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Aim To observe the effect of Gupi Xiaoji Decoction (GPXJY) on the structure and function of mitochondria of human hepatoma cell HepG2 cells and explore its possible mechanism. Methods CCK8 was used to detect cell proliferation, Mito-Tracker Green fluorescence staining was used to observe the mitochondrial structure, flow cytometry was used to detect the membrane potential, Elisa was used to detect the ATP content, fluoroscopic electron microscopy was used to observe the microstructure changes, and high-content screening(HCS) was used to detect the related proteins. Results Fluorescence staining showed that GPXJY damaged the mitochondria of HepG2 cells and decreased the content of ATP. The results of flow cytometry showed that GPXJY could reduce the mitochondrial membrane potential of HepG2 cells. The results of electron microscope showed that GPXJY made the mitochondria of cancer cells swell and so on. HCS found that GPXJY significantly reduced the average fluorescence intensity of Bcl-2 in HepG2 cells, and significantly increased the average fluorescence intensity of apoptosis promoting proteins Bax, cytochrome-c, caspase-3 and cleaved-caspase-3, which was statistically significant. Conclusion GPXJY can regulate the structure and function of mitochondria in HepG2 cells.
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ObjectiveTo screen the active antitumor components of Gupi Xiaoji decoction by network pharmacology and molecular docking based on the pyroptosis mediated by cysteinyl aspartate-specific protease 1 （Caspase-1） and explore its molecular mechanism in intervening in the pyroptosis of HepG2.2.15 cells through in vitro experiments. MethodThe compounds and targets of Gupi Xiaoji decoction were screened out by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） to obtain the corresponding gene symbols. The targets of Caspase-1 were collected from GeneCards，online mendelian inheritance in man（OMIM），PharmGKB，and TTD，and the compound-gene target regulatory network was constructed by Cytoscape. The protein-protein interaction（PPI） network was established and analyzed by STRING. The mechanism of the effective components of Gupi Xiaoji decoction on Caspase-1 was predicted by gene ontology（GO） functional enrichment and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analyses. The molecular docking was verified with AutoDock Vina. The plasma medicated with Gupi Xiaoji Decoction was prepared and HepG2.2.15 cells were cultured in vitro. HepG2.2.15 cells were divided into a blank plasma group，a VX-765 group，a VX-765+medicated plasma group， and a medicated plasma group. After 48 hours of intervention with 15% medicated plasma， the expression and distribution of gasdermin D-N （GSDMD-N） on the surface of the cell membrane were detected by immunofluorescence staining. The release of lactic dehydrogenase （LDH）， interleukin（IL）-1β，and IL-18 in the cell supernatant was measured by enzyme-linked immunosorbent assay（ELISA） kits. The expression of Caspase-1 and GSDMD-N was measured by Western blot. ResultThe mitogen-activated protein kinase 14 （MAPK14），MAPK1，protein kinase B1 （Akt1）， MAPK8， V-Jun sarcoma virus oncogene homolog （JUN）， and TP53 screened by network pharmacology were the main targets. The compounds 7-hydroxy-5，8-dimethoxy-2-phenyl-chromone，wogonin，rhamnazin，moslosooflavone，isorhamnetin，7-O-methylisomucronulatol，formononetin，calycosin，luteolin，quercetin，kaempferol，β-sitosterol，and baicalein screened by network pharmacology were the main active components of Gupi Xiaoji decoction. Go enrichment analysis showed that multiple biological processes were involved， including responses to oxidative stress and metal ions，ubiquitin-like protein ligase binding，and phosphatase binding. KEGG pathway enrichment analysis showed MAPK pathway，nuclear factor（NF）-κB pathway，p53 pathway， and hypoxia-inducible factor-1（HIF-1） pathway were involved. Molecular docking showed that the targets had good binding with the components. In vitro experiments displayed that compared with the blank plasma group，the VX-765 group showed weakened GSDMD-N fluorescence signal，reduced release of LDH，IL-1β，and IL-18，and declining expression of Caspase-1 and GSDMD-N（P<0.01）， and the medicated plasma group showed increased GSDMD-N fluorescence signal， increased release of LDH，IL-1β，and IL-18，and up-regulated expression of Caspase-1 and GSDMD-N（P<0.01）. ConclusionGupi Xiaoji Decoction can induce the pyroptosis of HepG2.2.15 cells by regulating Caspase-1 through multiple targets and multiple pathways.
