Effect of Huanglian Wendantang on Inflammatory Response and Hepatocyte Burn Death in IGT Rats

Han LI; Yun-lei HUANG; Wan-ru DONG; Bo-yan MA

doi:10.13422/j.cnki.syfjx.20210507

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Effect of Huanglian Wendantang on Inflammatory Response and Hepatocyte Burn Death in IGT Rats

Classic Prescriptions | 更新时间：2021-03-29

- Effect of Huanglian Wendantang on Inflammatory Response and Hepatocyte Burn Death in IGT Rats
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 27, Issue 8, Pages: 25-32(2021)
- 作者机构：
  
  1.什邡市人民医院，四川什邡 618400
  2.空军航空大学，长春 130000
  3.黑龙江中医药大学药物安全性评价中心，哈尔滨 150040
  4.黑龙江中医药大学，哈尔滨 150040
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20210507
  CLC： R2-0;R22;R285.5;R289
- Received：15 December 2020，
  
  Published Online：26 February 2021，
  
  Published：20 April 2021
- 稿件说明：
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李寒,黄云蕾,董婉茹等.黄连温胆汤对IGT大鼠炎症反应与肝细胞焦亡的影响[J].中国实验方剂学杂志,2021,27(08):25-32.

LI Han,HUANG Yun-lei,DONG Wan-ru,et al.Effect of Huanglian Wendantang on Inflammatory Response and Hepatocyte Burn Death in IGT Rats[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(08):25-32.
李寒,黄云蕾,董婉茹等.黄连温胆汤对IGT大鼠炎症反应与肝细胞焦亡的影响[J].中国实验方剂学杂志,2021,27(08):25-32. DOI： 10.13422/j.cnki.syfjx.20210507.

LI Han,HUANG Yun-lei,DONG Wan-ru,et al.Effect of Huanglian Wendantang on Inflammatory Response and Hepatocyte Burn Death in IGT Rats[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(08):25-32. DOI： 10.13422/j.cnki.syfjx.20210507.

摘要

目的

探讨高脂饮食诱导糖耐量低减（IGT）大鼠是否存在炎症反应与细胞焦亡及黄连温胆汤对其干预作用。

方法

健康雄性SD大鼠予45%脂肪含量饲料喂养20周复制IGT模型。将符合模型标准的大鼠随机分为3组，每组10只，另取10只空白鼠作为正常组。黄连温胆汤组予7.8 g·kg

-1

·d

-1

复方水煎液，阳性药组予盐酸二甲双胍水溶液0.05 g·kg

-1

·d

-1

，体积为10 mL·kg

-1

·d

-1

，正常组和模型组予以同体积蒸馏水。连续灌胃4周后，测体质量、体长、腹围，计算Lee's指数；测空腹血浆葡萄糖（FPG）及餐后2 h血糖（2 h PG）。酶联免疫吸附测定（ELISA）检测血清白介素-6（IL-6）含量；实时荧光定量聚合酶链式反应（Real-time PCR）和蛋白免疫印迹法（Western blot）测肝组织核转录因子-

B（NF-

B），天冬氨酸特异性半胱氨酸蛋白酶-1（Caspase-1）及NOD样受体蛋白3（NLRP3） mRNA及蛋白表达；免疫荧光法测肝组织Caspase-1及NLRP3蛋白表达；苏木素-伊红（HE）染色观察肝组织形态。

结果

与正常组比较，模型组大鼠体质量、腹围、体长及Lee's指数明显增长（

0.05，

0.01），FPG及2 h PG水平均明显升高（

0.05，

0.01），血清中IL-6含量和肝组织NF-

B，Caspase-1及NLRP3基因与蛋白表达均显著增加（

0.01），免疫荧光显示IGT大鼠肝组织Caspase-1和NLRP3表达明显上调，HE染色见IGT大鼠肝组织存在炎症细胞浸润；与模型组比较，黄连温胆汤和盐酸二甲双胍组均可有效减轻IGT大鼠体质量（

0.01），腹型肥胖，纠正FPG和2 h PG水平（

0.01），同时能有效降低IGT大鼠血清IL-6含量和肝组织中NF-

B，Caspase-1及NLRP3基因与蛋白表达，减轻肝组织炎症细胞浸润程度。

结论

高脂饮食诱导IGT大鼠存在明显炎症反应及细胞焦亡，黄连温胆汤可明显减轻其炎症反应，缓解肝细胞焦亡。

Abstract

Objective

To investigate whether there is inflammatory reaction and cell pyroptosis in impaired glucose tolerance （IGT） rats induced by high-fat diet and the intervention effect of Huanglian Wendantang.

Method

Healthy male SD rats were fed with 45% fat content diet for 20 weeks to replicate the IGT model. The rats in line with the model establish criteria were randomly divided into 3 groups， with 10 rats in each group， another 10 rats were selected as the blank control group. Huanglian Wendantang group was given 7.8 g·kg

-1

·d

-1

compound decoction of Huanglian Wendantang， and the positive control group was given 0.05 g·kg

-1

·d

-1

aqueous solution of metformin hydrochloride， with the dose volume of 10 mL·kg

-1

·d

-1

. The blank group and the model group were given the same volume of distilled water. After continuous intragastric administration for 4 weeks， the body weight， body length and abdominal circumferences were measured， the Lee's obesity index was calculated， and the levels of fasting plasma glucose （FPG） and 2-hour plasma glucose （2 h PG） were detected in each group. Serum interleukin-6 （IL-6） content was detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expressions of nuclear factor kappa B （NF-

B）， cysteinyl aspartate specific proteinase-1 （Caspase-1） and NOD-like receptor protein 3 （NLRP3） in liver tissues were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. Immunofluorescence was used to detect the expression of Caspase-1 and NLRP3 in rat liver. Hematoxylin and eosin （HE） staining was used to observe the morphology of liver tissue.

Result

Compared with blank group， the body weight， abdominal circumference， body length and Lee's index in model group were significantly increased （

0.05，

0.01）， the levels of FPG and 2 h PG were significantly increased （

0.05，

0.01）， serum IL-6 content and NF-

B， Caspase-1 and NLRP3 gene and protein expressions in liver tissue were significantly increased （

0.01）， immunofluorescence technique showed that Caspase-1 and NLRP3 expressions increased obviously in IGT rat model’s liver tissues， and inflammatory cells infiltration was observed obviously in HE stained rat liver tissue model. Compared with model group， Huanglian Wendantang and metformin hydrochloride groups could effectively reduce the body weight of IGT rats （

0.01） and abdominal obesity， and correct the levels of FPG and 2 h PG （

0.01）， while effectively reducing serum IL-6 content and gene and protein expressions of NF-

B， Caspase-1 and NLRP3 in liver tissues of IGT rats， and alleviating inflammatory cell infiltration.

Conclusion

Inflammatory response exists in IGT model rats induced by high-fat diet. Huanglian Wendantang can obviously reduce its inflammatory response and alleviate liver cell pyroptosis.

关键词

Keywords

references

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