基于MCT4/CD147探讨四君子汤加减改善酸性微环境逆转胃癌前病变的效应机制

目的:观察四君子汤加减治疗胃癌前病变(GPL)模型大鼠胃黏膜乳酸水平及单羧酸转运蛋白1(MCT1),单羧酸转运蛋白4(MCT4)和细胞外基质金属蛋白酶诱导物(CD147)表达的影响。方法:将74只SD雄性大鼠随机分为正常组12只,造模组62只。采用N-甲基-N’-硝基-N-亚硝基胍(MNNG)-氨水复合造模法诱导制作GPL大鼠模型,于第9周将造模组大鼠随机分为模型组,叶酸组(2.7 mg·kg^-1),四君子汤加减高、中、低剂量组(12.6,6.3,3.15 g·kg^-1),每组12只,灌胃治疗持续12周,观察大鼠一般情况。采用苏木素-伊红(HE)染色观察大鼠胃黏膜组织病理学改变;化学比色法检测胃黏膜组织乳酸含量;免疫组化和实时荧光定量聚合酶链式反应(Real-time PCR)检测胃黏膜组织MCT1,MCT4,CD147蛋白和mRNA表达。结果:四君子汤加减可显著改善GPL大鼠胃黏膜上皮腺体结构、排列紊乱和细胞异型性等病理表现。与正常组比较,模型组大鼠胃黏膜组织乳酸含量显著升高(P<0.01),MCT1,MCT4,CD147蛋白与mRNA表达均明显升高(P<0.05,P<0.01);与模型组比较,四君子汤加减各剂量组乳酸含量均明显降低(P<0.05,P<0.01),四君子汤加减高、中、低剂量组MCT4和CD147蛋白表达均显著降低(P<0.05,P<0.01),四君子汤加减高、中剂量组MCT4 mRNA表达明显降低(P<0.05,P<0.01),四君子汤加减高剂量组CD147mRNA表达明显降低(P<0.05)。四君子汤加减对MCT1无显著调控作用。结论:四君子汤加减可显著改善GPL模型大鼠胃黏膜上皮异常组织病理学表现,其机制可能与下调MCT4和CD147过度表达,抑制乳酸外流,改善胃黏膜上皮酸性微环境有关。

Explore Mechanism of Modified Si Junzitang in Improving Acidic Microenvironment and Reversing Gastric Precancerous Lesions Based on MCT4/CD147

Objective:To observe the effect of modified Si Junzitang on the level of lactic acid in gastric mucosa and the expression of Carboxylic acid transporter 1(MCT1),monocarboxylic acid transporter 4(MCT4),and extracellular matrix metalloproteinase inducer(CD147)in rats with gastric precancerous lesions(GPL).Method:Seventy-four SD male rats were randomly divided into normal group(12 rats)and model group(62 rats).N-methyl-N’-nitro-N-nitrosoguanidine(MNNG)-ammonia compound method was used to establish GPL rat models,and at the 9^thweek,the model rats were randomly divided into model group,folic acid group(2.7 mg·kg^-1),modified Si Junzitang high,medium and low dose groups(12.6,6.3,3.15 g·kg^-1),with 12 rats in each group.After intragastric administration for 12 weeks,the general conditions of the rats were observed.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of gastric mucosa in rats,chemical colorimetry was used to detect the content of lactic acid in gastric mucosa;immunohistochemistry and real-time polymerase chain reaction(Real-time PCR)were used to detect MCT1,MCT4,CD147 protein and mRNA expression in gastric mucosal tissues.Result:Modified Si Junzitang significantly improved the pathological manifestations in GPL rats such as gastric mucosal epithelial gland structure,disorder of arrangement and cell atypia.Compared with the normal group,the lactic acid content of the gastric mucosa tissue in the model group increased significantly(P<0.01),and the protein and mRNA expressions of MCT1,MCT4,CD147 significantly increased(P<0.05,P<0.01).Compared with the model group,the lactic acid content in each dose group of modified Si Junzitang was significantly reduced(P<0.05,P<0.01),and the protein expression levels of MCT4 and CD147 were also significantly reduced in each dose group of modified Si Junzitang(P<0.05,P<0.01).The mRNA expression of MCT4 was significantly reduced in the middle and high dose groups(P<0.05,P<0.01),and the mRNA expression of CD147 was significantly reduced in the high dose group(P<0.05).Modified Si Junzitang showed no significant regulatory effect on MCT1.Conclusion:Modified Si Junzitang can significantly improve the abnormal histopathology of gastric mucosal epithelium in GPL model rats.Its mechanism may be related to down-regulating the overexpression of MCT4 and CD147,inhibiting lactic acid outflow,and improving the acidic microenvironment of gastric mucosal epithelium.