重组hIL-10抗家兔皮肤移植反应中IL-17家族的变化
发布时间：2018-08-27 15:52
【摘要】：目的：通过观察家兔皮肤移植经rhIL-10作用后,血清中IL-17家族因子的变化,探讨IL-17家族因子在rhIL-10抗免疫排斥机制中的作用,为rhIL-10的开发及应用提供实验依据。 方法：(1)皮肤移植家兔分6组：①rhIL-10低剂量组；②rhIL-10高剂量组；③CsA低剂量组；④CsA高剂量组；⑤rhIL-10+CsA低剂量联合组；⑥生理盐水组。 (2)用药方案：于术前1天开始用药,每只家兔连续10天肌肉注射。 (3)各组分别于术前1天、术后1、4、7、14、21、28天取外周血,分离血清,以酶联免疫吸附法检测移植家兔血清IL-17家族细胞因子水平。 结果：(1)IL-17A检测结果表明,各实验组家兔外周血清IL-17A水平较术前明显升高,在术后14天达到最高值,与术前相比差异有统计学意义(p0.05),然后逐渐降低；生理盐水对照组IL-17A水平比其它组均高,差异有统计学意义(p0.05)；rhIL-10高剂量组IL-17A水平较rhIL-10低剂量组低,差异有统计学意义(p0.05)；联合组比CsA低剂量组、rhIL-10低剂量组均低,且有统计学差异(p0.05)；在术后1天,rhIL-10低剂量组IL-17A水平比CsA低剂量组水平高,且有统计学差异(p0.05),但在高剂量组二者无统计学差异(p0.05),在其它时间点,rhIL-10低剂量组与CsA低剂量组之间、rhIL-10高剂量组与CsA高剂量组之间IL-17A水平无统计学差异(p0.05)。 (2)IL-27的检测结果表明,各实验组家兔外周血清IL-27水平较术前明显升高,在术后14天达到最高值,与术前相比差异有统计学意义(p0.05),然后逐渐降低；生理盐水对照组IL-27水平比其它组均高,且有统计学差异(p0.05)；rhIL-10高剂量组IL-27水平较rhIL-10低剂量组水平低,且有统计学差异(p0.05)；联合组比CsA低剂量组、rhIL-10低剂量组均低,且有统计学差异(p0.05)；在术后1、4、7天rhIL-10低剂量组IL-27水平比CsA低剂量组水平高,且有统计学差异(p0.05),在其它时间,rhIL-10低剂量组IL-27水平与CsA低剂量组相比无统计学差异(p0.05)；rhIL-10高剂量组与CsA高剂量组相比,IL-27平在各检测时间点均无统计学差异(p0.05)。 (3)IL-25的检测结果表明,各实验组家兔外周血清IL-25水平较术前明显升高,生理盐水对照组在术后第4天达到最高值,其它各实验组在术后14天达到最高值,与术前相比差异有统计学意义(p0.05),然后逐渐降低；在术后1天,生理盐水对照组IL-25水平比其它组高但无统计学差异(p0.05),在术后4天,生理盐水对照组IL-25水平比其它组低但无统计学差异(p0.05),从术后7天开始,生理盐水对照组IL-25水平比其它组均低,且有统计学差异(p0.05)；除生理盐水对照组外,其他各组之间IL-25水平均无统计学差异(p0.05)。 (4)IL-17F检测结果表明,各实验组外周血IL-17F水平较术前明显升高,在术后14天达到最高值,与术前相比差异有统计学意义(p0.05),然后逐渐降低；生理盐水对照组IL-17F水平比其它组均高,且有统计学差异(p0.05)；rhIL-10高剂量组IL-17F水平较rhIL-10低剂量组水平低,且有统计学差异(p0.05)；联合组比CsA低剂量组、rhIL-10低剂量组均低,且有统计学差异(p0.05)；在术后1天rhIL-10低剂量组IL-17F水平比CsA低剂量组水平高,且有统计学差异(p0.05),rhIL-10高剂量组IL-17F水平比CsA高剂量组水高,且有统计学差异(p0.05)；在其它时间,rhIL-10低剂量组与CsA低剂量组,rhIL-10高剂量组与CsA高剂量组之间,IL-17水平无统计学差异(p0.05)。 结论：(1)IL-17家族细胞因子在皮肤移植术后明显升高,表明IL-17家族细胞因子可能通过大量分泌,介导移植后的炎症反应,以促进炎性因子分泌从而参与移植排斥反应；(2)血清中IL-17家族细胞因子含量变化可以作为移植排斥的诊断与监测指标：(3)rhIL-10对IL-17A、IL-27、IL-17F的抑制与CsA有协同作用。
[Abstract]:Objective: To observe the changes of IL-17 family factors in serum of rabbits after skin transplantation with rhIL-10, and explore the role of IL-17 family factors in the anti-rejection mechanism of rhIL-10, so as to provide experimental basis for the development and application of rhIL-10.
