抑制脊髓内TNF-α等细胞因子的表达而扭转糖尿病大鼠的机械性痛敏。以上结果提示,脊髓HMGB1可能参与了糖尿病机械性痛敏状态的维持过程。我们的研究对脊髓HMGB1参与糖尿病大鼠的疼痛的机制进行初步的探讨,为糖尿病性痛的治疗提供新的思路。
OBJECTIVE: To investigate the expression of high mobility group box-1 (HMGB1) in the spinal cord of diabetic rats and to explore its mechanism of involvement in diabetic mechanical hyperalgesia and further elucidate the mechanism of diabetic pain , Provide new ideas for the treatment of diabetic pain. Methods: (1) Thirty-six SD rats were randomly divided into 6 groups (n = 6), normal control group, diabetic control group, diabetic 7 d group, 14 d, 21 d and 28 d groups. Real-time PCR was used to detect the expression of HMGB1 mRNA in the spinal cord of each group. (2) Twenty-four SD rats were divided into 4 groups (n = 6) to make a diabetic rat model. At the 28th day after modeling, NS, 10, 30 and 100μg neutralizing antibodies to HMGB1 were given intrathecally. Mechanical contraction threshold of the model at each time point. (3) Thirty SD rats were randomly divided into five groups (n = 6). Four groups were given streptozotocin to make diabetic rat model. After 28 days of modeling, normal saline, 10, 30 and 100 μg of neutralizing antibodies to HMGB1, were administered intrathecally. Another group of rats were given intraperitoneal saline, as a control group of diabetic rats. The expression of TNF-α, IL-1β and IL-6 m RNA in the spinal cord of each group was detected. Results: (1) The expression of HMGB1 m RNA in the spinal cord was significantly up-regulated (P <0.05) on day 21 and day 28 in diabetic rats. (2) After 30 and 100 μg of HMGB1 neutralizing antibody were intrathecally administered to diabetic rats, the mechanical hyperalgesia of model rats could be reversed within 24 hours (P <0.05). (3) After 28 days of modeling, the neutralizing antibody of HMGB1 administered intrathecally could obviously reverse the expression of TNF-α, IL-1β and IL-6 m RNA in the spinal cord of diabetic rats (P < 0.05). CONCLUSIONS: HMGB1 in the spinal cord of diabetic rats was significantly up-regulated. Neutralizing antibody of HMGB1 administered intrathecally could reverse the mechanical hyperalgesia of diabetic rats by inhibiting the expression of TNF-α and other cytokines in the spinal cord. These results suggest that spinal cord HMGB1 may be involved in the maintenance of diabetic mechanical pain-sensitive state. Our study preliminarily explored the mechanism of the involvement of spinal cord HMGB1 in the pain of diabetic rats, providing new ideas for the treatment of diabetic pain.