脊髓水平Caspase-1活化在大鼠急性切口痛中作用的研究
发布时间：2018-10-14 11:28
【摘要】：第一部分急性切口痛模型大鼠脊髓caspase-1及下游促炎性因子IL-1β表达变化目的:建立大鼠急性切口痛模型,检测急性切口痛模型大鼠脊髓半胱氨酸天冬氨酸蛋白水解酶-1(caspase-1)及下游促炎性因子IL-1β表达变化。方法:雄性SD(Sprague Dawley)大鼠240±10g,36只,随机分为两组(n=18):切口痛组(I组),对照组(C组),每组18只,I组参照文献在大鼠左后足建立切口痛模型,C组不做任何处理。应用von Frey测痛仪测定大鼠的机械缩足反应阈值(paw mechanical withdrawal threshold,PMWT),分别检测大鼠基础PMWT值(T0)、切口痛模型建立后2h(T_1)、6h(T_2)、24h(T_3)、48h(T_4)的PMWT值,最后一次检测完PMWT后,每组随机抽取6只大鼠,取脊髓L4-6。分别采用RT-PCR检测caspase-1mRNA表达、免疫组织化学法检测caspase-1(p20)蛋白表达、酶联免疫吸附法(ELLSA)检测IL-1β蛋白表达。结果:两组大鼠基础PMWT值比较差异无统计学意义(P0.05);I组大鼠T_1、T_2、T_3、T_4、各时点PMWT值比C组明显降低,差异有统计学意义(P0.05);I组T_1、T_2、T_3、T_4、各时点比T0点PMWT值明显降低,差异有统计学意义(P0.05);I组caspase-1mRNA、caspase-1(p20)蛋白、IL-1β蛋白表达均高于C组(P0.05)。第二部分鞘内注射caspase-1抑制剂对急性切口痛模型大鼠的影响目的:通过鞘内注射caspase-1抑制剂Ac-YVAD-CMK,检测其对急性切口痛模型大鼠疼痛阈值及促炎性因子IL-1β表达的影响,探讨caspase-1活化在急性切口痛形成中的作用。方法:鞘内置管成功的雄性SD(Sprague Dawley)大鼠240±10g,18只,随机分为三组(n=6):切口痛组(I组)、切口痛+溶剂组(IR组)、切口痛+拮抗剂组(IJ组),I组鞘内注射生理盐水再建立切口痛模型、IR组建立切口痛模型前鞘内注射溶剂(DMSO)、IJ组建立切口痛模型前鞘内注射caspase-1抑制剂Ac-YVAD-CMK,三组大鼠分别在建立模型前鞘内注射生理盐水、DMSO、Ac-YVAD-CMK 20ul每只,每日一次。分别检测大鼠鞘内置管前、后PMWT值(T0)、(T_1)、切口痛模型建立后2h(T_2)、6h(T_3)、24h(T_4)48h(T_5)的PMWT值,最后一次检测完PMWT后,取大鼠脊髓L4-6,酶联免疫吸附法(ELLSA)检测IL-1β蛋白表达。结果:三组大鼠基础PMWT值、鞘内置管前后PMWT值比较差异无统计学意义(P0.05);I组与IR组在T_2、T_3、T_4、T_5各时点PMWT值比较差异无统计学意义(P0.05);IJ组在T_2、T_3、T_4、T_5各时点PMWT值比I组升高,差异有统计学意义(P0.05);I组与IR组比IL-1β蛋白表达差异无统计意义(P0.05);IJ组与I组比较IL-1β蛋白表达降低(P0.05)结论:急性切口痛模型大鼠脊髓caspase-1被激活,催化其下游促炎性因子IL-1β表达增加。脊髓水平caspase-1活化参与了急性切口痛大鼠中枢敏化的过程。
[Abstract]:The first part of the acute incision pain model rats spinal cord caspase-1 and downstream pro-inflammatory factor IL-1 尾 expression changes objective: to establish acute incision pain model in rats, The changes of spinal cord cysteine aspartate proteolytic enzyme 1 (caspase-1) and downstream pro-inflammatory factor (IL-1 尾) were detected in acute incision pain model rats. Methods: 36 male SD (Sprague Dawley) rats were randomly divided into two groups (n = 18): incision pain group (group I) and control group (group C). The mechanical foot contraction threshold (paw mechanical withdrawal threshold,PMWT) of rats was measured by von Frey. The basic PMWT value (T0), the PMWT values of T 1, T 2, T 3, T 3 and T 4 were measured respectively. After PMWT was last detected, 6 rats in each group were randomly selected for L4-6. The expression of caspase-1mRNA was detected by RT-PCR, the expression of caspase-1 (p20) was detected by immunohistochemical method, and the expression of IL-1 尾 by (ELLSA) was detected by enzyme-linked immunosorbent assay (ELLSA). Results: there was no significant difference in basic PMWT between the two groups (P0.05) in the); I group (P0.05). The PMWT value of the two groups was significantly lower than that of the C group at each time point (P0.05) in the); I group, and the PMWT value at each time point was significantly lower than that in the T0 point. The expression of caspase-1mRNA,caspase-1 (p20) protein and IL-1 尾 protein in); I group were significantly higher than those in C group (P0.05). The second part was the effect of intrathecal injection of caspase-1 inhibitor on acute incision pain model rats. Objective: to investigate the effect of intrathecal injection of caspase-1 inhibitor Ac-YVAD-CMK, on the pain threshold and the expression of pro-inflammatory factor IL-1 尾 in acute incision pain model rats. To investigate the role of caspase-1 activation in the formation of acute incision pain. Methods: 18 male SD (Sprague Dawley) rats with successful intrathecal catheterization were used. They were randomly divided into three groups: incision pain group (group I), incision pain solvent group (IR group), incision pain antagonist group (IJ group,), I group) and intrathecal injection of normal saline to establish incision pain model. IR group established incision pain model by intrathecal injection of solvent (DMSO), IJ before incision pain model. Three groups of rats with intrathecal injection of caspase-1 inhibitor Ac-YVAD-CMK, were injected with normal saline before the establishment of the model of incision pain. Each of them was injected with DMSO,Ac-YVAD-CMK 20ul. Once a day. The PMWT values of anterior and posterior PMWT (T _ 0), (_ T _ 1), 2 h (T _ 2), 6 h (T _ 3), and 24 h (T _ 4) 48 h (T _ 5) PMWT values of rat spinal cord were measured respectively. After PMWT, L4-6 of rat spinal cord was collected and the expression of IL-1 尾 protein was detected by enzyme-linked immunosorbent assay (ELLSA). Results: there was no significant difference in basic PMWT value, PMWT value before and after placement of intrathecal tube between the three groups (P0.05). There was no significant difference in PMWT value between); I group and IR group at T _ 2T _ (3) T _ (3) T _ (4) T _ (5) (P0.05) the PMWT value in); IJ group was higher than that in group I at each time point. There was no significant difference in the expression of IL-1 尾 protein between); I group and IR group (P 0.05). The expression of IL-1 尾 protein in group I was lower than that in group); IJ (P0.05). Conclusion: the expression of caspase-1 in spinal cord of acute incision pain model rats was activated and the expression of IL-1 尾 was increased in the downstream of); IJ group and IR group. The activation of caspase-1 at spinal cord level is involved in the process of central sensitization in acute incision pain rats.