活血通腑方对大鼠术后腹腔粘连巨噬细胞细胞因子的影响
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摘要
目的探讨活血通腑方对大鼠术后腹腔粘连部位白细胞介素(IL)-6、肿瘤坏死因子(TNF)-α、精氨酸酶(Arg)-1 m RNA及IL-1、IL-6、TNF-α、IL-4、IL-10、IL-13等细胞因子的影响。方法雄性SD大鼠40只采用拙刀法建立腹腔粘连模型,分为模型组、四磨汤组、氟伐他汀组、活血通腑方组各10只,另10只为假手术组。通过粘连评分评价腹腔粘连水平,实时荧光定量聚合酶链反应(RT-PCR)检测IL-6、TNF-α、Arg-1 m RNA表达水平及酶联免疫吸附试验检测IL-1、IL-6、TNF-α、IL-4、IL-10、IL-13细胞因子含量。结果与模型组相比,四磨汤组、氟伐他汀组及活血通腑方组的粘连评分均有不同程度的减轻(P<0. 05,P<0. 01),其中活血通腑方组和四靡汤组粘连评分降低显著(P<0. 01);显著降低IL-6、TNF-α、Arg-1 m RNA表达水平及减少IL-1、IL-6、TNF-α细胞因子含量(P<0. 05);显著提高IL-4、IL-10、IL-13细胞因子含量(P<0. 05)。结论活血通腑方对大鼠术后腹腔粘连具有抑制作用,可能是通过调控巨噬细胞含量达到防治术后腹腔粘连的作用。
Objective To investigate the effect of Huoxue Tongfu decoction on IL-6,TNF-α,Arg-1 m RNA,IL-1,IL-6,TNF-α,IL-4,IL-10 and other cytokines in macrophages of postoperative intra-abdominal adhesion sites. Methods Adhesion scores were used to evaluate the level of intra-abdominal adhesions. RT-PCR was used to detect the expression levels of IL-6,TNF-α and Arg-1 m RNA. The levels of IL-1,IL-6,TNF-α,IL-4,IL-10 and IL-13 were detected by enzyme-linked immunosorbent assay. Results Huoxue Tongfu decoction could significantly reduce the score of abdominal subarachnoid adhesion. Compared with the model group,Huoxue Tongfu decoction could reduce the expressions of IL-6,TNF-α,Arg-1 m RNA and IL-1,IL-6,TNF-α cytokine content; increase IL-4,IL-10,IL-13 cytokine content.Conclusions Huoxue Tongfu decoction has inhibitory effect on abdominal adhesion in rats after operation. It may be effective by regulating the content of macrophages to prevent and cure postoperative abdominal adhesion.
Objective To investigate the effect of Huoxue Tongfu decoction on IL-6,TNF-α,Arg-1 m RNA,IL-1,IL-6,TNF-α,IL-4,IL-10 and other cytokines in macrophages of postoperative intra-abdominal adhesion sites. Methods Adhesion scores were used to evaluate the level of intra-abdominal adhesions. RT-PCR was used to detect the expression levels of IL-6,TNF-α and Arg-1 m RNA. The levels of IL-1,IL-6,TNF-α,IL-4,IL-10 and IL-13 were detected by enzyme-linked immunosorbent assay. Results Huoxue Tongfu decoction could significantly reduce the score of abdominal subarachnoid adhesion. Compared with the model group,Huoxue Tongfu decoction could reduce the expressions of IL-6,TNF-α,Arg-1 m RNA and IL-1,IL-6,TNF-α cytokine content; increase IL-4,IL-10,IL-13 cytokine content.Conclusions Huoxue Tongfu decoction has inhibitory effect on abdominal adhesion in rats after operation. It may be effective by regulating the content of macrophages to prevent and cure postoperative abdominal adhesion.
引文
1曾莉.腹腔粘连研究进展[J].南京中医药大学学报,2004;20(2):126-8.
2 Wei G,Chen X,Wang G,et al.Effect of resveratrol on the prevention of intra-abdominal adhesion formation in a rat model[J].Cell Physiol Biochem,2016;39(1):33-46.
