RESUMEN

Gastric cancer is recognized as one of the most common cancer. In-depth research of gastric precancerous lesions (GPL) plays an important role in preventing the occurrence of gastric cancer. Meanwhile, traditional treatment provides a novel sight in the prevention of occurrence and development of gastric cancer. The current study was designed to assess the effects of therapy with Weipixiao (WPX) decoction on N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced GPL rats and the underlying molecular mechanisms. After 10-weeks treatment, all rats were sacrificed. Histopathological changes of gastric tissue were assessed via hematoxylin-eosin (HE) and High-iron diamine-Alcian blue-Periodic acid-Schiff (HID-AB-PAS) staining. To be fully evidenced, RT-qPCR, Western blot and immunohistochemistry were used to detect the expressions of LDHA, CD147, HIF-1α, MCT4, PI3K, AKT, mTOR and miRNA-34a, which were crucial factors for evaluating GPL in the aspect of glycolysis pathogenesis. According to the results of HE and HID-AB-PAS staining, it could be confirmed that MNNG-induced GPL rats were obviously reversed by WPX decoction. Additionally, the increased gene levels of LDHA, CD147, MCT4, PI3K, AKT, mTOR and HIF-1α in model group were down-regulated by WPX decoction, while miRNA-34a expression was decreased and up-regulated by WPX decoction. The significantly increased protein levels of LDHA, CD147, MCT4, PI3K, AKT, mTOR and HIF-1α induced by MNNG were attenuated in rats treated with WPX decoction. In brief, the findings of this study imply that abnormal glycolysis in MNNG-induced GPL rats was relieved by WPX decoction via regulation of the expressions of LDHA, CD147, HIF-1α, MCT4, PI3K, AKT, mTOR and miRNA-34a.

Asunto(s)

Anticarcinógenos/farmacología , Medicamentos Herbarios Chinos/farmacología , Glucólisis/efectos de los fármacos , Metilnitronitrosoguanidina , Lesiones Precancerosas/prevención & control , Neoplasias Gástricas/prevención & control , Estómago/efectos de los fármacos , Animales , Citoprotección , Modelos Animales de Enfermedad , Regulación Enzimológica de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Masculino , Lesiones Precancerosas/inducido químicamente , Lesiones Precancerosas/enzimología , Lesiones Precancerosas/patología , Ratas Sprague-Dawley , Transducción de Señal , Estómago/enzimología , Estómago/patología , Neoplasias Gástricas/inducido químicamente , Neoplasias Gástricas/enzimología , Neoplasias Gástricas/patología

RESUMEN

BACKGROUND: Angiogenesis is a pathobiological hallmark of gastric cancer. However, rare studies focus on angiogenesis in gastric precancerous lesions (GPL). Weipixiao (WPX), a Chinese herbal preparation, is proved clinically effective in treating GPL. Here, we evaluated WPX's anti-angiogenic potential for GPL, and also investigated the possibility of its anti-angiogenic mechanisms. METHODS: HPLC analysis was applied to screen the major chemical components of WPX. After modeling N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced GPL in male Sprague-Dawley rats, different doses of WPX were administrated orally for 10 weeks. Next, we performed histopathological examination using routine H&E staining and HID-AB-PAS staining. In parallel, we assessed angiogenesis revealed by microvessel density (MVD) using CD34 immunostaining, and subsequently observe microvessel ultrastructure in gastric mucosa under Transmission Electron Microscope. Finally, we detect expression of angiogenesis-associated markers VEGF and HIF-1α using immunohistochemistry. Moreover, mRNA expressions of ERK1, ERK2, Cylin D1 as well as HIF-1α in gastric mucosa were determined by quantitative real-time reverse transcription- polymerase chain reaction. RESULTS: We observed the appearance of active angiogenesis in GPL rats, and demonstrated that WPX could reduce microvascular abnormalities and attenuate early angiogenesis in most of GPL specimens with a concomitant regression of most intestinal metaplasia (IM) and a portion of gastric epithelial dysplasia (GED). In parallel, WPX could suppress HIF-1α mRNA expression (P < 0.01) as well as protein expression (although without statistical significance), and could markedly inhibit VEGF protein expression in GPL rats. Mechanistically, WPX intervention, especially at low dose, caused a significant decrease in the ERK1 and Cylin D1 mRNA levels. However, WPX might probably have no regulatory effect on ERK2 amplification. CONCLUSIONS: WPX could attenuate early angiogenesis and temper microvascular abnormalities in GPL rats. This might be partly achieved by inhibiting on the angiogenesis-associated markers HIF-1α and VEGF, and on the ERK1/Cylin D1 aberrant activation.

