异体红细胞、白细胞对IL-8、IL-10、IL-12调控的实验研究

发布时间：2018-05-03 09:06

本文选题：输血相关免疫调节 + 异体红细胞　；参考：《第二军医大学》2012年硕士论文

【摘要】：（一）目的运用郭氏“系统血液快速固有免疫反应分离实验体系”体外观察异体红细胞对IL-8、IL-10、IL-12分泌的影响。（二）方法健康成人同型新鲜抗凝全血，常规分离制备血浆、红细胞和白细胞，以自体血浆作为反应基质建立系统血液分离实验体系，考察异体红细胞对IL-8、IL-10、IL-12分泌的影响，设立自体红细胞作为对照。ELISA法测定上清液中的细胞因子IL-8、IL-10、IL-12，并探讨其改变可能存在的意义。（三）结果 1、血浆IL-8浓度比较 ⑴异体红细胞生理盐水体系组[28.89179±2.674565pg/ml]与异体红细胞人大肠埃希杆菌体系组[119.05455±31.056229pg/ml）比较，其均值差值[-90.1628±59.37619pg/ml],（p=0.0050.05），IL-8分泌量两组之间有着显著的差别。 ⑵自体红细胞生理盐水体系组[29.05148±2.972132pg/ml]与自体红细胞大肠埃希杆菌体系组[132.63303±35.072837pg/ml]比较，其均值差值[-103.5816±59.37619pg/ml],（p=0.0000.05），IL-8分泌量两组之间有着显著的差别。 ⑶异体红细胞生理盐水体系组与自体红细胞生理盐水体系组相比较，其均值差值[-0.1599±59.37619pg/ml]，IL-8分泌量两组之间没有明显的差别（p=0.8190.05）。 ⑷异体红细胞人大肠埃希杆菌体系组与自体红细胞大肠埃希杆菌体系组相比较，其均值差值[-13.5785±59.37619]，IL-8分泌量两组之间没有明显的差别（p=0.9970.05）。 2、血浆IL-10浓度比较 ⑴异体红细胞生理盐水体系组[27.90894±5.445463pg/ml]与异体红细胞人大肠埃希杆菌体系组[22.28497±4.010929pg/ml]比较，其均值差值[5.6240±5.37453pg/ml]，（p=0.2960.05） IL-10分泌量两组之间没有显著的差别。 ⑵自体红细胞生理盐水体系组[24.15185±3.748592pg/ml]与自体红细胞大肠埃希杆菌体系组[27.58494±4.740832pg/ml]比较，其均值差值[-3.4331±5.37453pg/ml（p=0.5240.05），IL-10分泌量两组之间没有显著的差别。 ⑶异体红细胞生理盐水体系组与自体红细胞生理盐水体系组相比较，其均值差值[3.7571±5.37453pg/ml] IL-10分泌量两组之间没有显著的差别（p=0.3250.05）。 ⑷异体红细胞人大肠埃希杆菌体系组与自体红细胞大肠埃希杆菌体系组相比较，其均值差值[-5.2999±5.37453pg/ml]，IL-10分泌量两组之间没有明显的差别（p=0.4850.05）。 3、IL-12分泌量的测定值 ⑴异体红细胞生理盐水体系组[25.80663±3.872463pg/ml]与异体红细胞人大肠埃希杆菌体系组[22.97730±2.298994pg/ml]比较，其均值差值[2.8289±4.48095pg/ml]，（p=0.5280.05），IL-12分泌量两组之间没有显著的差别。 ⑵自体红细胞生理盐水体系组[37.74982±5.783372pg/ml]与自体红细胞大肠埃希杆菌体系组[33.32391±2.982255pg/ml]比较，其均值差值[4.4259±4.48095pg/ml]，（p=0.3240.05），IL-12分泌量两组之间没有显著的差别。 ⑶异体红细胞生理盐水体系组与自体红细胞生理盐水体系组相比较，其均值差值[-11.9436±4.48095pg/ml]，IL-12分泌量两组之间有着显著的差别（p=0.0080.05）。 ⑷异体红细胞人大肠埃希杆菌体系组与自体红细胞大肠埃希杆菌体系组相比较，其均值[-10.3466±4.48095pg/ml]，（p=0.0220.05），IL-12分泌量两组之间有着显差别。（四）结论异体红细胞实验组相对于自体红细胞对照组IL-8、IL-10的分泌量没有显著的不同，表明异体红细胞对单核-巨噬细胞、Th2等细胞分泌细胞因子IL-8、IL-10的功能没有明显的调节作用；异体红细胞实验组分泌IL-12的量显著低于自体红细胞对照组，表明异体红细胞对DC细胞、巨噬细胞等细胞分泌细胞因子IL-12的能力有着一定程度的负向调控作用。