健脾化瘀解毒方抑制PI3K/AKT/HIF-1α通路阻断胃癌前病变恶性进展的机制  

Mechanism of Jianpi Huayu Jiedu Formula on inhibition PI3K/AKT/HIF-1αpathway and blocking malignant progression of gastric precancerous lesion

作　　者：潘华峰[1] 袁冬生[1] 刘伟[1] 赵自明 郭廷洪 刘远亮 杨良俊 贺生才 刘洋[1] 何维 PAN Hua-feng;YUAN Dong-sheng;LIU Wei;ZHAO Zi-ming;GUO Ting-hong;LIU Yuan-liang;YANG Liang-jun;HE Sheng-cai;LIU Yang;HE Wei(Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Guangdong Second Traditional Chinese Medicine Hospital(Guangdong Province Engineering Technology Research Institute of TCM),Guangzhou 510095,China;University of Science and Technology of Macao,Macao 999078,China)

机构地区：[1]广州中医药大学,广州510006 [2]广东省第二中医院(广东省中医药工程技术研究院),广州510095 [3]澳门科技大学,澳门999078

出　　处：《中华中医药杂志》2020年第6期2786-2790,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81473620,No.81673946);广东省自然科学基金项目(No.2017A030313845)。

摘　　要：目的:探讨健脾化瘀解毒方对磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKT)/缺氧诱导因子-1α(HIF-1α)信号通路的影响,分析其治疗胃癌前病变(GPL)的机制。方法:以同窝野生型小鼠为空白组,将10周龄Atp4a^(-/-)小鼠随机均分为模型组,维酶素组,健脾化瘀解毒方高、低剂量组,连续灌胃给药10周后,观察胃黏膜病理改变,采用Western Blot检测PI3K、p-AKT、HIF-1α和乳酸脱氢酶A(LDHA)蛋白的表达,免疫组化检测MUC2和Ki-67蛋白的表达。结果:光镜下可见模型组小鼠胃黏膜萎缩、肠上皮化生、异型增生性改变,而健脾化瘀解毒方各剂量组小鼠胃黏膜萎缩、肠上皮化生、组织异型性病变程度与范围均较模型组明显改善。与空白组比较,模型组胃黏膜PI3K、p-AKT、HIF-1α和LDHA蛋白表达升高(P<0.01);与模型组比较,健脾化瘀解毒方各剂量组PI3K、p-AKT、HIF-1α和LDHA蛋白表达均降低(P<0.01,P<0.05)。免疫组化结果显示,与空白组比较,模型组胃黏膜MUC2和Ki-67平均光密度和累积面积百分率均升高(P<0.01);与模型组比较,健脾化瘀解毒方高剂量组MUC2和Ki-67的平均光密度和累积面积百分率均降低(P<0.01),健脾化瘀解毒方低剂量组的MUC2累积面积百分率和Ki-67的平均光密度降低(P<0.01)。结论:健脾化瘀解毒方可抑制PI3K/AKT/HIF-1α信号通路的激活,抑制有氧糖酵解代谢产物LDHA表达,阻断细胞的恶性增殖与肠上皮化生,改善局部缺氧微环境,延缓GPL恶变。Objective:To explore the effects of Jianpi Huayu Jiedu Formula(JPHY)on PI3K/AKT/HIF-1αsignal pathway and analyze its mechanism for treating gastric precancerous lesions(GPL).Methods:Using the same littermate of wildtype mice as the control group.Atp4a-/-mice at 10 weeks of age were randomly divided into the model group,the enzyme group,the JPHY high-dose(JPHY-H)and the JPHY low-dose(JPHY-L)group.The gastric mucosa was observed for 10 weeks after continuous intragastric administration.The expressions of PI3K,p-AKT,HIF-1α,and LDHA proteins were detected by Western Blot,and the MUC2 and Ki-67 protein expression were detected by immunohistochemistry.Results:Under the light microscope,the gastric mucosa atrophy,intestinal,and dysplasia of model group mice were observed,while the extent of atrophy,intestinal metaplasia,and dysplasia of gastric mucosa in the JPHY group were significantly improved compared with that in the model group.Compared with the control group,PI3K,p-AKT,HIF-1αand LDHA protein expression of gastric mucosa were increased in the model group(P<0.01).Compared with the model group,the PI3K,p-AKT,HIF-1αand LDHA protein in the JPHY groups were decreased(P<0.01,P<0.05).The immunohistochemical results showed that compared with the control group,the average optical density and cumulative area percentage of gastric mucosa MUC2 and Ki-67 in the model group were increased(P<0.01);Compared with the model group,the average optical density and cumulative area percentage of MUC2 and Ki-67 in the JPHY-H group were decreased(P<0.01),and the cumulative area percentage of MUC2 and average optical density of Ki-67 in the JPHY-L group were decreased(P<0.01).Conclusion:Jianpi Huayu Jiedu Formula can inhibit the activation of PI3K/AKT/HIF-1αsignaling pathway,inhibit the expression of aerobic glycolysis metabolite LDHA,block the malignant proliferation and intestinalization of cells,protect the gastric mucosa,and improve local hypoxia microenvironment,delay the malignant change of GPL.

关 键 词：胃癌前病变 健脾化瘀解毒方 磷脂酰肌醇-3-激酶 蛋白激酶B 缺氧诱导因子-1α 缺氧微环境 有氧糖酵解 Atp4a基因敲除小鼠 

分 类 号：R285.5[医药卫生—中药学]