Study on the mechanism of Yinxu Weitong capsule in the treatment of chronic atrophic gastritis rats based on HIF-1α/GLUT-1 pathway
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摘要: 目的 探讨阴虚胃痛胶囊通过调控缺氧诱导因子-1α(hypoxia inducible factor-1α，HIF-1α)/葡萄糖转运蛋白-1(glucose transporter-1，GLUT-1)通路治疗慢性萎缩性胃炎(chronic atrophic gastritis，CAG)大鼠的作用机制。方法 采用N-甲基-N’-硝基-N-亚硝基胍(MNNG)自由饮+灌胃，联合饥饱失常法复合因素干预制备CAG模型。成功建模后，随机将建模大鼠分为模型组及低、中、高剂量阴虚胃痛胶囊组和维酶素组，每组均10只。阴虚胃痛胶囊组按照不同剂量分别给予不同浓度的药液灌胃，维酶素组给予维酶素溶液灌胃，正常组和模型组则给予生理盐水灌胃，灌胃体积均为10 mL/kg，给药8周。利用苏木精-伊红染色法观察大鼠胃黏膜病理变化，使用qRT-PCR和Western blot法检测大鼠胃黏膜中HIF-1α、GLUT-1的mRNA和蛋白表达水平，采用Western blot法检测HK2、PFKM、PKM2、LDHA蛋白表达水平。结果 与正常组比较，模型组大鼠胃黏膜腺体明显减少，HIF-1α、GLUT-1mRNA及蛋白表达水平显著升高(P < 0.01)，HK2、PFKM、PKM2、LDHA蛋白表达水平显著升高(P < 0.05)。经低、中、高剂量阴虚胃痛胶囊治疗后，大鼠胃黏膜腺体较前恢复，HIF-1α、GLUT-1mRNA及HIF-1α、GLUT-1、HK2、PKM2蛋白表达水平明显降低(P < 0.05，P < 0.01，P < 0.001)；经中、高剂量阴虚胃痛胶囊治疗后，PFKM、LDHA蛋白表达水平明显降低(P < 0.01)。结论 阴虚胃痛胶囊可有效改善CAG大鼠胃黏膜病变，其作用机制可能与调控HIF-1α/GLUT-1通路相关。Abstract: Objective To investigate the mechanism of Yinxu Weitong capsule on chronic atrophic gastritis(CAG) rats by regulating hypoxia inducible factor-1α(HIF-1α)/glucose transporter-1(GLUT-1) pathway.Methods CAG model was established by free drinking and gavage of N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) combined with compound factors intervention of disturbance of appetite and satiety. After successful modeling, rats in the modeling group were randomly divided into model group, Yinxu Weitong capsule low-dose, medium-dose, high-dose groups and vitamin E group, with 10 rats in each group. Yinxu Weitong capsule low-dose, medium-dose, high-dose groups were given corresponding doses of liquid gavage; positive drug group was given vitamin E suspension gavage, normal group and model group were given saline gavage. The gavage volume was 10 mL/kg, and the drug was given for 8 weeks. The pathological changes of gastric mucosa in each group of CAG rats were observed by HE staining. The mRNA and protein expression levels of HIF-1α, GLUT-1 in gastric mucosa of each group of CAG rats were detected by qRT-PCR and Western blot methods, and the protein expression levels of HK2, PFKM, PKM2 and LDHA were detected by Western blot method.Results Compared with the normal group, the gastric mucosal glands of rats in the model group were significantly reduced, the mRNA content and protein expression of HIF-1α and GLUT-1 were significantly increased(P < 0.01), and the protein expression of HK2, PFKM, PKM2, LDHA were significantly increased(P < 0.05). After treatment with low, medium and high dose Yinxu Weitong capsule, the gastric mucosal glands were recovered, the mRNA contents of HIF-1α and GLUT-1 and the protein expressions of HIF-1α, GLUT-1, HK2 and PKM2(P < 0.05, P < 0.01, P < 0.001) were significantly decreased, and the protein levels of PFKM and LDHA were significantly decreased after treatment with medium and high dose Yinxu Weitong capsule(P < 0.01).Conclusion Yinxu Weitong capsule can effectively improve gastric mucosal lesions in CAG rats, and its mechanism may be related to the regulation of HIF-1α/GLUT-1 pathway.