Asunto(s)

Inhibidores de la Angiogénesis/farmacología , Medicamentos Herbarios Chinos/farmacología , Mucosa Gástrica/efectos de los fármacos , Neovascularización Patológica , Neoplasias Gástricas/irrigación sanguínea , Estómago/efectos de los fármacos , Animales , Mucosa Gástrica/irrigación sanguínea , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patología , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Masculino , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , Ratas , Ratas Sprague-Dawley , Estómago/irrigación sanguínea , Estómago/patología , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patología , Factor A de Crecimiento Endotelial Vascular/metabolismo

RESUMEN

OBJECTIVE: To investigate the mechanism underlying the action of Weipixiao (WPX) in a rat's model with ameliorating gastric precancerous lesions (GPL). METHODS: HPLC analysis was performed to identify the chemical constituents of WPX preparation. Sprague- Dawley rats were randomly assigned into control group, model group, vitacoenzyme group, high-dose WPX group (H-WPX), medium-dose WPX group (M-WPX) and low-dose WPX group (L-WPX). After modeling, the treated rats were administrated WPX or vitacoenzyme intragastrically for consecutive 10 weeks. Gene and protein expressions of GSK3¦Â, C-myc, Cylin E were evaluated by quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR) and immunohistochemistry, respectively. RESULTS: WPX could efficiently attenuate the pathological alterations of ""non-progressive GPL"" in rats. As expected, mRNA and protein levels of C-myc and Cylin E were up-regulated in model rats, while GSK3¦Â expression down-regulated (P < 0.01). WPX treatment, especially at low dose, could significantly down-regulate the mRNA as well as protein levels of C-myc, and could lead to remarkable up-regulation of mRNA and protein levels of GSK3¦Â in GPL rats (P < 0.05). However, no significant changes were observed in WPX-treated rats. CONCLUSION: Our findings suggested that WPX-mediated attenuation of GPL pathological alterations might be due to its regulatory effect on the expressions of GSK3¦Â and C-myc, and on the dysregulation of Wnt/GSK3¦Â pathway.

Asunto(s)

Medicamentos Herbarios Chinos/administración & dosificación , Mucosa Gástrica/efectos de los fármacos , Glucógeno Sintasa Quinasa 3/metabolismo , Lesiones Precancerosas/tratamiento farmacológico , Lesiones Precancerosas/metabolismo , Proteínas Proto-Oncogénicas c-myb/metabolismo , Animales , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patología , Glucógeno Sintasa Quinasa 3/genética , Humanos , Masculino , Lesiones Precancerosas/genética , Lesiones Precancerosas/patología , Proteínas Proto-Oncogénicas c-myb/genética , Ratas , Ratas Sprague-Dawley

RESUMEN

Objective To investigate the clinical curative effect of Weipixiao for the treatment of chronic atrophic gastritis (CAG). Methods A total of 58 CAG patients with spleen-stomach deficiency syndrome was evenly randomized into Weipixiao group and Weifuchun group. Weipixiao group was treated with Weipixiao decoction orally,and Weifuchun group was treated with Weifuchun tablets orally. The treatment time covered 12 weeks. The scores of symptoms and pathology of CAG patients were graded before and after treatment. The clinical efficacy was evaluated with the symptom scores and pathological scores. Results (1)The scores of primary symptoms and secondary symptoms in the two groups were obviously decreased after treatment (P < 0.05 compared with those before treatment),and the decrease in Weipixiao group was superior to that in Weifuchun group(P< 0.05).(2) The total effective rate for clinical efficacy in Weipixiao group and Weifuchun group was 96.55%, 93.10%respectively, the difference being significant (P < 0.05). (3)After treatment, the scores of gastric mucosal atrophy and intestinal metaplasia in the two groups were obviously decreased after treatment (P < 0.05 compared with those before treatment),and the scores of gastric mucosal atypical hyperplasia were decreased obviously in Weipixiao group(P<0.05) but not obviously in Weifuchun group(P>0.05). The inter-group comparison results showed that Weipixiao group had better effect on decreasing gastric mucosal atrophy scores than Weifuchun group(P<0.05). Conclusion Weipixiao has good curative effect for the treatment of CAG patients with spleen-stomach deficiency syndrome through relieving the symptoms and improving the pathological changes of gastric mucosal atrophy and intestinal metaplasia.