（一）目的运用郭氏“系统血液快速固有免疫反应分离实验体系”体外观察异体白细胞对IL-8、IL-10、IL-12分泌的影响。（二）方法健康成人同型新鲜抗凝全血，常规分离制备血浆、红细胞和白细胞，以自体血浆作为反应基质建立系统血液分离实验体系，考察异体白细胞对IL-8、IL-10、IL-12分泌的影响，设立自体白细胞作为对照。ELISA法测定上清液中的细胞因子IL-8、IL-10、IL-12，并探讨其改变可能存在的意义。（三）结果 1、血浆IL-8浓度比较 ⑴异体白细胞生理盐水体系[31.12373±2.828597pg/ml]组与异体白细胞人大肠埃希杆菌体系组[199.75970±62.743079pg/ml]比较，其均值差值[-168.6360±59.37619pg/ml]，（p=0.0050.05），IL-8分泌量两组之间有着显著的差别。 ⑵自体白细胞生理盐水体系[48.29476±6.511376pg/ml]组与自体白细胞大肠埃希杆菌体系组[465.92697±111.354713pg/ml]比较，其均值差值[-417.6322±59.37619pg/ml],（p=0.0000.05）,IL-8分泌量两组之间有着显著的差别。 ⑶异体白细胞生理盐水体系组与自体白细胞生理盐水体系组相比较，其均值差值[-17.1710±59.37619pg/ml]，IL-8分泌量两组之间没有明显的差别（p=0.7730.05）。 ⑷异体白细胞人大肠埃希杆菌体系组与自体白细胞大肠埃希杆菌体系组相比较，其均值差值[-266.1673±59.37619pg/ml]，IL-8分泌量两组之间没有明显的差别（p=0.0000.05）。 2、血浆IL-10浓度比较 ⑴异体白细胞生理盐水体系组[21.37236±3.307657pg/ml]与异体白细胞人大肠埃希杆菌体系组[16.93582±2.346794pg/ml]比较，其均值差值[4.4365±5.37453pg/ml]，（p=0.4100.05）， IL-10分泌量两组之间没有显著的差别。 ⑵自体白细胞生理盐水体系组[47.65618±7.432838pg/ml]与自体白细胞大肠埃希杆菌体系组[46.16091±9.180075pg/ml]比较，其均值差值[1.4953±5.37453pg/ml],（p=0.7810.05），IL-10分泌量两组之间没有显著的差别。 ⑶异体白细胞生理盐水体系组与自体白细胞生理盐水体系组相比较，其均值差值[-26.2838±5.37453pg/ml]， IL-10分泌量两组之间有显著的差别（p=0.0000.05）。 ⑷异体白细胞人大肠埃希杆菌体系组与自体白细胞大肠埃希杆菌体系组相比较，其均值差值[-29.2251±5.37453pg/ml]，IL-10分泌量两组之间有显著的差别（p=0.0000.05）。 3、IL-12分泌量的测定值 ⑴异体白细胞生理盐水体系组[47.30961±8.11704pg/ml]与异体白细胞人大肠埃希杆菌体系组[39.69006±6.517480pg/ml]比较，其均值差值[7.6402±4.48095pg/ml]（p=0.6650.05），IL-12分泌量两组之间没有明显的差别。 ⑵自体白细胞生理盐水体系组[39.95673±4.607648pg/ml]与自体白细胞大肠埃希杆菌体系组[29.00088±3.838735pg/ml]比较，其均值差值[10.9558±4.48095pg/ml],（p=0.0150.05），IL-12分泌量两组之间有显著的差别。 ⑶异体白细胞生理盐水体系组与自体白细胞生理盐水体系组相比较，其均值差值[7.3529±4.48095pg/ml]，IL-12分泌量两组之间没有明显的差别（p=0.1020.05）。 ⑷异体白细胞人大肠埃希杆菌体系组与自体白细胞大肠埃希杆菌体系组相比较[10.6892±4.48095pg/ml]，，（p=0.0180.05），IL-12分泌量两组之间有着显差别。（四）结论异体白细胞实验组相对于自体白细胞对照组IL-8、IL-10的分泌显著降低，说明异体白细胞对单核-巨噬细胞及Th2细胞等细胞分泌细胞因子IL-8、IL-10有着明显的负向调节作用；异体白细胞实验组与自体白细胞对照组相比较IL-8与IL-10的分泌量显著下降且呈明显的正相关性，至少表明这两种细胞因子之间相互调节，以达成细胞因子网络的稳态平衡；异体白细胞实验组相对于自体白细胞对照组IL-12的量显著降低，表明异体白细胞对DC细胞、巨噬细胞等细胞分泌细胞因子IL-12的能力有着一定程度的负向调控作用。
[Abstract]:( i ) Purpose