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表 1 本实验使用的引物序列
引物名称 引物序列(上游) 引物序列(下游) HIF-1α ACCGTGCCCCTACTATGTCG GCCTTGTATGGGAGCATTAACTT GLUT-1 AGGAGATGAAAGAAGAGGGTCG GTGTTGACGATACCCGAGCC GAPDH CTGGAGAAACCTGCCAAGTATG GGTGGAAGAATGGGAGTTGCT 
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表 2 本实验使用的抗体及稀释倍数
抗体名称 稀释倍数 生产厂家 货号 兔HIF-1α抗体 1：1 000 Immunoway YT2133 兔GLUT-1抗体 1：100 000 Abcam 115730 鼠PKM2抗体 1：5 000 Proteintech 60268-1-AP 兔LDHA抗体 1：5 000 Proteintech 19987-1-AP 兔HK2抗体 1：5 000 Proteintech 22029-1-AP 兔PFKM抗体 1：2 000 Proteintech 55028-1-AP 兔β-actin抗体 1：1 000 CST 4970S
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[1] Shah SC, Piazuelo MB, Kuipers EJ, et al. AGA Clinical Practice Update on the Diagnosis and Management of Atrophic Gastritis: Expert Review[J]. Gastroenterology, 2021, 161(4): 1325-1332. e7. doi: 10.1053/j.gastro.2021.06.078
[2] Lahner E, Zagari RM, Zullo A, et al. Chronic atrophic gastritis: Natural history, diagnosis and therapeutic management. A position paper by the Italian Society of Hospital Gastroenterologists and Digestive Endoscopists[AIGO], the Italian Society of Digestive Endoscopy[SIED], the Italian Society of Gastroenterology[SIGE], and the Italian Society of Internal Medicine[SIMI][J]. Dig Liver Dis, 2019, 51(12): 1621-1632. doi: 10.1016/j.dld.2019.09.016
[3] Correa P. Human gastric carcinogenesis: a multistep and multifactorial process-First American Cancer Society Award Lecture on Cancer Epidemiology and Prevention[J]. Cancer Res, 1992, 52(24): 6735-6740.
[4] Yang T, Wang R, Liu H, et al. Berberine regulates macrophage polarization through IL-4-STAT6 signaling pathway in Helicobacter pylori-induced chronic atrophic gastritis[J]. Life Sci, 2021, 266: 118903. doi: 10.1016/j.lfs.2020.118903
[5] 房静远, 杜奕奇, 刘文忠, 等. 中国慢性胃炎共识意见(2017年, 上海)[J]. 胃肠病学, 2017, 22(11): 670-687. doi: 10.3969/j.issn.1008-7125.2017.11.007
[6] Luo C, Sun Z, Li Z, et al. Notoginsenoside R1(NGR1) Attenuates Chronic Atrophic Gastritis in Rats[J]. Med Sci Monit, 2019, 25: 1177-1186. doi: 10.12659/MSM.911512
[7] Fernandez-coto DL, Gil J, Hernandez A, et al. Quantitative proteomics reveals proteins involved in the progression from non-cancerous lesions to gastric cancer[J]. J Proteomics, 2018, 186: 15-27. doi: 10.1016/j.jprot.2018.07.013
[8] 刘伟. 胃痞灵调控mTOR/HIF-1α/SIRT6信号通路介导胃癌前病变有氧糖酵解的分子机制研究[D]. 广州: 广州中医药大学, 2020.
[9] 钱素婷. HIF-1α通路介导MNNG诱发新生大鼠胃癌前病变的分子机制及益气活血方的干预作用[D]. 杭州: 浙江中医药大学, 2023.
[10] Luo Z, Tian M, Yang G, et al. Hypoxia signaling in human health and diseases: implications and prospects for therapeutics[J]. Signal Transduct Target Ther, 2022, 7(1): 218. doi: 10.1038/s41392-022-01080-1
[11] Yin J, Yi J, Yang C, et al. Weiqi Decoction Attenuated Chronic Atrophic Gastritis with Precancerous Lesion through Regulating Microcirculation Disturbance and HIF-1alpha Signaling Pathway[J]. Evid Based Complement Alternat Med, 2019, 2019: 2651037.