RESUMEN

OBJECTIVE: To study the effects of Weipixiao (胃痞消, WPX) on Wnt pathway-associated proteins in gastric mucosal epithelial cells from rats with gastric precancerous lesions (GPL). METHODS: Sprague Dawley rats were randomly divided into control, model, vitacoenzyme (0.2 g·kg(-1)·day(-1)), WPX high-dose (H-WPX, 15 g·kg(-1)·day(-1)), WPX medium-dose (M-WPX, 7.5 g·kg(-1)·day(-1)) and WPX low-dose (L-WPX, 3.75 g·kg(-1)·day(-1)) groups. After successfully establishing the GPL model, the rats were consecutively administered WPX or vitacoenzyme by gastrogavage for 10 weeks. Differential expression of Leucine-rich repeat-containing G-proteincoupled receptor 5 (Lgr5), matrix metalloproteinase-7 (MMP-7), Wnt1, Wnt3a, and ß-catenin in gastric mucosal epithelial cells in all groups were immunohistochemically detected, and the images were taken and analyzed semiquantitatively by image pro plus 6.0 software. RESULTS: Gastric epithelium in the model group showed significantly higher expression levels of Lgr5, MMP-7, Wnt1, Wnt3a and ß-catenin than those of the control group(P<0.01). Interestingly, we also observed Lgr5+ cells, which generally located at the base of the gastric glandular unit, migrated to the luminal side of gastric epithelium with GPL. The expression levels of Lgr5, MMP-7, Wnt1, and ß-catenin were all down-regulated in the L-WPX group as compared with those of both model and vitacoenzyme groups (P<0.05). A similar, but nonsignificant down-regulation in expression level of Wnt3a was noted in all WPX groups (P>0.05). CONCLUSION: Our findings suggested that the therapeutic mechanisms of WPX in treating GPL might be related with its inhibitory effects on the expressions of Lgr5, MMP-7, Wnt1, ß-catenin and the aberrant activation of Wnt/ß-catenin pathway.

Asunto(s)

Medicamentos Herbarios Chinos/uso terapéutico , Células Epiteliales/metabolismo , Células Epiteliales/patología , Mucosa Gástrica/patología , Lesiones Precancerosas/tratamiento farmacológico , Neoplasias Gástricas/tratamiento farmacológico , Vía de Señalización Wnt/efectos de los fármacos , Animales , Medicamentos Herbarios Chinos/farmacología , Células Epiteliales/efectos de los fármacos , Inmunohistoquímica , Masculino , Metaloproteinasa 7 de la Matriz/metabolismo , Lesiones Precancerosas/patología , Ratas Sprague-Dawley , Receptores Acoplados a Proteínas G/metabolismo , Coloración y Etiquetado , Neoplasias Gástricas/patología , Proteínas Wnt/metabolismo , beta Catenina/metabolismo

RESUMEN

<p><b>OBJECTIVE</b>To study the effects of Weipixiao (胃痞消, WPX) on Wnt pathway-associated proteins in gastric mucosal epithelial cells from rats with gastric precancerous lesions (GPL).</p><p><b>METHODS</b>Sprague Dawley rats were randomly divided into control, model, vitacoenzyme (0.2 g·kg(-1)·day(-1)), WPX high-dose (H-WPX, 15 g·kg(-1)·day(-1)), WPX medium-dose (M-WPX, 7.5 g·kg(-1)·day(-1)) and WPX low-dose (L-WPX, 3.75 g·kg(-1)·day(-1)) groups. After successfully establishing the GPL model, the rats were consecutively administered WPX or vitacoenzyme by gastrogavage for 10 weeks. Differential expression of Leucine-rich repeat-containing G-proteincoupled receptor 5 (Lgr5), matrix metalloproteinase-7 (MMP-7), Wnt1, Wnt3a, and β-catenin in gastric mucosal epithelial cells in all groups were immunohistochemically detected, and the images were taken and analyzed semiquantitatively by image pro plus 6.0 software.</p><p><b>RESULTS</b>Gastric epithelium in the model group showed significantly higher expression levels of Lgr5, MMP-7, Wnt1, Wnt3a and β-catenin than those of the control group(P<0.01). Interestingly, we also observed Lgr5+ cells, which generally located at the base of the gastric glandular unit, migrated to the luminal side of gastric epithelium with GPL. The expression levels of Lgr5, MMP-7, Wnt1, and β-catenin were all down-regulated in the L-WPX group as compared with those of both model and vitacoenzyme groups (P<0.05). A similar, but nonsignificant down-regulation in expression level of Wnt3a was noted in all WPX groups (P>0.05).</p><p><b>CONCLUSION</b>Our findings suggested that the therapeutic mechanisms of WPX in treating GPL might be related with its inhibitory effects on the expressions of Lgr5, MMP-7, Wnt1, β-catenin and the aberrant activation of Wnt/β-catenin pathway.</p>