The effects of allogeneic red blood cells on the secretion of IL - 8 , IL - 10 and IL - 12 were observed in vitro by using the " system of rapid innate immune reaction in blood " of Guo ' s system .

( II ) Methodology

The effects of allogeneic red blood cells on the secretion of IL - 8 , IL - 10 and IL - 12 were determined by ELISA . The levels of IL - 8 , IL - 10 , IL - 12 in supernatant were determined by ELISA .

( III ) Results

1 . Comparison of plasma IL - 8 concentration

( 1 ) There was a significant difference between the two groups : ( 1.89179 卤 2.674565pg / ml ) of allogeneic red blood cell physiological saline group compared with ( 119.05455 卤 31.056229pg / ml ) .

( 2 ) There was a significant difference between the two groups of autologous red blood cell physiological saline system group ( 29.05148 卤 2.972132pg / ml ) and autologous red blood cell large intestine Escherichia coli system group ( 132.63303 卤 35.072837pg / ml ) .

There was no significant difference between the two groups ( p = 0.819 . 05 ) .

( 4 ) There was no significant difference between the two groups ( p = 0.9970 . 05 ) .

2 . Comparison of plasma IL - 10 concentration

The mean difference was 5.6240 卤 5.37453pg / ml , and there was no significant difference between the two groups ( p = 0 . 2960 . 05 ) .

( 2 ) There was no significant difference between IL - 10 secretion and IL - 10 secretion in autologous red cell physiological saline system group ( 24.15185 卤 3.748592pg / ml ) and autologous red blood cell large intestine Escherichia coli system group ( 27.58494 卤 4.740832pg / ml ) .

The mean difference was 3.7571 卤 5.37453pg / ml and there was no significant difference between the two groups ( p = 0.3250.05 ) .

( 4 ) There was no significant difference between the two groups ( p = 0.4850.05 ) .

3 . Measured value of IL - 12 secretion

The mean difference was 2.8289 卤 4.48095pg / ml , ( p = 0.5280.05 ) and there was no significant difference between the two groups of IL - 12 secretion .

( 2 ) There was no significant difference between the two groups of autologous red blood cell physiological saline system group ( 37.74982 卤 5.783372pg / ml ) and autologous red blood cell large intestine Escherichia coli system group ( 33.323 91 卤 2.982255pg / ml ) , and the mean difference was 4.4259 卤 4.48095pg / ml respectively , ( p = 0.3240.05 ) and IL - 12 secretion .

( 3 ) There was a significant difference between the two groups ( p = 0.0080.05 ) .

