香砂六君子汤对幽门螺杆菌诱导的慢性萎缩性胃炎大鼠胃黏膜组织TLR信号通路的影响

[Effects of Xiangsha Liujunzi decoction on TLR signal pathway in gastric mucosa tissues of rats with Helicobacter pylori-induced chronic atrophic gastritis].

作者信息

Lin Zhi-Qiang, Wang Da-Xuan, Hong Shan-Shan, Fu Xin-Yang

机构信息

Department of Pharmacy, the First Hospital of Quanzhou Affiliated to Fujian Medical University, Quanzhou 362000, China.

Department of Respiration, the First Hospital of Quanzhou Affiliated to Fujian Medical University, Quanzhou 362000, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2016 Aug;41(16):3078-3083. doi: 10.4268/cjcmm20161623.

DOI:10.4268/cjcmm20161623

PMID:28920352

Abstract

To explore the effects and mechanism of Xiangsha Liujunzi decoction on TLR signal pathway in gastric mucosa tissues of rats with Helicobacter pylori-related gastritis, sixty SD rats were randomly divided into control group, model group, high concentration of Xiangsha Liujunzi decoction group, moderate concentration of Xiangsha Liujunzi decoction group, low concentrations of Xiangsha Liujunzi decoction group and SB203580-treated group, with 10 rats in each group. SD rats of Hp-associated chronic atrophic gastritis models were established by intragastric gavage of Helicobacter pylori (HP) suspension. Changes in the gastric mucosa of rats were assessed by histopathology. ELISA was applied to detect the expressions of TNF-α and IL-6 in the serum, and the activity of iNOS in gastric mucosa. The content of NO in the gastric mucosa was tested by nitrate reductive enzymatic. The expressions of TLR2, TLR4, P38MAPK, NF-κB were detected by QPCR and Western-blot. The results indicated that the clinical symptoms of rats and pathological changes of gastric mucosa were improved in Xiangsha Liujunzi decoction group. Compared with normal control group, the protein expressions of TLR2, TLR4, p-P38MAPK and NF-κB in gastric mucosa of model group rats increased (P<0.01) with the levels of TNF-α and IL-6 in the serum, and the activity of iNOS and the content of NO in gastric mucosa increased. Compared with model group, the expressions decreased in Xiangsha Liujunzi decoction group, especially in the high concentration of Xiangsha Liujunzi decoction group(P<0.01), with gradually increased rate of HP eradication and decreased pathological grades of chronic atrophic gastritis. The serum level of TNF-α and IL-6 decreased from (24.313±2.261) μg•L ⁻¹ to (15.195±1.235) μg•L-1(P<0.01) and from (77.416±8.095) μg•L ⁻¹ to (33.150±2.532) μg•L ⁻¹ (P<0.01), and the activity of iNOS and the content of NO in gastric mucosa decreased from (1.530±0.206) U•mg ⁻¹ to (0.802±0.091) U•mg ⁻¹ (P<0.01) and from (0.907±0.032) mmol•g ⁻¹ to (0.335±0.026) mmol•g ⁻¹ (P<0.01) after the treatment of high concentration of Xiangsha Liujunzi decoction. All the effects increased with the increasing dosage of Xiangsha Liujunzi decoction from 0.324 g•mg ⁻¹ to 1.296 g•mg ⁻¹. The protein expressions of NF-κB decreased in the gastric mucosa after treated with P38MAPK specific inhibitor-SB203580. In the rats model, HP infection results in chronic atrophic gastritis through the activation of TLR2, TLR4/MAPK/NF-κB/iNOS/NO signal pathway. Xiangsha Liujunzi decoction can eradicate H. pylori and alleviate chronic atrophic gastric mucosal inflammation. The treatment is effective and safe to cure HP-induced chronic atrophic gastritis.

摘要

为探讨香砂六君子汤对幽门螺杆菌相关性胃炎大鼠胃黏膜组织TLR信号通路的影响及作用机制，将60只SD大鼠随机分为对照组、模型组、香砂六君子汤高浓度组、香砂六君子汤中浓度组、香砂六君子汤低浓度组和SB203580处理组，每组10只。通过灌喂幽门螺杆菌（HP）悬液建立HP相关性慢性萎缩性胃炎SD大鼠模型。采用组织病理学评估大鼠胃黏膜的变化。应用ELISA法检测血清中TNF-α和IL-6的表达以及胃黏膜中iNOS的活性。采用硝酸还原酶法检测胃黏膜中NO的含量。通过QPCR和Western-blot检测TLR2、TLR4、P38MAPK、NF-κB的表达。结果表明，香砂六君子汤组大鼠的临床症状及胃黏膜病理改变均得到改善。与正常对照组相比，模型组大鼠胃黏膜中TLR2、TLR4、p-P38MAPK和NF-κB的蛋白表达升高（P<0.01），血清中TNF-α和IL-6水平、胃黏膜中iNOS活性及NO含量升高。与模型组相比，香砂六君子汤组上述表达降低，尤以香砂六君子汤高浓度组明显（P<0.01），HP根除率逐渐升高，慢性萎缩性胃炎病理分级降低。高浓度香砂六君子汤治疗后，血清TNF-α水平从（24.313±2.261）μg•L⁻¹降至（15.195±1.235）μg•L⁻¹（P<0.01），IL-6水平从（77.416±8.095）μg•L⁻¹降至（33.150±2.532）μg•L⁻¹（P<0.01），胃黏膜中iNOS活性从（1.530±0.206）U•mg⁻¹降至（0.802±0.091）U•mg⁻¹（P<0.01），NO含量从（0.907±0.032）mmol•g⁻¹降至（0.335±0.026）mmol•g⁻¹（P<0.01）。随着香砂六君子汤剂量从0.（此处原文有误，推测为0.324）g•mg⁻¹增加至1.296 g•mg⁻¹，上述各项作用增强。用P38MAPK特异性抑制剂SB203580处理后，胃黏膜中NF-κB蛋白表达降低。在大鼠模型中，HP感染通过激活TLR2、TLR4/MAPK/NF-κB/iNOS/NO信号通路导致慢性萎缩性胃炎。香砂六君子汤可根除HP并减轻慢性萎缩性胃黏膜炎症。该疗法治疗HP诱导的慢性萎缩性胃炎有效且安全。