Methods: (1) Skin transplantation rabbits were divided into 6 groups: (1) rhIL-10 low-dose group; (2) rhIL-10 high-dose group; (3) CsA low-dose group; (4) CsA high-dose group; (6) rhIL-10 + CsA low-dose group; (3) saline group.
(2) medication regimen: 1 days before the operation, each rabbit was injected intramuscularly for 10 consecutive days.
(3) Peripheral blood samples were taken from each group one day before operation and 1,4,7,14,21,28 days after operation. Serum levels of IL-17 family cytokines were detected by enzyme-linked immunosorbent assay.
Results: (1) The results of IL-17A test showed that the levels of IL-17A in peripheral serum of rabbits in each experimental group were significantly higher than those before operation, and reached the highest level at 14 days after operation, which was statistically significant (p0.05), and then gradually decreased. The levels of IL-17A in the saline control group were higher than those in other groups, and the difference was statistically significant (p0.05). The level of IL-17A in the dosage group was lower than that in the low dose group of rhIL-10, and the difference was statistically significant (p0.05); the level of IL-17A in the combined group was lower than that in the low dose group of CsA and the low dose group of rhIL-10 was lower and statistically significant (p0.05); the level of IL-17A in the low dose group of rhIL-10 was higher than that in the low dose group of CsA on the first day after operation, and the difference was statistically significant (p0.05). There was no significant difference in IL-17A levels between rhIL-10 low dose group and CSA low dose group at other time points (p0.05).
(2) The results of IL-27 detection showed that the levels of IL-27 in peripheral serum of rabbits in each experimental group were significantly higher than those before operation, and reached the highest level at 14 days after operation, which was significantly different from those before operation (p0.05), and then gradually decreased; the levels of IL-27 in the control group were higher than those in other groups, and there was a statistically significant difference (p0.05). The level of IL-27 in the combined group was lower than that in the low-dose group of CsA and the low-dose group of rhIL-10 (p0.05). The level of IL-27 in the low-dose group of rhIL-10 was higher than that in the low-dose group of CsA at 1,4,7 days after operation, and the difference was statistically significant (p0.05). There was no significant difference between the levels of IL-27 and CSA low-dose group (p0.05), while there was no significant difference between rhIL-10 high-dose group and CSA high-dose group (p0.05).
(3) The results of IL-25 detection showed that the levels of IL-25 in peripheral serum of rabbits in each experimental group were significantly higher than those before operation, and the levels of IL-25 in normal saline control group reached the highest level on the 4th day after operation, while those in other experimental groups reached the highest level on the 14th day after operation, which was significantly different from those before operation (p0.05), and then decreased gradually on the 1st day after operation. The level of IL-25 in normal saline control group was higher than that in other groups but there was no significant difference (p0.05). On the 4th day after operation, the level of IL-25 in normal saline control group was lower than that in other groups, but there was no significant difference (p0.05). There was no significant difference in average (P0.05).
(4) The results of IL-17F test showed that the levels of IL-17F in peripheral blood of each experimental group were significantly higher than those before operation, and reached the highest level at 14 days after operation, which was significantly different from those before operation (p0.05), and then gradually decreased; the levels of IL-17F in the control group were higher than those in other groups, and there was a statistically significant difference (p0.05); The levels of IL-17F in the combined group were lower than those in the CsA low-dose group and the rhIL-10 low-dose group (p0.05). The levels of IL-17F in the rhIL-10 low-dose group were higher than those in the CsA low-dose group on the first day after operation, and the levels of IL-17F in the rhIL-10 high-dose group were higher than those in the CsA high-dose group (p0.05). At other times, there was no significant difference in the levels of IL-17 between rhIL-10 low-dose group and CSA low-dose group, rhIL-10 high-dose group and CSA high-dose group (p0.05).
CONCLUSIONS: (1) IL-17 family cytokines are significantly elevated after skin transplantation, suggesting that IL-17 family cytokines may be involved in the rejection of skin transplantation by secreting a large number of inflammatory factors to promote the secretion of inflammatory factors; (2) The changes of IL-17 family cytokines in serum may be used as a diagnostic tool for transplant rejection. Monitoring indicators: (3) inhibition of rhIL-10 on IL-17A, IL-27 and IL-17F has synergistic effect with CsA.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392.4
本文编号：2207742
[Abstract]:Objective: To observe the changes of IL-17 family factors in serum of rabbits after skin transplantation with rhIL-10, and explore the role of IL-17 family factors in the anti-rejection mechanism of rhIL-10, so as to provide experimental basis for the development and application of rhIL-10.
