1.Effect of Dunhuang medical prescription Dabupi Decoction combined with oxaliplatin on inflammatory immunity of gastric cancer-bearing mice based on IL-17/NF-κB signaling pathway
Yuanding ZENG ; Yun SU ; Hongxia GONG ; Shiwei NIU ; Han ZHANG
Chinese Journal of Immunology 2024;40(1):92-96
Objective:To study anti-tumor effect of Dunhuang medical prescription Dabupi Decoction on gastric cancer-bearing mice and effect of Dabupi Decoction combined with oxaliplatin on inflammatory immunity of gastric cancer-bearing mice based on IL-17/NF-κB signaling pathway.Methods:Mouse model of MFC gastric cancer subcutaneously bearing tumor was established and ran-domly divided into model group,oxaliplatin group,high,medium and low doses of Dabupi Decoction combined with oxaliplatin groups[21.58,10.79,5.40 g/(kg·d)],with 10 mice in each group,male and female cage rearing.Administration began after 8 days of inoculation and continued for 14 days;next day after the last administration,eyeball blood was taken,mice were killed,tumor tissues were taken and weighed,and tumor inhibition rate was calculated.ELISA was used to detect contents of IL-17 and IL-6 in mice serum,immunohistochemistry(IHC),RT-qPCR and Western blot were used to detect IL-17,IL-6,NF-κB and p-NF-κB mRNA and protein expressions in mice tumor tissues,respectively.Results:Tumor inhibition rates of oxaliplatin group,high,medium and low doses of Dabupi decoction combined with oxaliplatin groups were 33.02%,52.92%,46.33%and 39.52%,respectively,and tumor quality of each treatment group was significantly lower than that of model group(P<0.01).High,medium and low doses of Dabupi Decoction combined with oxaliplatin groups were higher than that of oxaliplatin group.Compared with model group,contents of IL-17 and IL-6 in serum and expressions of IL-17,IL-6,NF-κB p65 and pNF-κB p65 mRNA and protein in tumor tissues in each treatment group were significantly reduced(P<0.01,P<0.05).Compared with oxaliplatin group,levels of IL-17 and IL-6 in serum and expres-sions of IL-17,IL-6,NF-κB p65 and pNF-κB p65 mRNA and protein in tumor tissues in high and medium doses of Dabupi Decoction combined with oxaliplatin groups were significantly reduced(P<0.01,P<0.05).Conclusion:Dunhuang medical prescription Dabupi Decoction has a certain anti-tumor effect on MFC gastric cancer-bearing mice,which can regulate inflammatory immunity and inhibit occurrence and development of gastric cancer by inhibiting IL-17/NF-κB signaling pathway.
2.Protective Effect of Huangqi Baihe Granules on Acute Brain Injury in Rats with High Altitude Hypoxia Based on HIF-1α/NF-κB/NLRP3 Signal Pathway
Yuanding ZENG ; Yun SU ; Hongxia GONG ; Wangjie CAO ; Yongqi LIU ; Yong HUANG ; Guangxian LENG ; Lixia LI
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(10):134-141
ObjectiveTo observe the effect of Huangqi Baihe granules on the hypoxia-inducible factor 1α (HIF-1α)/nuclear factor-κB (NF-κB)/NOD-like receptor hot protein domain related protein 3 (NLRP3) signaling pathway in a rat model of high altitude hypoxia. MethodSixty male SPF SD rats were randomly divided into blank group, model group, dexamethasone group (5 mg·kg-1), and high, middle, and low-dose groups of Huangqi Baihe granules (4.1, 2.05, 1.025 g·kg-1). Among them, each Chinese medicine group was administrated orally for continuously 14 d, once a day, and the dexamethasone group was injected intraperitoneally for continuously 3 d as the positive control group. On the 15th d, the model group, dexamethasone group, and high, middle, and low dose groups of Huangqi Baihe granules were exposed to the simulated high altitude, low pressure, and low oxygen environment in the animal low-pressure simulation cabin, and the exposure lasted for 3 d. Blood was collected from the abdominal aorta and serum was separated, and the brain tissue was taken after being killed. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in brain tissue. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in rat serum. Western blot was used to detect HIF-1α, NLRP3, phosphorylated nuclear factor-κB (p-NF-κB), NF-κB, desquamation D (GSDMD), and cysteine aspartate-specitis protein-1(Caspase-1) in rats of each group. The mRNA expression levels of HIF-1α, NLRP3, NF-κB p65, GSDMD, and Caspase-1 were detected by real-time quantitative polymerase chain reaction (Real-time PCR). ResultThe results of HE staining showed that as compared with the normal group, the pathological sections of brain tissues in the model group showed that pyramidal cells were loosely arranged and distributed in disorder, with different sizes. Compared with the model group, the pathological changes in pyramidal cells in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules were reduced. The results of ELISA showed that as compared with the normal group, the content of TNF-α, IL-6, and IL-1β in the serum of rats in the model group was significantly higher (P<0.01). Compared with the model group, the content of TNF-α, IL-6, and IL-1β in the serum of rats in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules decreased significantly (P<0.05, P<0.01). The results of Western blot showed that as compared with the normal group, the relative protein expression levels of HIF-1α, NLRP3, p-NF-κB p65, GSDMD, and Caspase-1 in the brain tissue of the model group were significantly higher (P<0.01). As compared with the model group, the relative expressions of