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R614
[Abstract]:The first part of the acute incision pain model rats spinal cord caspase-1 and downstream pro-inflammatory factor IL-1 尾 expression changes objective: to establish acute incision pain model in rats, The changes of spinal cord cysteine aspartate proteolytic enzyme 1 (caspase-1) and downstream pro-inflammatory factor (IL-1 尾) were detected in acute incision pain model rats. Methods: 36 male SD (Sprague Dawley) rats were randomly divided into two groups (n = 18): incision pain group (group I) and control group (group C). The mechanical foot contraction threshold (paw mechanical withdrawal threshold,PMWT) of rats was measured by von Frey. The basic PMWT value (T0), the PMWT values of T 1, T 2, T 3, T 3 and T 4 were measured respectively. After PMWT was last detected, 6 rats in each group were randomly selected for L4-6. The expression of caspase-1mRNA was detected by RT-PCR, the expression of caspase-1 (p20) was detected by immunohistochemical method, and the expression of IL-1 尾 by (ELLSA) was detected by enzyme-linked immunosorbent assay (ELLSA). Results: there was no significant difference in basic PMWT between the two groups (P0.05) in the); I group (P0.05). The PMWT value of the two groups was significantly lower than that of the C group at each time point (P0.05) in the); I group, and the PMWT value at each time point was significantly lower than that in the T0 point. The expression of caspase-1mRNA,caspase-1 (p20) protein and IL-1 尾 protein in); I group were significantly higher than those in C group (P0.05). The second part was the effect of intrathecal injection of caspase-1 inhibitor on acute incision pain model rats. Objective: to investigate the effect of intrathecal injection of caspase-1 inhibitor Ac-YVAD-CMK, on the pain threshold and the expression of pro-inflammatory factor IL-1 尾 in acute incision pain model rats. To investigate the role of caspase-1 activation in the formation of acute incision pain. Methods: 18 male SD (Sprague Dawley) rats with successful intrathecal catheterization were used. They were randomly divided into three groups: incision pain group (group I), incision pain solvent group (IR group), incision pain antagonist group (IJ group,), I group) and intrathecal injection of normal saline to establish incision pain model. IR group established incision pain model by intrathecal injection of solvent (DMSO), IJ before incision pain model. Three groups of rats with intrathecal injection of caspase-1 inhibitor Ac-YVAD-CMK, were injected with normal saline before the establishment of the model of incision pain. Each of them was injected with DMSO,Ac-YVAD-CMK 20ul. Once a day. The PMWT values of anterior and posterior PMWT (T _ 0), (_ T _ 1), 2 h (T _ 2), 6 h (T _ 3), and 24 h (T _ 4) 48 h (T _ 5) PMWT values of rat spinal cord were measured respectively. After PMWT, L4-6 of rat spinal cord was collected and the expression of IL-1 尾 protein was detected by enzyme-linked immunosorbent assay (ELLSA). Results: there was no significant difference in basic PMWT value, PMWT value before and after placement of intrathecal tube between the three groups (P0.05). There was no significant difference in PMWT value between); I group and IR group at T _ 2T _ (3) T _ (3) T _ (4) T _ (5) (P0.05) the PMWT value in); IJ group was higher than that in group I at each time point. There was no significant difference in the expression of IL-1 尾 protein between); I group and IR group (P 0.05). The expression of IL-1 尾 protein in group I was lower than that in group); IJ (P0.05). Conclusion: the expression of caspase-1 in spinal cord of acute incision pain model rats was activated and the expression of IL-1 尾 was increased in the downstream of); IJ group and IR group. The activation of caspase-1 at spinal cord level is involved in the process of central sensitization in acute incision pain rats.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R614
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