ABSTRACT
Objective:To investigate the efficacy of Balanophora involucrata Hook.f.in treatment of hyperuricemia(HUA)based on network pharmacology,molecular docking,and hyperuricemia models in vivo and in vitro,and to clarify the main targets of its active components and related signaling pathway mechanism.Methods:The potential targets of Balanophora involucrata Hook.f.in treatment of HUA were identified by Databases such as the Traditional Chinese Medicine Database in Taiwan,the Chinese Herbal Medicine Identification Database,Professional Chemical Database,TargetNet Database,SwissTargetPrediction Database,GeneCards,Therapeutic Target Database(TTD),DrugBank Database,DisGeNET Database,Online Mendelian Inheritance in Man(OMIM)Database,and Venny Database.STRING Database and Cytoscape software were used to construct the active component-predictive target network and protein-protein interaction(PPI)network for Balanophora involucrata Hook.f.;topological analysis was used to select the main active components and core targets;Gene Ontology(GO)functional and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were performed by R software;AutoDock Vina software was used for molecular docking validation.The NRK-52E cells were divided into blank control group,blank administration group,model group,and different concentrations(2.0,10.0,and 50.0 μmol·L-1)of erythrodiol(EDT)groups.High-performance liquid chromatography culture(HPLC)was used to detect the uric acid(UA)levels in the cell culture supernatants in various groups.The male ICR mice were divided into blank control group,blank administration group,model group,and EDT group;the mice in the last two groups were used to prepare the HUA models;kits were used to detect the levels of UA,creatinine(Cr),and blood urea nitrogen(BUN)in serum of the mice in various groups;the bilateral kidney tissue of the mice was harvested and weighed;the kidney indexes of the mice in various groups were calculated;TUNEL staining was used to observe the apoptosis in kidney tissue of the mice in various groups;Western blotting method was used to detect the expression levels of protein kinase B(AKT),phosphorylated AKT(p-AKT),phosphoinositide 3-kinase(PI3K),phosphorylated PI3K(p-PI3K),B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),and matrix metalloproteinase-9(MMP-9)proteins in kidney tissue of the mice in various groups.Results:Six active components of Balanophora involucrata Hook.f.were identified,involving 116 intersecting targets and 14 core targets.The enrichment analysis yielded 1 828 GO terms and 145 signaling pathways.The molecular docking results showed that EDT had good binding activity with MMP-9.The high uric acid cell experiment results showed that compared with blank control group,the UA level in the cells in model group was significantly increased(P<0.01);compared with model group,the UA levels in the cells in 2.0,10.0,and 50.0 μmol·L-1 EDT groups were significantly decreased(P<0.01).Compared with blank control group,the levels of UA,Cr,and BUN in serum of the mice in model group were increased(P<0.01),and the kidney indexes were significantly increased(P<0.01);compared with model group,the levels of UA,Cr,and BUN in serum of the mice in EDT group were decreased(P<0.05 or P<0.01),and the kidney index was significantly decreased(P<0.05 or P<0.01).Compared with blank control group,the number of apoptotic cells in kidney tissue of the mice in model group was increased;compared with model group,the number of the apoptotic cells in kidney tissue of the mice in EDT group was significantly decreased.Compared with blank control group,the ratios of p-AKT/AKT and p-PI3K/PI3K and expression level of Bcl-2 protein in kidney tissue of the mice in model group were significantly decreased(P<0.05 or P<0.01),while the expression levels of Bax and MMP-9 proteins were significantly increased(P<0.01);compared with model group,the ratios of p-AKT/AKT and p-PI3K/PI3K and expression level of Bcl-2 protein in kidney tissue of the mice in EDT group were significantly increased(P<0.05 or P<0.01),and the expression levels of Bax and MMP-9 proteins were significantly decreased(P<0.01).Conclusion:The active component of Balanophora involucrata Hook.f.,EDT,has a UA-decreasing effect and may inhibit the apoptosis and alleviate the kidney injury by activating the PI3K/AKT signaling pathway.
ABSTRACT
bcl-2 inhibitor of transcription 1 (Bit1) is an anoikis effector that can play a role of caspase-independent apoptosis by down-regulating bcl-2 expression. Previous studies have found that Bit1 affects cell survival and apoptosis through Erk and FAK-PI3K-Akt-NF-κB pathways. It is worth noting that the expression of Bit1 has shown the obvious tumor specificity in different tumors, and it is closely related to TNM stage, differentiation and prognosis of the tumor. This review summarizes the expression, main mechanism, and significance of Bit1 in different tumors.
ABSTRACT
Objective To investigate the beneficial role of synbiotics in the intestinal microbiota of patients with chronic functional constipation (CFC).Methods According to the inclusion and exclusion criteria,6 patients with CFC were enrolled with their fresh fecal samples collected,after a continuous treatment of one month their fresh fecal samples collected again.Meanwhile,6 healthy volunteers were enrolled as the control group with their fresh fecal samples collected.All samples were transported with ice and stored in -80 ℃ refrigerator,and were analyzed by metagenomics sequencing.Results After 4 weeks of symbiotic treatment,the intestinal microbiota had changed in species in patients with CFC.Bacteria of Escherichia_ coli,Clostridium_ sp._ SS2/1 and Clostridium_ sp._ 7_ 3_ 54FAA,which were rich in the people with constipation,decreased in abundance after the treatment.Bacteria of Lactobacillus_ oris and Bifidobacterium _ animalis,which were rich in the healthy people,increased in content after the treatment.Bacteria of Veillonella_ parvula,Veillonella_ sp._ 6_ 1_ 27,Veillonella_ sp._ 3 _ 1_ 44 which were rich in the healthy people,decreased in content after the treatment.LEfSe analysis showed that Parabacteroides distaso nis,Escherichia_ coli and Enterobacter-cloacae were the specific species of the three groups respectively.Conclusion Synbiotics can change the intestinal microbiota showing therapeutic effect,thus can be used as a novel clinical treatment method.
ABSTRACT
Objective To provide molecular genetic basis for oncobiological difference in left sided colon cancer and right sided colon cancer. Differentially expressed proteins in left sided colon cancer and right sided colon cancer were screened by proteomic technique. Methods Tissue samples including left sided colon cancer and right sided colon cancer were collected and preserved in the -80℃ refrigerator. In the first part of our experiment, protein was separated by 2-dimensional gel electrophoresis (2-DE) and the images of the gels were acquired by the scanner and then analyzed to find the differentially expression protein-spots in different groups. The peptide mass fingerprintings (PMF) was acquired by matrix assisted laser desorptiorn/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and the proteins were identified by data searching in the Mascotdatabase. Differentially expressed proteins were assayed by RT-PCR, Western blot, and immunohistochemical method. Results Altogether 55 differentially expressed protein spots were screened and 21 spots of them were identified. Compared with the right sided colon cancer, 14 proteins were up-regulated and 7 proteins down-regulated including HSP27 in the left sided colon cancer. HSP27 expressed higher in the right sided colon cancer than in the left sided colon cancer.Conclusion There are differentially expressed proteins in left sided colon cancer and right sided colon cancer, especially difference in HSP27 expression at mRNA and protein level, which may be molecular genetic basis for oncobiological difference in left sided colon cancer and right sided colon cancer.