( 4 ) Compared with the autologous red cell E . coli system group , the average value of E . coli was 10.3466 卤 4.48095pg / ml , ( p = 0.0220 . 05 ) , and there was a significant difference between the two groups of IL - 12 secretion .

( IV ) Conclusion

There was no significant difference in the secretion of IL - 8 and IL - 10 from the experimental group , indicating that allogeneic red blood cells had no obvious regulation effect on the secretion of cytokines IL - 8 and IL - 10 by mononuclear - macrophage , Th2 , etc .
The amount of IL - 12 secreted by allogeneic red blood cells in experimental group was significantly lower than that of autologous red blood cell control group , indicating that the ability of allogeneic red blood cells to secrete cytokines IL - 12 in DC cells , macrophages and other cells has a certain degree of negative regulation .

( i ) Purpose

The effect of allogeneic white blood cell on IL - 8 , IL - 10 and IL - 12 secretion was observed in vitro by using the " system of rapid innate immune reaction in blood " of Guo ' s system .

( II ) Methodology

The effects of allogeneic white blood cells on the secretion of IL - 8 , IL - 10 and IL - 12 were determined by ELISA . The levels of IL - 8 , IL - 10 , IL - 12 in supernatant were determined by ELISA .

( III ) Results

1 . Comparison of plasma IL - 8 concentration

( 1 ) There was a significant difference between IL - 8 secretion and IL - 8 secretion between the two groups . The mean difference of IL - 8.6360 卤 59.37619pg / ml , ( p = 0.0050.05 ) and IL - 8 secretion were significantly different between the two groups .

( 2 ) The mean value difference of autologous white blood cell physiological saline system ( 48.29476 卤 6.511376pg / ml ) was significantly different from that of autologous white cell E . coli system group ( 465.92697 卤 111.354713pg / ml ) , and the mean difference of IL - 417.6322 卤 59.37619pg / ml ) , ( p = 0.00000.05 ) and IL - 8 secretion .

There was no significant difference between the two groups ( p = 0.777 . 05 ) .

( 4 ) There was no significant difference between the two groups ( p = 0.0000.05 ) .

2 . Comparison of plasma IL - 10 concentration

The mean difference was 4.4365 卤 5.37453pg / ml , ( p = 0.41 . 05 ) , and there was no significant difference between the two groups .

The mean difference was 1.4953 卤 5.37453pg / ml , ( p = 0.781 . 05 ) and there was no significant difference between the two groups .

( 3 ) Compared with the normal saline system group , the mean difference of IL - 10 secretion was significantly different between the two groups ( p = 0.0000 . 05 ) .

( 4 ) There was a significant difference between the two groups ( p = 0.0000 . 05 ) .

3 . Measured value of IL - 12 secretion

The mean difference was 7.6402 卤 4.48095pg / ml ( p = 0.665 . 05 ) and there was no significant difference between the two groups of IL - 12 secretion .

The mean difference was 10.9558 卤 4.48095pg / ml , ( p = 0.0150.05 ) , and there was a significant difference between the two groups of IL - 12 secretion .

The mean difference was 7.3529 卤 4.48095pg / ml , and there was no significant difference between the two groups ( p = 0.1020 . 05 ) .

( 4 ) There was a significant difference between the two groups ( 10.6892 卤 4.48095pg / ml ) , ( p = 0.0180 . 05 ) and IL - 12 secretion .

( IV ) Conclusion

The secretion of IL - 8 and IL - 10 decreased significantly in the experimental group , and the IL - 8 and IL - 10 secretion of allogeneic white blood cells were negatively regulated by IL - 8 and IL - 10 .
The levels of IL - 8 and IL - 10 decreased and positively correlated with IL - 8 and IL - 10 in the experimental group .
Compared with the control group , the amount of IL - 12 decreased significantly in the experimental group , indicating that the ability of allogeneic white blood cells to secrete cytokines IL - 12 in DC cells , macrophages and other cells has a certain degree of negative regulation .

【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

【参考文献】