Methods: (1) Skin transplantation rabbits were divided into 6 groups: (1) rhIL-10 low-dose group; (2) rhIL-10 high-dose group; (3) CsA low-dose group; (4) CsA high-dose group; (6) rhIL-10 + CsA low-dose group; (3) saline group.
(2) medication regimen: 1 days before the operation, each rabbit was injected intramuscularly for 10 consecutive days.
(3) Peripheral blood samples were taken from each group one day before operation and 1,4,7,14,21,28 days after operation. Serum levels of IL-17 family cytokines were detected by enzyme-linked immunosorbent assay.
Results: (1) The results of IL-17A test showed that the levels of IL-17A in peripheral serum of rabbits in each experimental group were significantly higher than those before operation, and reached the highest level at 14 days after operation, which was statistically significant (p0.05), and then gradually decreased. The levels of IL-17A in the saline control group were higher than those in other groups, and the difference was statistically significant (p0.05). The level of IL-17A in the dosage group was lower than that in the low dose group of rhIL-10, and the difference was statistically significant (p0.05); the level of IL-17A in the combined group was lower than that in the low dose group of CsA and the low dose group of rhIL-10 was lower and statistically significant (p0.05); the level of IL-17A in the low dose group of rhIL-10 was higher than that in the low dose group of CsA on the first day after operation, and the difference was statistically significant (p0.05). There was no significant difference in IL-17A levels between rhIL-10 low dose group and CSA low dose group at other time points (p0.05).
(2) The results of IL-27 detection showed that the levels of IL-27 in peripheral serum of rabbits in each experimental group were significantly higher than those before operation, and reached the highest level at 14 days after operation, which was significantly different from those before operation (p0.05), and then gradually decreased; the levels of IL-27 in the control group were higher than those in other groups, and there was a statistically significant difference (p0.05). The level of IL-27 in the combined group was lower than that in the low-dose group of CsA and the low-dose group of rhIL-10 (p0.05). The level of IL-27 in the low-dose group of rhIL-10 was higher than that in the low-dose group of CsA at 1,4,7 days after operation, and the difference was statistically significant (p0.05). There was no significant difference between the levels of IL-27 and CSA low-dose group (p0.05), while there was no significant difference between rhIL-10 high-dose group and CSA high-dose group (p0.05).
(3) The results of IL-25 detection showed that the levels of IL-25 in peripheral serum of rabbits in each experimental group were significantly higher than those before operation, and the levels of IL-25 in normal saline control group reached the highest level on the 4th day after operation, while those in other experimental groups reached the highest level on the 14th day after operation, which was significantly different from those before operation (p0.05), and then decreased gradually on the 1st day after operation. The level of IL-25 in normal saline control group was higher than that in other groups but there was no significant difference (p0.05). On the 4th day after operation, the level of IL-25 in normal saline control group was lower than that in other groups, but there was no significant difference (p0.05). There was no significant difference in average (P0.05).
(4) The results of IL-17F test showed that the levels of IL-17F in peripheral blood of each experimental group were significantly higher than those before operation, and reached the highest level at 14 days after operation, which was significantly different from those before operation (p0.05), and then gradually decreased; the levels of IL-17F in the control group were higher than those in other groups, and there was a statistically significant difference (p0.05); The levels of IL-17F in the combined group were lower than those in the CsA low-dose group and the rhIL-10 low-dose group (p0.05). The levels of IL-17F in the rhIL-10 low-dose group were higher than those in the CsA low-dose group on the first day after operation, and the levels of IL-17F in the rhIL-10 high-dose group were higher than those in the CsA high-dose group (p0.05). At other times, there was no significant difference in the levels of IL-17 between rhIL-10 low-dose group and CSA low-dose group, rhIL-10 high-dose group and CSA high-dose group (p0.05).
CONCLUSIONS: (1) IL-17 family cytokines are significantly elevated after skin transplantation, suggesting that IL-17 family cytokines may be involved in the rejection of skin transplantation by secreting a large number of inflammatory factors to promote the secretion of inflammatory factors; (2) The changes of IL-17 family cytokines in serum may be used as a diagnostic tool for transplant rejection. Monitoring indicators: (3) inhibition of rhIL-10 on IL-17A, IL-27 and IL-17F has synergistic effect with CsA.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392.4
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