HIF-1α, NLRP3, p-NF-κB p65, GSDMD, and Caspase-1 in the brain tissue of rats in the dexamethasone group and the high-dose group of Huangqi Baihe granules were significantly decreased (P<0.05, P<0.01). The relative protein expression levels of HIF-1α, NLRP3, p-NF-κB p65, and Caspase-1 in the brain tissue of rats in the middle-dose group of Huangqi Baihe granules decreased significantly (P<0.01), and the relative protein expression of HIF-1α in the brain tissue of rats in the low-dose group of Huangqi Baihe granules was reduced (P<0.05). The Real-time PCR analysis showed that as compared with the normal group, the mRNA expression levels of HIF-1α, NLRP3, NF-κB p65, GSDMD, and Caspase-1 in the brain tissue of the model group were significantly increased (P<0.01). As compared with the model group, the mRNA expression levels of HIF-1α, NLRP3, NF-κB p65, GSDMD, and Caspase-1 in the brain tissue of rats in the dexamethasone group were significantly decreased (P<0.01). The mRNA expression levels of HIF-1α, NF-κB p65, GSDMD, and Caspase-1 in the brain tissue of rats in the high-dose group of Huangqi Baihe granules decreased significantly (P<0.01). The mRNA expression levels of HIF-1α, NLRP3, and Caspase-1in the brain tissue of rats in the middle-dose group of Huangqi granules decreased (P<0.05, P<0.01). ConclusionThe protective effect of Huangqi Baihe granules on acute brain injury in low-pressure hypoxic rats may be related to the HIF-1α/NF-κB/NLRP3 signaling pathway.
3.Guiqi Baizhu Prescription Combined with Oxaliplatin Protects Intestinal Barrier of Tumor-bearing Mice with Gastric Cancer by Regulation of VIP/cAMP/PKA/AQP Signaling Pathway
Huancheng DONG ; Yun SU ; Hongxia GONG ; Wangjie CAO ; Jianzheng HE ; Yongqi LIU ; Han ZHANG ; Yuanding ZENG ; Congyi LI ; Qian KANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(5):129-136
ObjectiveTo investigate the protective effect of Guiqi Baizhu prescription combined with oxaliplatin on the intestinal barrier of tumor-bearing mice with gastric cancer by regulating downstream aquaporin 3 (AQP3) and aquaporin 4 (AQP4) through the vasoactive intestinal peptide (VIP)/cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling pathway. MethodThe gastric cancer cell lines MFC with a density of 1×107/mL were prepared into cell suspension. The tumor-bearing mouse model of gastric cancer was established by inoculating 0.2 mL cell suspension under the right axilla of mice. After successful modeling, mice were randomly divided into 5 groups, namely, model group, oxaliplatin group (10 mg·kg-1), and high, medium, and low-dose oxaliplatin + Guiqi Baizhu prescription groups (17.68, 8.84, 4.42 g·kg-1), with 10 mice in each group, and the remaining 10 mice were set as a blank group. Mice in each group were treated with Chinese medicine, oxaliplatin, or normal saline by gavage or intraperitoneal injection for 14 d. The next day after the last dose, blood was taken from the eyeball to separate serum and take colonic samples. Hematoxylin-eosin (HE) staining was used to observe the changes in tissue morphology. The content of D-lactate acid (D-LA) and diamine oxidase (DAO) in the serum was determined by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein expressions of VIP, cAMP, PKA, AQP3, and AQP4 were detected by Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. ResultCompared with the blank group, the model group showed edema in the colonic submucosa, disordered arrangement of intestinal glands in the mucosal layer, loss of goblet cells, infiltration of inflammatory cells, and villus shedding. However, there were different degrees of improvement in each administration group. As compared with the blank group, the serum levels of DAO and D-LA in the model group were significantly increased (P<0.01). As compared with the model group, the levels of DAO and D-LA in the high-dose oxaliplatin + Guiqi Baizhu prescription group and the level of D-LA in the medium-dose oxaliplatin + Guiqi Baizhu prescription group were decreased (P<0.05, P<0.01). As compared with the oxaliplatin group, the levels of D-LA in the high and medium-dose oxaliplatin + Guiqi Baizhu prescription groups were decreased (P<0.05), and the levels of DAO and D-LA in other administration groups were decreased as well, but the difference had no statistical significance. As compared with the blank group, the mRNA and protein expression levels of VIP, cAMP, PKA, AQP3, and AQP4 in the model group were significantly decreased (P<0.05, P<0.01). As compared with the model group, the mRNA and protein expression levels of VIP, cAMP, PKA, AQP3, and AQP4 in each administration group were increased, and those in the high-dose oxaliplatin + Guiqi Baizhu prescription group were significantly increased (P<0.05, P<0.01), while the protein expression level of cAMP in the medium-dose oxaliplatin + Guiqi Baizhu prescription group were increased (P<0.05). As compared with the oxaliplatin group, the protein expression levels of cAMP in the high-dose oxaliplatin + Guiqi Baizhu prescription group were increased (P<0.05), and the mRNA and protein expressions of these indexes in the other groups were also increased but the differences were not statistically significant. ConclusionGuiqi Baizhu prescription combined with oxaliplatin can regulate AQP3 and AQP4 through the VIP/cAMP/PKA signaling pathway to protect the intestinal barrier of tumor-bearing mice with gastric cancer.