3 Jin X,Ren S,Macarak E,et al.Pathobiological mechanisms of peritoneal adhesions:the mesenchymal transition of rat peritoneal mesothelial cells induced by TGF-beta1 and IL-6 requires activation of Erk1/2and Smad2 linker region phosphorylation[J].Matrix Biol,2016;4(51):55-64.
4 Arslan E,Irkorucu O,Sozutek A,et al.The potential efficacy of Survanta(r)and Seprafilm(r)on preventing intra-abdominal adhesions in rats[J].Acta Cir Bras,2016;31(6):389-95.
5赵义江,张国志,王长友,等.腹腔粘连的研究现状分析[J].华北煤炭医学院学报,2011;13(6):777-9.
6颜帅,曾莉,李文林,等.从腹腔粘连论中医“结”病机学说[J].中医杂志,2014;55(19):1641-4.
7曾莉,钱海华,赵群男,等.活血通腑方治疗术后粘连性肠梗阻56例临床观察[J].南京中医药大学学报,2010;26(3):178-80.
8颜帅,杨斓,曾莉,等.活血通腑方优化方防治大鼠术后腹腔粘连的最佳有效剂量[J].中国实验方剂学杂志,2014;20(9):166-70.
9陆二梅,李益星,白杨,等.巨噬细胞极化的研究进展[J].现代生物医学进展,2016;16(32):6397-400.
10宮甜甜,黄少刚,张玥,等.巨噬细胞的极化及功能调控[J].解剖学报,2017;48(1):106-10.
11颜帅.活血通腑方干预CD40/CD40L共刺激通路抗腹腔粘连的实验研究[D].南京:南京中医药大学,2015.
12 Phillips RK,Dudley HA.The effect of tetracycline lavage and trauma on visceral and parietal peritoneal ultrastructure and adhesion formation[J].Br J Surg,1984;71(7):537-9.
13李爱科,魏凤玲,原国强,等.通腑、活血中药防治术后肠粘连的实验研究[J].北京中医药大学学报,2001;24(5):23-5.
14陈木水.活血祛瘀法在钝性腹部损伤中的应用[J].中国中西医结合急救杂志,1996;3(2):65-6.
15曾莉,徐庆,夏卫军,等.活血通腑方治疗腹腔粘连处方优化的正交T值法研究[J].中国中医急症,2008;17(3):370-1.
16曾莉,徐庆,陈卫平,等.活血通腑方优选方对TNF-α诱导下腹膜间皮细胞TGF-β1、FN、CTGF蛋白含量的影响[J].南京中医药大学学报,2008;24(5):324-6,364.
17曾莉,翟亚春,夏国守,等.活血通腑方有效剂量筛选实验研究[J].南京中医药大学学报,2004;20(5):309-11.
18曾莉,颜帅,李文林,等.活血通腑方对术后腹腔粘连大鼠单核细胞趋化蛋白-1及CD40/CD40L的影响[J].中成药,2015;37(10):2101-8.
19 Zhang H,Song Y,Li Z,et al.Evaluation of breviscapine on prevention of experimentally induced abdominal adhesions in rats[J].Am JSurg,2016;211(6):1143-52.
20沈晓婷,成臣,万贵平.巨噬细胞及其相关因子在子宫内膜异位症中的表达及作用[J].浙江临床医学,2015;17(2):313-5.
21 Sindrilaru A,Peters T,Wieschalka S,et al.An unrestrained proinflammatory M1 macrophage population induced by iron impairs wound healing in humans and mice[J].J Clin Invest,2011;121(3):985-97.
22 Benoit M,Desnues B,Mege JL.Macrophage polarization in bacterial infections[J].J Immunol,2008;181(6):3733-9.
23田维毅,葛金文.M1/M2型巨噬细胞极化对动脉粥样硬化炎症影响的研究进展[J].贵阳中医学院学报,2014;36(6):18-21.
24 Yang Z,Grinchuk V,Urban JF Jr,et al.Macrophages as IL-25/IL-33-responsive cells play an important role in the induction of type 2immunity[J].PLoS One,2013;8(3):e59441.
2 Wei G,Chen X,Wang G,et al.Effect of resveratrol on the prevention of intra-abdominal adhesion formation in a rat model[J].Cell Physiol Biochem,2016;39(1):33-46.
