Effect of Didangtang on Inflammatory Reaction in Myocardium of DM Rats of by Modulating NF-B Pathway

CHU Quan-gen; WANG Ling; ZHANG Kai; PAN Xiao-feng

doi:10.13422/j.cnki.syfjx.2018010109

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Effect of Didangtang on Inflammatory Reaction in Myocardium of DM Rats of by Modulating NF-B Pathway

更新时间：2024-01-04

- Effect of Didangtang on Inflammatory Reaction in Myocardium of DM Rats of by Modulating NF-B Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 1, Pages: 109-113(2018)
- 作者机构：
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.2018010109
  CLC： R285.5
- Published：2018
- 稿件说明：
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CHU Quan-gen, WANG Ling, ZHANG Kai, et al. Effect of Didangtang on Inflammatory Reaction in Myocardium of DM Rats of by Modulating NF-B Pathway[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(1): 109-113.
DOI：

CHU Quan-gen, WANG Ling, ZHANG Kai, et al. Effect of Didangtang on Inflammatory Reaction in Myocardium of DM Rats of by Modulating NF-B Pathway[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(1): 109-113. DOI： 10.13422/j.cnki.syfjx.2018010109.

摘要

目的：探讨抵当汤及其蕴含的泻热化瘀通络法对因核转录因子-кB（NF-κB）通路激活而导致的糖尿病（diabetes mellitus，DM）大鼠心肌炎症反应的影响。方法：选取130只体重达到220~270 g清洁级SD健康雄性大鼠，分为正常组（10只）和造模组（120只），后者按55 mg·kg-1剂量一次性腹腔注射链脲佐菌素（streptozotocin，STZ）溶液建立DM模型。72 h后将造模成功的大鼠随机分为模型组，抵当汤高、中、低剂量组（1.08，0.72，0.54 g·kg-1·d-1），吡咯啉烷二甲基硫脲组（pyrrolidine dithiocarbamate，PDTC组，0.15 g·kg-1·d-1）以及抵当-吡咯啉烷二甲基硫脲组（DP组1.08 g·kg-1·d-1抵当汤+0.15 g·kg-1·d-1 PDTC），均采用相应药物1次/d进行灌胃干预。8周后，麻醉状态下腹主动脉取血处死动物，迅速摘取大鼠心脏，置于液氮中储存备用。以蛋白免疫印迹法（Western blot）检测心肌组织匀浆中转录NF-κB p65蛋白的表达，酶联免疫吸附法（ELISA）测定糖尿病心肌病（DCM）大鼠心肌组织匀浆肿瘤坏死因子-a（tumor necrosis factor-alpha，TNF-α）表达，免疫荧光检测心肌组织细胞黏附分子-1（intercellular cell adhesion molecule-1，ICAM-1），巨噬细胞趋化性蛋白-1（macrophage chemotaxis protein-1，MCP-1）的表达。结果：与正常组比较，模型组NF-κB p65蛋白和TNF-α表达均明显升高；且模型组胞浆内观察到被染成绿色荧光的ICAM-1，MCP-1蛋白。与模型组比较，各治疗组NF-κB p65蛋白和TNF-α表达则明显降低；组间比较发现，DP组NF-κB p65蛋白表达与正常组最接近，抵当汤高剂量组TNF-α表达则较模型组降低更加显著；各治疗组胞浆内观察到被染成绿色荧光的ICAM-1，MCP-1蛋白不同程度减少，其中DP组被染成绿色的范围最少，与正常组最为接近。结论：泻热化瘀通络之抵当汤可通过抑制NF-κB通路的激活来减轻DM模型大鼠的心肌炎症反应。

Abstract

Objective:To investigate the effect of Didangtang in purging heat

dispersing blood stasis and dredging collaterals on myocardial inflammation reaction of diabetes mellitus (DM) rats by activating nuclear factor-κB(NF-κB) pathway. Method:A total of 130 clean grade healthy male SD rats weighing between 220 g and 270 g were selected and divided into the normal group (10 rats) and the model group (120 rats). The rats of the model group were injected with 55 mg·kg-1 streptozotocin (STZ) solution to establish the DM model. After 72 hours

the rats were randomly divided into the high-dose group

the model group

the low-dose group (1.08

0.72

0.54 g·kg-1·d-1)

the pyrrolylene dimethyl thiourea group (PDTC group 0.15 g·kg-1·d-1) and the didang-pyrrolidine dimethyl thiourea group (DP group 1.08 g·kg-1·d-1+0.15 g·kg-1·d-1 PDTC)

and intervened with the corresponding drugs once a day by gavage. After 8 weeks

the animals were put to death by drawing blood from the abdominal aorta in anesthesia. And then the rats' hearts were quickly removed and stored in liquid nitrogen. The NF-кB p65 protein expression in myocardium homogenate was detected by Western blot

the expression of tumor necrosis factor-α(TNF-α) was detected by enzyme-linked immunosorbent assay(ELISA) method

and the expressions of intercellular cell adhesion molecule-1 (ICAM-1) and macrophage chemotaxis protein-1 (MCP-1) in myocardium were detected by immunofluorescence. Result:The expressions of NF-кB p65 and TNF-α of the model group were significantly higher than those in the normal group. And we observed that ICAM-1 and MCP-1 proteins were dyed as green fluorescent in cytoplasm in the model group. Compared with the model group

the expressions of NF-кB p65 protein and TNF-α in the treatment groups were significantly lower. According to the inter-group comparison

the expression of NF-кB p65 protein in the DP group was the closest to that in the normal group

and the expression of TNF-α in the high-dose Didangtang group was significantly lower than that in the model group; the treatment groups showed that cytoplasm were dyed as green fluorescent and ICAM-1 and MCP-1 proteins decreased in different degrees

and the DP group was the closest to the normal group

with the least range of dyed green. Conclusion:Didangtang for purging heat

dispersing blood stasis and dredging collaterals can reduce the myocardial inflammation of DM rats by inhibiting activation of NF-кB pathway.

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