4.Intervention Effect of Dabufei Decoction on Acute Lung Injury in Rats with High Altitude Hypoxia Based on HIF-1α/NLRP3 Signaling Pathway
Congyi LI ; Wangjie CAO ; Yong HUANG ; Yun SU ; Hongxia GONG ; Guangxian LENG ; Lixia LI ; Yuanding ZENG
Chinese Journal of Modern Applied Pharmacy 2024;41(6):736-742
OBJECTIVE
To investigate the intervention effect of Dabufei decoction on acute lung injury in rats with high altitude hypoxia by regulating the expression of the HIF-1α/NLRP3 signaling pathway and related molecules.
METHODS
Sixty SPF SD rats were randomly divided into blank group, model group, positive drug group, Dabufei decoction high-dose, medium-dose, and low-dose groups with 10 rats in each group. After 3 d of adaptation to feeding, the rats in the blank group and model group were given the same amount of normal saline by gavage, and the rats in Dabufei decoction high-dose, medium-dose, and low-dose groups were given gavage for 14 d, respectively. The positive drug group was given dexamethasone by intraperitoneal injection for three consecutive days before entering the chamber. Except for the blank group, the rats in each group were exposed to hypoxia in the experimental animal low-pressure simulation chamber from the 15th day for three consecutive days. At the end of the experiment, the wet to dry ratio(W/D) of the rat lung tissue was detected. The morphology of lung tissue was observed by HE staining. ELISA detected the levels of IL-1β and IL-18 in serum. Western blotting and RT-qPCR were used to detect the protein and mRNA expressions of HIF-1α, NLRP3, GSDMD, and caspase-1 in the lung tissue of rats.
RESULTS
The W/D value showed that compared with the blank group, the W/D of the model group was significantly increased(P<0.01). Compared with the model group, the W/D of rats in the positive drug group, Dabufei decoction high-dose group, medium-dose, and low-dose groups was significantly decreased(P<0.01 or P<0.05). HE results showed that compared with the blank group, alveolar septum thickening, pulmonary interstitial congestion, edema, inflammatory cell infiltration, and a small amount of exudation in the alveolar cavity were seen in the lung tissue of the model group. Compared with the model group, the thickening of alveolar walls in the positive drug group, Dabufei decoction high-dose group, medium-dose, and low-dose groups were reduced, and the pulmonary interstitial congestion, edema, and inflammatory cell infiltration were significantly reduced. ELISA results showed that IL-1β and IL-18 in rat serum were significantly higher in the model group than in the blank group(P<0.01). Compared with the model group, the levels of IL-1β and IL-18 in the serum of the rats in the positive drug group, Dabufei decoction high-dose group, medium-dose, and low-dose groups were significantly decreased(P<0.05 or P<0.01). Moreover, the results of Western blotting and RT-qPCR showed that compared with the blank group, the relative protein and mRNA expressions of HIF-1α, NLRP3, GSDMD, and caspase-1 in the lung tissue of the model group were significantly increased(P<0.01). Compared with the model group, the relative protein and mRNA of HIF-1α, NLRP3, caspase-1 and GSDMD in the positive drug group and Dabufei decoction high-dose group were significantly decreased(P<0.05 or P<0.01), the relative protein of HIF-1α, NLRP3, caspase-1 and GSDMD in Dabufei decoction medium-dose group were significantly decreased and HIF-1α, caspase-1 mRNA were significantly decreased(P<0.05), the relative protein of HIF-1α and GSDMD in the low-dose group was decreased(P<0.05). The positive drug group and Dabufei decoction high-dose group had the more significant effect.
CONCLUSION
Dabufei decoction can regulate the HIF-1α/NLRP3 signaling pathway, inhibit pyroptosis and reduce inflammation, and has a certain protective effect on acute lung injury in rats with high altitude hypoxia.