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Objective This study aimed to establish a pre-metastatic niche mouse model utilizing luciferase-labeled Lewis (Luc-Lewis) lung cancer cells and to assess the efficacy of this model employing both qualitative and quantitative methods. Methods C57BL/6 mice were categorized into two groups: a normal control group and a model group, each containing 15 individual mice. The pre-metastatic niche model was established via tail vein injection of Luc-Lewis lung cancer cells. Body mass were measured daily for all groups. Tumor fluorescence signals within the mice were detected using a high-throughput enzyme marker instrument. Lung tissue specimens were harvested to evaluate metastatic progression. HE staining was used to assess histopathological changes. Real-time quantitative PCR and Western blot analysis were used to detect the mRNA and protein expression of lysyl oxidase (LOX), matrix metalloproteinase 9 (MMP9), versican (VCAN), and fibronectin (FN), which are the specific markers for the formation of the microenvironment of lung tissues before metastasis. Results Significant declines in body mass and observable lethargy were noted in the model group when compared to the control group. Distinct fluorescence signals were observed in the lung tissue of the model group, demonstrating a positive correlation with the duration of model establishment. By day 14, elevated mRNA and protein expression levels of LOX, MMP9, VCAN, and FN were significantly evident. In addition, histopathological evaluations revealed augmented interstitial thickness, alveolar atrophy and significant inflammatory cell infiltration within the lung tissues of the model group. By the 21st day, metastatic lesions manifested in the lung tissues of the model group, suggesting an approximate pre-metastatic niche maturation timeline of 14 days. Conclusion A pre-metastatic niche mouse model for Lewis lung cancer is successfully established.
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Ratones , Animales , Neoplasias Pulmonares/patología , Metaloproteinasa 9 de la Matriz , Ratones Endogámicos C57BL , Carcinoma Pulmonar de Lewis , Modelos Animales de Enfermedad , ARN Mensajero , Microambiente TumoralRESUMEN
Objective To investigate the effects of Astragalus polysaccharide (APS) on damage of keratinocytes in ultraviolet B (UVB) radiation skin in HaCaT cells; To preliminarily explore its mechanism of action.Methods HaCaT cells were treated by UVB irradiated for modeling. Blank control group, model group, UVB irradiation group and low-, medium- and high-dose APS interventional groups were set up. Each medication group was given relevant medicine. MTT assay was used to determine the cell activity; GSH-Px, CAT, SOD activity and MDA content were detected by biochemical colorimetric method; contents of TNF-α and IL-6 were detected by ELISA.Results Compared with the control group, the contents of SOD, CAT and GSH-Px in the model group decreased significantly, while the contents of MDA, TNF-α and IL-6 increased significantly (P<0.05); compared with the mode group, the contents of SOD, CAT and GSH-Px in the medication groups increased significantly, while the contents of MDA, TNF-α and IL-6 were reduced significantly (P<0.01).Conclusion APS can reduce the oxidative stress injury of UVB to HaCaT cells to some extent.
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Objective To establish a rat model of ulcerative colitis ( UC) with spleen and kidney Yang deficien-cy.Methods The rat model of ulcerative colitis with spleen and kidney Yang deficiency was established by oral adminis-tration of Rhubarb decoction, intramuscular injection of hydrocortisone, and combined with ethanol enema of TNBS (2,4, 6-trinitrobenzene sulfonic acid).Sixty Wistar rats ( body weight 180 ±20 g, male:female=1∶1) were randomly divided into blank control group and groups of UC models with spleen kidney Yang deficiency for 7 days, 14 days and 21days.The serum levels of FT3, FT4, and T of the rats were detected by ELISA assay.Results Compared with the blank control group, the serum levels of FT3, FT4, and T in the groups of UC rat models with spleen kidney Yang deficiency for 7 days, 14 days and 21days were decreased to a different extent (P<0.05), especially, decreased dramatically in the model group for 21 days.Conclusions FT3, FT4, and T are sensitive indexes with spleen and kidney Yang deficiency.The detection of serum levels of FT3, FT4, and T can better verify the spleen and kidney Yang deficiency in the rats, and prove that the spleen and kidney Yang deficiency type UC animal model is successfully prepared.
