三仁汤对幽门螺杆菌相关性胃炎脾胃湿热证大鼠消化吸收功能及Akt/NF-κB通路的影响

Effect of San Ren decoction on digestion and absorption function and Akt/NF-κB pathway in rats with H. pylori-associated gastritis and spleen and stomach damp heat syndrome

关键词:  幽门螺杆菌相关性胃炎  脾胃湿热证  三仁汤  Akt/NF-κB通路

Key words:  helicobacter pylori-associated gastritis  spleen and stomach damp heat syndrome  San Ren decoction  Akt/NF-κB pathway

摘要:

摘要：目的 探讨三仁汤对幽门螺杆菌相关性胃炎脾胃湿热证大鼠消化吸收功能及Akt/NF-κB通路的影响。方法 72只Wistar大鼠随机分为空白组（n=12）和造模组（n=60），以复合因素（劳倦+饮食+苦寒药+环境+生物因子（Hp菌液））构建幽门螺杆菌相关性胃炎（HAG）脾胃湿热证大鼠模型。造模成功后，将造模大鼠随机分为模型组、四联疗法组（奥美拉唑2 mg/kg+阿莫西林100 mg/kg+克拉霉素50 mg/kg+胶体果胶秘胶囊35 mg/kg）、三仁汤低剂量组（3 g/kg）、三仁汤中剂量组（7.5 g/kg）、三仁汤高剂量组（15 g/kg），各组连续给药21d。实验过程中观察大鼠一般情况；快速尿素酶实验检测Hp定植；HE染色观察胃黏膜炎症情况；TUNEL检测胃黏膜组织细胞凋亡；ELISA检测血清Ghrelin、IFN-γ、MTL、GAS、IL-4、IL-10含量；Western blot检测胃黏膜组织caspase3、Bax、Bcl-2、cAMP、Akt、p-Akt、NF-κB p65、p-NF-κB p65蛋白表达；Real-time PCR检测胃黏膜组织Akt、NF-κB p65 mRNA表达。结果 与空白组比较，模型组大鼠黏膜组织明显损伤，Hp定植率、细胞凋亡、胃黏膜组织caspase3、Bax、p-Akt、p-NF-κB p65蛋白表达、胃黏膜组织Akt、NF-κB p65 mRNA表达及血清IFN-γ含量明显升高，胃黏膜组织Bcl-2蛋白表达及血清Ghrelin、MTL、GAS、IL-4、IL-10含量明显降低（P<0.01）；与模型组比较，三仁汤高剂量组大鼠黏膜组织损伤明显改善，Hp定植率、细胞凋亡、胃黏膜组织caspase3、Bax、p-Akt、p-NF-κB p65蛋白表达、胃黏膜组织Akt、NF-κB p65 mRNA表达及血清IFN-γ含量明显降低，胃黏膜组织Bcl-2蛋白表达及血清Ghrelin、MTL、GAS、IL-4、IL-10含量明显升高（P<0.05）。结论 三仁汤可改善HAG脾胃湿热证大鼠症状，抑制炎症反应及功能性消化不良，作用机制可能与调控Akt/NF-κB通路有关。

Abstract:

Objective To investigate the effects of San Ren decoction on digestion and absorption function and Akt/NF-κB pathway in rats with H. pylori-associated gastritis and spleen and stomach damp heat syndrome. Methods Seventy-two Wistar rats were randomly divided into control group (n=12) and modeling group (n=60), and a rat model of H. pylori-associated gastritis (HAG) with a combination of factors (fatigue + diet+ bitter cold medicine +environment +biological factors (Hp bacterial solution)) was constructed. After successful modeling, the rats were randomly divided into model group, quadruple therapy group (omeprazole 2 mg/kg + amoxicillin 100 mg/kg + clarithromycin 50 mg/kg + colloidal pectin secret capsule 35 mg/kg), San Ren decoction low dose group (3 g/kg), San Ren decoction medium dose group (7.5 g/kg), San Ren decoction high dose group (15 g/kg), and each group was continuously administered for 21 d. During the experiment, the general conditions of rat were observed; the rapid urease test was performed to detect Hp colonization; HE staining was performed to observe the inflammation of gastric mucosa; TUNEL was performed to detect apoptosis in gastric mucosa; the content of Ghrelin, IFN-γ, MTL, GAS, IL-4, IL-10 in serum were detected by ELISA.; Western blot was used to detect the protein expression of caspase3, Bax, Bcl-2, cAMP, Akt, p-Akt, NF-κB p65, p-NF-κB p65 in gastric mucosa. Real-time PCR was performed to detect Akt, NF-κB p65 mRNA expression in gastric mucosal tissues. Results Compared with the control group, the mucosal tissue of rats in the model group was significantly damaged, and the Hp colonization rate, apoptosis, protein expression of caspase3, Bax, p-Akt, p-NF-κB p65 in gastric mucosal tissue, mRNA expression of Akt, NF-κB p65 gastric mucosal tissue and serum IFN-γ content were significantly increased, gastric mucosal tissue Bcl-2 protein expression and serum Ghrelin, MTL, GAS, IL-4 and IL-10 levels were significantly decreased in gastric mucosal tissues in the model group (P<0.01). Compared with the model group, the mucosal tissue injury of rats in San Ren decoction high dose group was significantly improved, the Hp colonization rate, apoptosis, protein expression of caspase3, Bax, p-Akt, p-NF-κB p65 in gastric mucosal tissues, mRNA expression of Akt and NF-κB p65 in gastric mucosal tissues and serum IFN-γ content were significantly reduced, and the protein expression of Bcl-2 in gastric mucosal tissues and serum levels of Ghrelin, MTL, GAS, IL-4 and IL-10 were significantly increased in San Ren decoction high dose group (P<0.05). Conclusion San Ren decoction can improve the symptoms, inhibit the inflammatory response and functional dyspepsia in rats with HAG spleen and stomach damp heat syndrome, and the mechanism of action may be related to the regulation of Akt/NF-κB pathway.