Yin Jing, Yi Jinyu, Yang Chun, Xu Bo, Lin Jiang, Hu Hongyi, Wu Xiaojun, Shi Hailian, Fei Xiaoyan
Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China.
Shanghai Key Laboratory of Compound Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
Evid Based Complement Alternat Med. 2019 Jun 20;2019:2651037. doi: 10.1155/2019/2651037. eCollection 2019.
Chronic atrophic gastritis (CAG), the precancerous lesions of gastric cancer, plays an important role in the stepwise process of gastric cancer. The ancient Chinese medicine believes in that Qi deficiency and blood stasis are involved in the pathogenesis of CAG. Weiqi decoction, a classical formula from Longhua Hospital, could supplement Qi and activate blood circulation of human beings and has been used for treating CAG in clinic over twenty years. The study aims to clarify the effect and underlying molecular mechanism of Weiqi decoction on CAG rats.
Forty-eight male Wistar rats were divided randomly into six groups: control group, model group, folic acid group, and WQD-treated groups at doses of 4 g/kg, 2 g/kg, and 1 g/kg, with eight rats in each group. MNNG and saturated NaCl were used to induce CAG rat with precancerous lesion (intestinal metaplasia and dysplasia). After 40 weeks, gastric mucosal blood flow was measured using Laser Doppler Flowmetry. The pathological changes of the gastric mucosa were identified by H&E staining and AB-PAS staining. The protein expression of COX-2, HIF-1, VEGFR1, VEGFR2, Ki67, and cleaved caspase 3 in the gastric tissues was measured by western blotting approach. Gene expression of COX-2, HIF-1, VEGF, VEGFR1, VEGFR2, Ang-1, and Ang-2 was detected by using Quantitative PCR method. The PGE2 concentrations in serum were detected by ELISA method. The protein expression of Ki67 in gastric mucosa was also detected by immunohistochemistry.
Compared with control rats, atrophy and intestinal metaplasia as well as the microcirculation disturbance of gastric mucosa were induced in the stomach of CAG rats identified by the H&E and AB-PAS staining as well as microcirculation measurement, which could be significantly attenuated by WQD treatment. Moreover, compared with the control group, the protein and gene expression of COX-2, HIF-1, VEGFR1, and VEGFR2 in gastric tissues of pylorus was obviously increased and the serum PGE2 level was significantly deceased in CAG rats, which could be significantly counteracted by WQD administration. However, the gene expression of Ang-1 and Ang-2 was not significant difference between control rats and CAG rats, and WQD also had no significant effect on the gene expression of Ang-1 and Ang-2. Furthermore, the increased cell proliferation marked by upregulated protein expression of Ki67 and decreased cell apoptosis marked by downregulated protein expression of cleaved caspase 3 in stomach of pylorus in CAG rats were obviously reversed by WQD treatment.
WQD attenuated CAG with precancerous lesion through regulating gastric mucosal blood flow disturbance and HIF-1 signaling pathway.
慢性萎缩性胃炎（CAG）作为胃癌的癌前病变，在胃癌的逐步发展过程中起重要作用。中医认为气虚血瘀参与了CAG的发病机制。萎芪汤是龙华医院的经典方剂，能补气活血，已在临床上用于治疗CAG二十余年。本研究旨在阐明萎芪汤对CAG大鼠的作用及其潜在分子机制。
48只雄性Wistar大鼠随机分为6组：对照组、模型组、叶酸组以及萎芪汤高剂量（4 g/kg）、中剂量（2 g/kg）、低剂量（1 g/kg）组，每组8只。采用MNNG和饱和NaCl诱导大鼠发生伴有癌前病变（肠化生和发育异常）的CAG。40周后，用激光多普勒血流仪测量胃黏膜血流量。通过苏木精-伊红（H&E）染色和AB-PAS染色鉴定胃黏膜的病理变化。采用蛋白质印迹法检测胃组织中COX-2、HIF-1、VEGFR1、VEGFR2、Ki67和裂解型半胱天冬酶3的蛋白表达。采用定量PCR法检测COX-2、HIF-1、VEGF、VEGFR1、VEGFR2、Ang-1和Ang-2的基因表达。采用酶联免疫吸附测定（ELISA）法检测血清中PGE2浓度。采用免疫组织化学法检测胃黏膜中Ki67的蛋白表达。
与对照大鼠相比，H&E染色、AB-PAS染色及微循环检测显示CAG大鼠胃黏膜出现萎缩、肠化生及微循环障碍，萎芪汤治疗可明显减轻上述病变。此外，与对照组相比，CAG大鼠幽门部胃组织中COX-2、HIF-1、VEGFR1和VEGFR2的蛋白及基因表达明显升高，血清PGE2水平明显降低，萎芪汤给药可显著对抗上述变化。然而，对照大鼠与CAG大鼠之间Ang-1和Ang-2的基因表达无显著差异，萎芪汤对Ang-1和Ang-2的基因表达也无显著影响。此外，萎芪汤治疗明显逆转了CAG大鼠幽门部胃组织中以Ki67蛋白表达上调为标志的细胞增殖增加及以裂解型半胱天冬酶3蛋白表达下调为标志的细胞凋亡减少。
萎芪汤通过调节胃黏膜血流紊乱和HIF-1信号通路减轻伴有癌前病变的CAG。