3 Jin X,Ren S,Macarak E,et al.Pathobiological mechanisms of peritoneal adhesions:the mesenchymal transition of rat peritoneal mesothelial cells induced by TGF-beta1 and IL-6 requires activation of Erk1/2and Smad2 linker region phosphorylation[J].Matrix Biol,2016;4(51):55-64.
4 Arslan E,Irkorucu O,Sozutek A,et al.The potential efficacy of Survanta(r)and Seprafilm(r)on preventing intra-abdominal adhesions in rats[J].Acta Cir Bras,2016;31(6):389-95.
5赵义江,张国志,王长友,等.腹腔粘连的研究现状分析[J].华北煤炭医学院学报,2011;13(6):777-9.
6颜帅,曾莉,李文林,等.从腹腔粘连论中医“结”病机学说[J].中医杂志,2014;55(19):1641-4.
7曾莉,钱海华,赵群男,等.活血通腑方治疗术后粘连性肠梗阻56例临床观察[J].南京中医药大学学报,2010;26(3):178-80.
8颜帅,杨斓,曾莉,等.活血通腑方优化方防治大鼠术后腹腔粘连的最佳有效剂量[J].中国实验方剂学杂志,2014;20(9):166-70.
9陆二梅,李益星,白杨,等.巨噬细胞极化的研究进展[J].现代生物医学进展,2016;16(32):6397-400.
10宮甜甜,黄少刚,张玥,等.巨噬细胞的极化及功能调控[J].解剖学报,2017;48(1):106-10.
11颜帅.活血通腑方干预CD40/CD40L共刺激通路抗腹腔粘连的实验研究[D].南京:南京中医药大学,2015.
12 Phillips RK,Dudley HA.The effect of tetracycline lavage and trauma on visceral and parietal peritoneal ultrastructure and adhesion formation[J].Br J Surg,1984;71(7):537-9.
13李爱科,魏凤玲,原国强,等.通腑、活血中药防治术后肠粘连的实验研究[J].北京中医药大学学报,2001;24(5):23-5.
14陈木水.活血祛瘀法在钝性腹部损伤中的应用[J].中国中西医结合急救杂志,1996;3(2):65-6.
15曾莉,徐庆,夏卫军,等.活血通腑方治疗腹腔粘连处方优化的正交T值法研究[J].中国中医急症,2008;17(3):370-1.
16曾莉,徐庆,陈卫平,等.活血通腑方优选方对TNF-α诱导下腹膜间皮细胞TGF-β1、FN、CTGF蛋白含量的影响[J].南京中医药大学学报,2008;24(5):324-6,364.
17曾莉,翟亚春,夏国守,等.活血通腑方有效剂量筛选实验研究[J].南京中医药大学学报,2004;20(5):309-11.
18曾莉,颜帅,李文林,等.活血通腑方对术后腹腔粘连大鼠单核细胞趋化蛋白-1及CD40/CD40L的影响[J].中成药,2015;37(10):2101-8.
19 Zhang H,Song Y,Li Z,et al.Evaluation of breviscapine on prevention of experimentally induced abdominal adhesions in rats[J].Am JSurg,2016;211(6):1143-52.
20沈晓婷,成臣,万贵平.巨噬细胞及其相关因子在子宫内膜异位症中的表达及作用[J].浙江临床医学,2015;17(2):313-5.
21 Sindrilaru A,Peters T,Wieschalka S,et al.An unrestrained proinflammatory M1 macrophage population induced by iron impairs wound healing in humans and mice[J].J Clin Invest,2011;121(3):985-97.
22 Benoit M,Desnues B,Mege JL.Macrophage polarization in bacterial infections[J].J Immunol,2008;181(6):3733-9.
23田维毅,葛金文.M1/M2型巨噬细胞极化对动脉粥样硬化炎症影响的研究进展[J].贵阳中医学院学报,2014;36(6):18-21.
24 Yang Z,Grinchuk V,Urban JF Jr,et al.Macrophages as IL-25/IL-33-responsive cells play an important role in the induction of type 2immunity[J].PLoS One,2013;8(3):e59441.