脊髓水平Caspase-1活化在大鼠急性切口痛中作用的研究
發(fā)布時(shí)間：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表達(dá)變化目的:建立大鼠急性切口痛模型,檢測(cè)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表達(dá)變化。方法:雄性SD(Sprague Dawley)大鼠240±10g,36只,隨機(jī)分為兩組(n=18):切口痛組(I組),對(duì)照組(C組),每組18只,I組參照文獻(xiàn)在大鼠左后足建立切口痛模型,C組不做任何處理。應(yīng)用von Frey測(cè)痛儀測(cè)定大鼠的機(jī)械縮足反應(yīng)閾值(paw mechanical withdrawal threshold,PMWT),分別檢測(cè)大鼠基礎(chǔ)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,最后一次檢測(cè)完P(guān)MWT后,每組隨機(jī)抽取6只大鼠,取脊髓L4-6。分別采用RT-PCR檢測(cè)caspase-1mRNA表達(dá)、免疫組織化學(xué)法檢測(cè)caspase-1(p20)蛋白表達(dá)、酶聯(lián)免疫吸附法(ELLSA)檢測(cè)IL-1β蛋白表達(dá)。結(jié)果:兩組大鼠基礎(chǔ)PMWT值比較差異無統(tǒng)計(jì)學(xué)意義(P0.05);I組大鼠T_1、T_2、T_3、T_4、各時(shí)點(diǎn)PMWT值比C組明顯降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05);I組T_1、T_2、T_3、T_4、各時(shí)點(diǎn)比T0點(diǎn)PMWT值明顯降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05);I組caspase-1mRNA、caspase-1(p20)蛋白、IL-1β蛋白表達(dá)均高于C組(P0.05)。第二部分鞘內(nèi)注射caspase-1抑制劑對(duì)急性切口痛模型大鼠的影響目的:通過鞘內(nèi)注射caspase-1抑制劑Ac-YVAD-CMK,檢測(cè)其對(duì)急性切口痛模型大鼠疼痛閾值及促炎性因子IL-1β表達(dá)的影響,探討caspase-1活化在急性切口痛形成中的作用。方法:鞘內(nèi)置管成功的雄性SD(Sprague Dawley)大鼠240±10g,18只,隨機(jī)分為三組(n=6):切口痛組(I組)、切口痛+溶劑組(IR組)、切口痛+拮抗劑組(IJ組),I組鞘內(nèi)注射生理鹽水再建立切口痛模型、IR組建立切口痛模型前鞘內(nèi)注射溶劑(DMSO)、IJ組建立切口痛模型前鞘內(nèi)注射caspase-1抑制劑Ac-YVAD-CMK,三組大鼠分別在建立模型前鞘內(nèi)注射生理鹽水、DMSO、Ac-YVAD-CMK 20ul每只,每日一次。分別檢測(cè)大鼠鞘內(nèi)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,最后一次檢測(cè)完P(guān)MWT后,取大鼠脊髓L4-6,酶聯(lián)免疫吸附法(ELLSA)檢測(cè)IL-1β蛋白表達(dá)。結(jié)果:三組大鼠基礎(chǔ)PMWT值、鞘內(nèi)置管前后PMWT值比較差異無統(tǒng)計(jì)學(xué)意義(P0.05);I組與IR組在T_2、T_3、T_4、T_5各時(shí)點(diǎn)PMWT值比較差異無統(tǒng)計(jì)學(xué)意義(P0.05);IJ組在T_2、T_3、T_4、T_5各時(shí)點(diǎn)PMWT值比I組升高,差異有統(tǒng)計(jì)學(xué)意義(P0.05);I組與IR組比IL-1β蛋白表達(dá)差異無統(tǒng)計(jì)意義(P0.05);IJ組與I組比較IL-1β蛋白表達(dá)降低(P0.05)結(jié)論: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表達(dá)增加。脊髓水平caspase-1活化參與了急性切口痛大鼠中樞敏化的過程。
[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.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：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.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R614
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