Asunto(s)

Animales , Masculino , Medicamentos Herbarios Chinos , Farmacología , Usos Terapéuticos , Células Epiteliales , Metabolismo , Patología , Mucosa Gástrica , Patología , Inmunohistoquímica , Metaloproteinasa 7 de la Matriz , Metabolismo , Lesiones Precancerosas , Quimioterapia , Patología , Ratas Sprague-Dawley , Receptores Acoplados a Proteínas G , Metabolismo , Coloración y Etiquetado , Neoplasias Gástricas , Quimioterapia , Patología , Proteínas Wnt , Metabolismo , Vía de Señalización Wnt , beta Catenina , Metabolismo

RESUMEN

Objective To observe the effects of Weipixiao on the ultrastructure of gastric mucosa capillaries in rats with precancerous lesions of gastric cancer ( PLGC). Methods The rats were randomized into six groups, including normal group, model group, Vatacoenayme Tablets group ( 0.2 g·kg-1·d-1) , and high-, middle-, and low- dose Weipixiao groups ( 15, 7.5, and 3.75 g·kg-1·d-1 respectively) . The rats received spontaneous intake of N-methyl-N’-nitro-nitrosoguanidine ( MNNG, 200 μg/mL) solution combined with irregular diet and intragastric administration of purgative herbs Xiao Chengqi Decoction ( 2 mL, containing 1 g/mL crude drug) for 18 weeks to induce spleen-deficiency PLGC. The pathological changes in gastric mucosa and the ultrastructure of gastric mucosa capillaries were observed under the transmission electron microscope. Results The model has been established successfully. Transmission electron microscopy results in the model group showed as severely swollen endothelial cells of gastric mucosa capillaries, severely-narrowing or even blocked vascular lumens, rough and discontinuous basement membrane, and swollen, degenerated or even absent pericytes. And the ultrastructure of gastric mucosa capillaries in high-, middle-, and low- dose Weipixiao groups were improved to some degrees, the effect of low-dose Weipixiao group being the best. Conclusion The improvement of the mucosal microcirculation of spleen-deficiency PLGC rats may be one of the pathohistological mechanisms of Weipixiao for spleen-deficiency PLGC.

RESUMEN

Objective To observe the effect of Weipixiao, a compound recipe which has the actions of strengthening spleen, resolving stasis and removing toxins, on the histopathological changes of gastric mucosal tissue in rats with gastric precancerous lesions ( GPL) . Methods SD rats were randomly divided into normal group, model group, Vitacoenzyme group (0.2 g·kg-1·d-1), and high-, middle-, and low-dose Weipixiao groups ( in the dose of 15, 7.5, 3.75 g·kg-1·d-1, respectively) . Except for the normal control group, the rats in other groups received spontaneous intake of N-methyl-N’-nitro-nitrosoguanidine ( MNNG) solution combined with irregular diet and oral use of purgative herbs for 18 weeks to induce GPL. From the 9th week, the mediation groups were simultaneously given corresponding medicine for 10 weeks. At the end of the experiment, the histopathological changes of gastric mucosal tissue in all groups were observed. Results Pathological scores of intestinal metaplasia and epithelial dysplasia in rat gastric mucosa of the model group were significantly increased ( P<0.01 compared with those of the normal group) , but were decreased in three Weipixiao groups to various degrees, particularly in low-dose Weipixiao group ( P<0.05 or P<0.01) . Conclusion Weipixiao can block and reverse gastric intestinal metaplasia and dysplasia in GPL rats to certain degrees, and low-dose Weipixiao may have better long-term effect for the prevention and treatment of GPL.