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@#Objective: To investigate the effects of XIAOJI Decoction combined with FOLFOX chemotherapy on serum cytokine expression profile in patients with advanced colorectal carcinoma by liquid chip technology. Methods: Fourteen patients with advanced colorectal carcinoma, who met the inclusion criteria and were treated in the Department of Oncology, Higher Education Mega Center Hospital of Guangdong Provincial Hospital of Traditional Chinese Medicine during January 1, 2018 and December 31, 2018 were retrospectively analyzed in this study. The patients were divided into chemotherapy group (n=7, treated with 5-Fluorouracil + Calcium Folic Acid+Oxaliplatin (FOLFOX)) and combined treatment group (n=7, treated with XIAOJI Decoction + FOLFOX) according to therapeutic scheme. The curative efficacy was evaluated after 6 treatment courses. The expression profile of cytokines in blood serum of patients was examined by liquid chip technology after every 2 courses. Results: Fourteen patients received a total of 84 cycles of therapy. Survival analyses showed that the progress-free survival time (PFS) and overall survival time (OS) of two groups couldn't be compared due to insufficient samples, although the combined treatment group had longer PFS (10 months vs 6 months) and OS (17 months vs 12 months) than the chemotherapy group.As to adverse reactions, the rates of leucopenia, diarrhea, nausea, peripheral neuritis and alopecia in two groups were comparable, while the severity in combined treatment group were lighter than that in chemotherapy group. In comparison with the combined treatment group, concentrations of serum BDNF and IL-2 were statistically higher in the chemotherapy group (P<0.05). By comparing the cytokine concentrations at different collection time points before and after the treatment, it showed that the concentration of serum IL-2 in chemotherapy group was higher than that in combined treatment group after 2 courses of treatment (P<0.05). In total, there were 19 cytokines showed a tendency to be higher in combined treatment group than chemotherapy group during different treatment periods. Conclusion: Combined treatment of XIAOJI Decoction with FOLFOX for advanced colorectal carcinoma is a treatment option worth exploring, and liquid chip analysis showed that the mechanism may be related to the reduction of serum LI-2 and BDNF levels in patients.
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Objective To investigate the apoptosis-inducing effect of the Tongbu-Xiaoji decoction on A549 cells.Methods A549 cell lines were cultured with normal serum and different concentration of serum containing Tongbu-Xiaoji decoction.The cells were stained with PI and Annexin V,and were evaluated by flowcytometry.Resnlts A549 cells were treated by Tongbu-Xiaoji decoction with low、median and high dose medicated serum,the rate of apoptosis in 24h were (5.92±2.04)％,(12.57±1.96)％,and (16.51±1.52)％respectively; the rate of apoptosis in 48h were (13.15±1.60)％,(26.42±2.30)％,and (31.47±2.08)％respectively and the rate of apoptosis in 72 h were (18.73 ± 1.89) ％,(31.72± 1.99) ％,and (36.92± 1.38) ％respectively,all were lower than compared with the model control group in 24 h were (0.40±0.08)％,48 h were (0.58± 0.16)％,72 h were (0.85 ± 0.13)％.Conclusion Tongbu-Xiaoji decoction has the apoptosis-inducing effect on A549 cells.
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ive] To observe the therapeutic effects of Xiaoji Decoction (XD) in treating intermed iate and late stages of lung cancer. [ Methods] One hundred and twelve patients with lung cancer in stage ? -? were randomly allocated to Group A, Group B and Group C. Group A ( n = 49) was treated with XD alone, Group B ( n = 33 ) was treated with various chemotherapeutic regimens according to the histo-logical types of lung cancer: CAP regimen for lung squamous carcinoma, EP regimen for lung adenocarci-noma and CE regimen for small cell carcinoma of lung, In Group C, 30 patients were treated with chemo therapy combined with XD. Symptomatic relief, life quality, tumor size, distant metastasis, sub-types of T lymphocytes, survival period and side effects in the three groups were evaluated after one course of treatment. [Results] The effects of XD on tumor size were evaluated by the effective rate and stabilized rate. They were 4.08%and 53.06%, 21.21% and 48.48%, and 46.67%and 76.67%in Group A, Group B and Group C respectively. The differences between Group A and Group B were significant ( P
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Purpose:To study the effect of the herbal decoction Qingyi Xiaoji Formula(QYXJ) on the proliferation of human pancreatic cancer cell line SW1990 in vivo and to explore the mechanism of its functions by means of cDNA microarray.Methods:Tumor-burdened nude mice were randomized into control group,5-FU group,and QYXJ groups at different dosages.After treatment,inhibiting rates of tumor growth were calculated.Tumor mRNA of the control group and the QYXJ group at moderate dose was extracted.The fluorescent cDNA probes were prepared,labeled with two different dyes Cy3 and Cy5,and then hybridized with cDNA microarray and scanned for fluorescent intensity.The genes with different expression were identified through the analysis of gene expression profile.Results:Inhibition rates of tumor growth in the QYXJ groups were 21.31%,38.16% and 29.09%,in the dose of 18 g/kg,36 g/kg,and 72 g/kg respectively.7 genes with reduced expression were identified,the functions of which were oncogene,protein translation and synthesis,DNA synthesis and repair,cell signal transduction,etc.Conclusions:QYXJ decoction may inhibit the proliferation of pancreatic cancer in vivo,possibly by blocking the action of an oncogene and its downstream signaling,or by regulation of protein synthesis in cancer cells.