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BACKGROUND:Gastric cancer mesenchyal stem cel s from clinical stomach cancer specimens and tumorigenic tissues in nude mice are similar to the bone marrow mesenchymal stem cel s in biological characteristics, which have been proved to be an important component of tumor microenvironment to promote tumor growth. It is speculated that biological characteristic of bone marrow mesenchymal stem cel s may change in stomach cancer microenvironment. OBJECTIVE:To observe the effect of stomach cancer microenvironment on morphology and proliferation of bone marrow mesenchymal stem cel s and expressions of CD34 and CD44. METHODS:Rat bone marrow mesenchymal stem cel s were cultured alone as control group. In the test group, rat bone marrow mesenchymal stem cel s were co-cultured with human stomach cancer BGC-823 cel s using Transwel chamber assay to establish the stomach cancer microenvironment. Then, cel morphology, proliferation, cel cycle and CD34, CD44 expressions were observed and detected using inverted phase contrast microscope, MTT assay, and flow cytometry, respectively. RESULTS AND CONCLUSION:In the test group, bone marrow mesenchymal stem cel s were similar to human stomach cancer cel s BGC-823 that arranged disorderly and irregularly, were interconnected loosely, became thinner and longer, and grew in clusters with smal er nuclei. The cel proportion in G 1 phase significantly decreased, but that in S and G 2/M phases significantly increased (P<0.01, P<0.05). The positive rate of CD44 significantly declined, and the CD34 expression significantly raised (P<0.01). In conclusion, stomach cancer microenvironment by non-contact co-culture with BCG-823 cel s has an obvious effect on the morphology, proliferation and surface antigens expressions of bone marrow mesenchymal stem cel s that wil tend to be malignant gastric cancer cel s.
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Objective To detect the levels of IL-1, IL-6,TNF-αand IFN-γin serum and colon tissue of rat mod-els of ulcerative colitis with spleen and kidney Yang deficiency, and to explore their roles in the pathogenesis of ulcerative colitis ( UC) .Methods The rat model of ulcerative colitis with Yang deficiency of spleen and kidney was induced by perfusion of rhubarb decoction plus intramuscular injection of hydrocortisone and combined with TNBS (2,4,6-trinitro-benzenesulfonic acid) and ethanol enema.Sixty SPF wistar rats ( body weight 180 ±10 g, male:female=1:1) were ran-domly divided into blank control group, UC model with spleen kidney Yang deficiency for 7 days, 14 days and 21 days groups, respectively.The levels of IL-1, IL-6, TNF-αand IFN-γin serum and colon tissue were detected by ELISA.Re-sults Compared with the blank group, the levels of IL-1, IL-6, TNF-αand IFN-γin serum and colon tissue of rat UC model group with spleen kidney Yang deficiency were greatly increased (P<0.05), especially evidently increased in the model group at 21 days.Conclusions The pro-inflammatory cytokines IL-1, IL-6, TNF-αand IFN-γplay an important role in the pathogenesis of ulcerative colitis with syndrome of spleen and kidney Yang deficiency.
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Objective To detect mechanism of action mode of Jiuxieling Granules in spleen and kidney yang deficiency ulcerative colitis. Methods The perfusion of Rhei Radix et Rhizoma Decoction plus intramuscular injection of hydrocortisone and combined with TNBS and ethanol enema were employed to establish UC animal model. Ninety rats were randomly divided into blank group, model group, SASP group and Jiuxieling Granules 7 days, 14 days and 21 days groups. All treatment groups received relevant medicine intervention. The levels of IL-1, IL-6, TNF-α, and IFN-γin serum and colon tissue were detected by ELISA. Results Compared with the blank group, the levels of IL-1, IL-6, TNF-α, and IFN-γ in serum and colon tissues of rats in model group increased (P<0.05);compared with the model group, the levels of IL-1, IL-6, TNF-α, and IFN-γin serum and colon tissues of rats in treatment groups were reduced greatly (P<0.05), among which Jiuxieling Granules 21 days group showed the most obvious effects (P<0.05). Conclusion Jiuxieling Granules can regulate the normal secretion of the levels of IL-1, IL-6, TNF-α, and IFN-γin serum and colon tissue of model rats, and inhibit inflammation and protect colonic mucosa.