[12] 钱胜. 阴虚胃痛颗粒联合三联疗法治疗阴虚型慢性胃炎疗效观察[J]. 现代中西医结合杂志, 2015, 24(5): 478-480. https://www.cnki.com.cn/Article/CJFDTOTAL-XDJH201505009.htm
[13] Tong Y, Wang R, Liu X, et al. Zuojin Pill ameliorates chronic atrophic gastritis induced by MNNG through TGF-beta1/PI3K/Akt axis[J]. J Ethnopharmacol, 2021, 271: 113893. doi: 10.1016/j.jep.2021.113893
[14] 张卫茜, 王瑞昕, 赵鲁卿, 等. 大黄三味片通过SCF/c-Kit信号通路对便秘大鼠肠道动力的影响[J]. 中国中西医结合消化杂志, 2023, 31(11): 855-860. doi: 10.3969/j.issn.1671-038X.2023.11.07
[15] 张声生, 唐旭东, 黄穗平, 等. 慢性胃炎中医诊疗专家共识意见(2017)[J]. 中华中医药杂志, 2017, 32(7): 3060-3064. https://www.cnki.com.cn/Article/CJFDTOTAL-BXYY201707063.htm
[16] 宋雅婵, 谢晶日, 李贺薇. 基于"阳化气, 阴成形"理论的慢性萎缩性胃炎辨治[J]. 时珍国医国药, 2018, 29(8): 1955-1956. https://www.cnki.com.cn/Article/CJFDTOTAL-SZGY201808057.htm
[17] 姬炫, 赵琦, 赵秦. 基于"甘寒生津"理论探讨胃阴虚型慢性萎缩性胃炎治疗经验[J]. 贵州中医药大学学报, 2023, 45(4): 5-8. https://www.cnki.com.cn/Article/CJFDTOTAL-GYZX202304002.htm
[18] Liu W, Pan HF, Yang LJ, et al. Panax ginseng C.A. Meyer(Rg3) Ameliorates Gastric Precancerous Lesions in Atp4a(-/-)Mice via Inhibition of Glycolysis through PI3K/AKT/miRNA-21 Pathway[J]. Evid Based Complement Alternat Med, 2020, 2020: 2672648.
[19] 张成哲, 卓俊城, 蔡甜甜, 等. 胃炎1号方对胃癌前病变大鼠胃黏膜上皮细胞缺氧及缺氧耐受的影响[J]. 中成药, 2017, 39(5): 896-901. https://www.cnki.com.cn/Article/CJFDTOTAL-ZCYA201705003.htm
[20] Kierans SJ, Taylor CT. Regulation of glycolysis by the hypoxia-inducible factor(HIF): implications for cellular physiology[J]. J Physiol, 2021, 599(1): 23-37.
[21] Li M, Li G, Yang X, et al. HIF in Gastric Cancer: Regulation and Therapeutic Target[J]. Molecules, 2022, 27(15): 4893.
[22] Tao J, Zhang Y, Wang T. Potential Role of Glucose Transporter-1 Expression in Gastric Cancer: A Meta-Analysis and Systematic Review[J]. Iran J Public Health, 2020, 49(11): 2044-2053.
[23] Wang J, Huang Q, Hu X, et al. Disrupting Circadian Rhythm via the PER1-HK2 Axis Reverses Trastuzumab Resistance in Gastric Cancer[J]. Cancer Res, 2022, 82(8): 1503-1517.
[24] Zhou Y, Lin F, Wan T, et al. ZEB1 enhances Warburg effect to facilitate tumorigenesis and metastasis of HCC by transcriptionally activating PFKM[J]. Theranostics, 2021, 11(12): 5926-5938.
[25] Gao X, Wang H, Yang JJ, et al. Pyruvate kinase M2 regulates gene transcription by acting as a protein kinase[J]. Mol Cell, 2012, 45(5): 598-609.
[26] Wang XH, Jiang ZH, Yang HM, et al. Hypoxia-induced FOXO4/LDHA axis modulates gastric cancer cell glycolysis and progression[J]. Clin Transl Med, 2021, 11(1): e279.
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