腹腔神经丛阻滞对大鼠肝部分切除术后应激反应和炎症因子的影响及其机制

doi:10.13481/j.1671-587X.20210221

摘要/Abstract

摘要： 目的

探讨腹腔神经丛阻滞对大鼠肝部分切除术后应激反应及免疫炎性的影响，并阐明其作用机制。

方法

取40只大鼠，随机选取10只作为正常组，10只作为阻滞剂对照剂组，20只制备大鼠肝部分切除术模型。取造模成功的16只大鼠，随机分为模型组和模型+阻滞剂组，每组8只。模型+阻滞剂组大鼠于术毕关腹前给予腹腔神经丛双侧注射0.5%利多卡因，阻滞剂对照组大鼠不制备模型，给予腹腔神经丛双侧注射0.5%利多卡因，正常组与模型组大鼠给予腹腔神经丛双侧注射等量生理盐水。给药12 h后，酶联免疫吸附试验（ELISA）法检测各组大鼠血清中皮质酮（CORT）、促肾上腺皮质激素（ACTH）、去甲肾上腺素（NE）、肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）和白细胞介素6（IL-6）水平，实时荧光定量聚合酶链式反应（RT-qPCR）和Western blotting法检测各组大鼠外周血单个核细胞中糖皮质激素受体（GR）、Toll样受体4（TLR4）mRNA和蛋白表达水平。

结果

与正常组和阻滞剂对照组比较，模型组和模型+阻滞剂组大鼠血清中CORT、ACTH和NE水平升高（P<0.05）；与模型组比较，模型+阻滞剂组大鼠血清中CORT、ACTH和NE水平降低（P<0.05）。与正常组和阻滞剂对照组比较，模型组和模型+阻滞剂组大鼠血清中TNF-α、IL-1β和IL-6水平升高（P<0.05）；与模型组比较，模型+阻滞剂组大鼠血清中TNF-α、IL-1β和IL-6水平降低（P<0.05）。与正常组和阻滞剂对照组比较，模型组和模型+阻滞剂组大鼠外周血单个核细胞中GR mRNA及蛋白表达水平升高（P<0.05），TLR4 mRNA和蛋白表达水平降低（P<0.05）；与模型组比较，模型+阻滞剂组大鼠外周血单个核细胞中GR mRNA和蛋白表达水平升高（P<0.05），TLR4 mRNA和蛋白表达水平降低（P<0.05）。正常组和阻滞剂对照组大鼠血清中CORT、ACTH、NE、TNF-α、IL-1β、IL-6水平及外周血单个核细胞中GR、TLR4 mRNA和蛋白表达水平比较，差异无统计学意义（P>0.05）。

结论

腹腔神经丛阻滞能减轻肝部分切除术后大鼠的应激反应及免疫炎性反应，其机制可能与上调GR蛋白表达和下调TLR4蛋白表达有关。

关键词: 肝部分切除术, 腹腔神经丛阻滞, 应激反应, 免疫炎性, 糖皮质激素受体, Toll样受体4

Abstract: Objective

To discuss the effect of celiac plexus block on the stress response and immune inflammation of the rats after partial hepatectomy， and to clarify its mechanism.

Methods

Forty rats were selected， ten rats were randomly selected as normal group， ten rats were used as blocker control group， and the other 20 rats were used to construct partial hepatectomy models. Sixteen modeling rats were randomly divided into model group and model + blocker group， and there were eight rats in each group. The rats in model + blocker group were injected with 0.5% lidocaine on both sides of the celiac plexus before abdominal closure， and the rat models were not prepared in blocker control group and the rats were injected with 0.5% lidocaine on both sides of the celiac plexus； the rats in normal group and model group were injected with the same amount of normal saline. At 12 h after of administration， the levels of cocorticoid （CORT），adrenocorticotropic hormone （ACTH）， noradrenaline （NE）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in serum of the rats in various groups were measured by ELISA method；Real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to detect the expression levels of glucocorticoid receptor （GR） and Toll-like receptor 4 （TLR4） mRNA and proteins in the peripheral blood mononuclear cells of the rats in various groups.