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Objective To observe the effects of Astragalus polysaccharide ( APS) on tumor growth, cytokine and immune function impairment induced by cisplatin ( DDP) in mice bearing Lewis lung cancer.Methods A total of 90 mice were used in this study:10 for blank control group, and 80 mice with transplanted Lewis lung cancer were randomly divided into 8 groups:model control group (physiological saline), positive control group treated with DDP (6 mg/kg), low dose APS (50 mg/kg), moderate dose APS (100 mg/kg) and high dose APS (200 mg/kg) groups and three combinations of APS+DDP groups ( the same three APS levels with half dose of DDP, respectively) .0.3 mL of the drugs was intraper-itoneally injected to the mice, respectively, on the second day after moldeling.DDP was injected once a week and other drugs were injected once per day for consecutive 20 days.On the 21st day, blood samples were collected and serum levels of cytokine IL-2, IL-6, IL-12 and TNF-αwere determined by ELISA, and the tumor inhibition rate and immune organ in-dexes were assessed.Results The tumor inhibition rates of the positive control, low, moderate and high dose APS groups and three combinations of APS+DDP groups of mice bearing Lewis lung carcinoma were 49.30%, 17.21%, 39.68%, 17.21%, 51.02%, 57.21%and 65.11%, respectively.Compared with the model group, P<0.05 or P<0.01, and compared the three combination groups with the DDP group, P<0.05.Compared with the blank control group, the spleen index was significantly increased in the moderate and high dose APS groups and the three combinations of APS +DDP groups.There was a significant difference between the spleen indexes of the model control group, and the spleen indexes of high dose APS and the combination with high dose APS groups were significantly higher than that of the model control group (P<0.05).Compared with the DDP group, APS in various doses and combinations increased the thymus index and spleen index.Conclusions APS can improve the levels of cytokine IL -2, IL-6, IL-12 and TNF-αin mice bearing Lewis lung cancer, enhance the immune function impairment induced by DDP, has certain protective effect on the immune organs, and inhibit the growth of Lewis lung cancer in mice.When APS is used in combination with a half-dose of DDP, APS enhanced the inhibition of tumor growth.This mechanism may be related to the enhanced body immune function.Our results indicate that APS enhances the therapeutic effect of DDP and reduces its toxicity, therefore, may have potential application value in future treatment of solid tumors.
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@#Objective To study the effect of electrical stimulation of vagus nerve on inflammatory response in septic shock rats. Methods SD rats were randomly divided into 5 groups: Group Ⅰ was the sham group, group Ⅱ with the cecal ligation and puncture (CLP) and the vagus nerve were isolated but not transected, group Ⅲ with bilateral cervical vagotomy following CLP, group Ⅳ with bilateral cervical vagotomy after CLP and the left vagus nerve trunks were stimulated with bipolar electrodes, group Ⅴ with bilateral cervical vagotomy after CLP and the right vagus nerve trunks were stimulated. The common carotid artery pressure was monitored, and the plasma tumor necrosis factor α (TNF-α), nitric oxide synthases (NOS) and nitric oxide (NO) were measured 2 h after stimulation. Results The mean arterial blood pressure (MAP) gradually decreased and the concentration of plasma TNF-α, NOS and NO significantly increased after CLP. Electrical stimulation of the left and right vagus nerve significantly increased the MAP and decreased the plasma TNF-α, NOS and NO levels. Conclusion Direct electrical stimulation of the left and right vagus nerve can significantly improve the blood pressure and reduced plasma TNF-α, NOS and NO levels during septic shock, which may play a role in anti-shock in rats.
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Objective To study the effects of single leaf Asarum on pulmonary ventilation in experimental rats with syndrome of cold fluid shooting lung. Methods Wistar rats were randomly divided into three groups including singe leaf Asarum group [fed with water extracts of singe leaf Asarum 1 mL/(kg?d)], singe leaf Asarumi Miq group [fed with water extracts of singe leaf Asarumi Miq 1 mL/(kg?d)] and normal saline group [fed with 0.9% normal saline 1 mL/(kg?d)]. After fed continually for 6 days, whole lung of rats were washed with normal saline [7 ℃, 2.5 mL/(kg?d)] and then were perfused for 30 seconds. This treatment was repeated 5 times every 5 minutes to make the model of cold fluid shooting lung. The fluid of perfusion was collected and detected. The changes of TXB2, ET, protein and phospholipids of the fluid were measured before and after lavage. Results TXB2, ET and protein in the perfusing fluid ascended obviously, while phospholipids were low in normal saline group. TXB2 and ET in singe leaf Asarum group and single leaf Asarumi Miq group were less than that in normal saline group (P
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OBJECTIVE: To study the influence of single leaf Asarum himalaicum on the renal function of rabbits. METHODS: Rabbits were divided into three groups. Asarum himalaicum, Asarum heterotropoides and normal saline were intravenously administered to the rabbits of one group respectively. The urine volume per minute, urine pH, urine glucose, protein and red blood cells, BUN, SCr, TXB2, 6-keto-PGF1alpha, TXB2/6-keto-PGF1alpha, endothelin, p-aminohippuric acid clearance rate and phenolsulfonphthalein excretion rate were tested before and after the administration. RESULTS: A certain dosage of single leaf Asarum himalaicum caused acute renal failure in rabbits. The indices tested were significantly different between rabbits administered Asarum himalaicum and normal saline. As compared with the rabbits administered Asarum heterotropoides, the results of indices tested decreased, but without statistical significance, except for SCr. CONCLUSION: The single leaf Asarum himalaicum can cause renal damage to rabbits. Its renal toxicity is lower that that of Asarum heterotropoides.