Results

Compared with normal group and blocker control group， the levels of CORT， ACTH and NE in serum of the rats in model group and model + blocker group were increased （P<0.05）； compared with model group， the levels of CORT， ACTH， and NE in serum of the rats model + blocker group were decreased （P<0.05）. Compared with normal group and blocker control group， the levels of TNF-α， IL-1β and IL-6 in serum of the rats in model group and model + blocker group were increased （P<0.05）； compared with model group， the levels of TNF-α， IL-1β， and IL-6 in serum of the rats in model + blocker group were decreased （P<0.05）. Compared with normal group and blocker control group， the expression levels of GR mRNA and protein in the peripheral blood mononuclear cells of the rats in model group and model+blocker group were increased（P<0.05）， while the expression levels of TLR4 mRNA and protein were decreased （P<0.05）； compared with model group， the expression levels of GR mRNA and protein in the peripheral blood mononuclear cells of the rats in model + blocker group were increased（P<0.05）， while the expression levels of TLR4 mRNA and protein were decreased （P<0.05）. There were no significant differences in the serum levels of CORT， ACTH， NE， TNF-α， IL-1β， and IL-6， and the expression levels of GR and TLR4 mRNA and proteins in peripheral blood mononuclear cells of the rats between normal group and blocker control group （P>0.05）.

Conclusion

Celiac plexus block can reduce the stress response and immune inflammatory response of the rats after partial hepatectomy， and its mechanism may be related to up-regulating the GR protein expression and down-regulating the TLR4 protein expression.

Key words: partial hepatectomy, celiac plexus block, stress response, immunoinflammatory, glucocorticoid receptor, Toll-like receptor 4

中图分类号:

R657.3

马臻杰,马兰,贾珍. 腹腔神经丛阻滞对大鼠肝部分切除术后应激反应和炎症因子的影响及其机制[J]. 吉林大学学报(医学版), 2021, 47(2): 407-413.

Zhenjie MA,Lan MA,Zhen JIA. Effect of celiac plexus block on stress response and immune inflammation of rats after partial hepatectomy and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2021, 47(2): 407-413.

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Primer	Sequence (5'－3')
GR	Forward:TGAGATGTATATGTTGTA
GR	Reverse:ATCAGAGTGATCAGCGT
TLR4	Forward:GCATGCATAGTGTATAGT
TLR4	Reverse:TGAACTACATAATCTATG
β-actin	Forward:ATGGAGACACGTATGCT
β-actin	Reverse:TACTATCTGAGTGTACGT

Group	n	CORT	ACTH	NE
Normal	10	81.56±9.15	69.14±7.45	215.32±30.25
Blocker control	10	80.49±8.95	68.95±7.11	205.14±29.32
Model	8	115.68±11.65^*△	102.44±12.33^*△	316.78±31.25^*△
Model+blocker	8	98.31±10.25^*△#	81.69±11.25^*△#	259.36±26.98^*△#
F		24.330	23.623	21.545
P		<0.01	<0.01	<0.01

Group	n	TNF-α	IL-1β	IL-6
Normal	10	64.01±7.12	16.32±2.31	19.55±2.66
Blocker control	10	66.03±7.09	18.12±2.58	21.98±3.65
Model	8	94.25±10.12^*△	55.33±6.65^*△	59.45±7.12^*△
Model+blocker	8	79.32±9.68^*△#	32.25±4.21^*△#	34.66±4.56^*△#
F		23.951	165.885	134.603
P		<0.01	<0.01	<0.01

Group	n	GR mRNA	TLR4 mRNA
Normal	10	0.98±0.11	0.31±0.04
Blocker control	10	0.97±0.13	0.34±0.05
Model	8	0.48±0.06^*△	0.88±0.11^*△
Model+blocker	8	0.71±0.09^*△#	0.59±0.07^*△#
F		46.526	125.559
P		<0.01	<0.01

Group	n	GR	TLR4
Normal	10	0.98±0.11	0.31±0.04
Blocker control	10	0.96±0.13	0.34±0.05
Model	8	0.60±0.10^*△	0.75±0.11^*△
Model+blocker	8	0.76±0.11^*△#	0.58±0.07^*△#
F		23.904	79.702
P		<0.01	<0.01