ABSTRACT
To evaluate the therapeutic effect of Potentilla discolor on 2,4,6-trinitrobenzensulfonic acid(TNBS)-induced experimental ulcerative colitis(UC) in rats and to determine its therapeutic mechanism through mitochondrial autophagy, immune cells, and cytokines. A rat model of UC was established by TNBS-ethanol enema. Rats were divided into six groups: control, UC model, sulfasalazine(positive drug), and high-dose, moderate-dose, and low-dose ethanol extract groups. After 14-day continuous administration of the corresponding drugs, the disease activity index(DAI) and hematoxylin and eosin(HE) were evaluated. The morphological structure of mitochondria was observed by using transmission electron microscope(TEM), mitophagy-related mRNA expression was detected by using Real-time quantitative polymerase chain reaction(qRT-PCR), immune cell differentiation in rat serum was detected by using flow cytometry(FCM), and cytokine expression in colon tissues of rats was detected by protein microarray. The results showed that compared with the model group, each dose group of P. discolor could significantly reduce the DAI of UC model rats, and decrease the degree of inflammatory cells infiltration in the colon tissue of UC model rats. Meanwhile the expressions of T cells and Th cells in the serum increased significantly, the expression of Tc cells in the serum decreased significantly. Transmission electron microscope found that there was fusion of mitochondria and lysosomes in the colon tissue of the administration group. The expressions of mitochondrial autophagy related genes NF-κB, p62 and parkin were significantly increased in colon tissues. The results of protein chip showed that compared with the model group, the high dose group of P. discolor could significantly regulate the expression of cytokines. In conclusion, these results suggested that P. discolor improved TNBS-induced acute ulcerative colitis in rats by regulating the mitochondrial autophagy and the inflammatory factor expression.
Subject(s)
Animals , Rats , Autophagy , Colitis, Ulcerative/genetics , Colon , Mitochondria , Potentilla/geneticsABSTRACT
<p><b>OBJECTIVE</b>To identify the active material of anti-hepatic fibrosis from Amydae Carapax.</p><p><b>METHODS</b>Membrane separation technology was adopted to screen active fraction in Amydae Carapax, and the active components were isolated from the active fraction using gel chromatography and high performance liquid chromatography. The purified active components in Amydae Carapax were further analyzed using 4700 series time-of-flight mass spectrometer.</p><p><b>RESULTS</b>Proteins and peptides of Amydae Carapax with molecular weight less than 6000 were proved to have biological activity. 8 components (Bj1-Bj8) were isolated from the active fraction. Bj4, Bj6 and Bj7 were screened as active components. Bj7 was further purified, resulting in 7 components (Bj701-Bj707). Bj704 and Bj707 showed significant biological activity. Mass spectrometry showed three molecular ion peaks with highest abundance, i.e. m/e 526, 542 and 572, i.e. m/e 526, 542 and 572, in Bj707 -A The amino acid sequences of above three peptide compounds were NDDY (Asn-Asp-Asp-Tyr), NPNPT (Asn-Pro-Asn-Pro-Thr), and HGRFG (His-Gly-Arg-Phe-Gly), respectively. And M572 was the most abandunt components.</p><p><b>CONCLUSION</b>Three active peptide compounds of anti-hepatic fibrosis of Amydae Carapax were identified.</p>
Subject(s)
Animals , Humans , Cell Line , Liver Cirrhosis , Medicine, Chinese Traditional , Tissue Extracts , PharmacologyABSTRACT
Nasopharyngeal carcinoma (NPC) poses serious health problems in Southern China and yet the molecular mechanism of the carcinogenesis remains unclear. We used modern proteomic technologies to compare the protein expression profiles between the NPC cell lines (HNE1 and CNE1 ) and an immortalized nasopharyngeal epithelial cell line NP69 to identify cancer related proteins. Cell lysates were separated by two-dimensional gel electrophoresis (2 DE ) and analyzed by PDQuest software. The differentially expressed proteins were identified by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS). We discovered 15 up-regulated proteins and 18 down-regulated proteins in both HNE1 and CNE1 cell lines compared with NP69. These proteins are correlative with various functions, such as cell proliferation, apoptosis, cancer metastasis, metabolism, cytoskeleton and signal transduction. Western blotting analyses were further carried out to verify the differential expression of individual proteins. Several identified proteins in our research might be used as potential molecular markers to understand the molecular mechanism of NPC development and metastasis, and might be used as candidate targets for NPC treatments.
ABSTRACT
A subcellular proteomic method was applied to investigate the protein expression profiles of nasopharyngeal carcinoma (NPC) cell lines,CNE1 and CNE2,at various differentiation levels.Plasma membrane (PM) proteins were obtained by Percoll density grade centrifugation and subjected to twodimensional electrophoresis (2DE) followed by PDQuest software analysis.Nine proteins expressed with more than two folds difference were identified by MALDI-TOF-TOF,of which functions involved in cell differentiation,signal transduction,and metabolism.Half of these proteins,such as galectin-1 and annexin Ⅱ,were analyzed with real-time quantitative PCR or Westem blotting.We have tested a proteomic method to study differentiated NPCs at different levels and found several proteins that might be related to their biological characteristics.
ABSTRACT
<p><b>BACKGROUND</b>Pleomorphic hyalinizing angiectatic tumor (PHAT) of soft parts is a rare soft tissue tumor, which is generally considered low-grade. To distinguish the tumor from other soft tissue lesions, we analyzed the clinicopathologic and ultrastructural features, immunophenotypes, and flow cytometric DNA ploidy of PHAT in 9 cases.</p><p><b>METHODS</b>PHAT specimens were collected from 9 patients with PHAT from 1990 to 2004. Each specimen was cut into pieces and stained with hematoxylin-eosin, phosphotungstic acid-hematoxylin, Prussian blue, and Masson trichrome, respectively. Immunohistochemical stains for vimentin, S-100 protein, CD34, CD31, CD99, VEGF, desmin, CD117, alpha-SMA, and MIB-1 were performed with the Envision system. Flow cytometry was used in four specimens, two of which were observed by electron microscopy.</p><p><b>RESULTS</b>In the 9 cases, the PHAT occurred at the lower extremity in 2 patients, inguinal in 2, waist in 1, forearm in 1, buttock in 1, foot in 1, and the chest wall in 1. All the lesions presented in the superficial subcutaneous tissues. Follow-up data were available in 7 of the patients, among whom 2 (28.6%) had recurrence after primary therapy. Microscopically, typical PHAT was characterized by sheet-like proliferation of spindle or pleomorphic cells and clusters of thin-walled hyalinized cstatic vessels. In some areas of the tumor, hemosiderin-laden spindle cells, numerous small single vessels, and myxoid extracellular matrix could be identified, indicating an "atypical PHAT". Mitotic figures were rare in all the cases. In 5 of the 9 patients (55.6%), the tumor was typical PHAT; and in the other 4 (44.4%), typical and atypical PHAT coexisted. Immunohistochemically, the neoplastic cells were positive for vimentin, CD34, CD99, and VEGF, but negative for S-100 protein, desmin, SMA, and CD31. In all the cases, the MIB-1 proliferative activity of the neoplastic cells was lower than 2%. Ultrastructural analysis did not reveal any evidence of specific differentiation. Aneuploidy was not detected by flow cytometry.</p><p><b>CONCLUSIONS</b>Histologically, typical PHAT is characterized by spindle and pleomorphic cells associated with an angiectatic vasculature. The neoplastic cells often express vimentin and CD34, and may be positive for CD99 and VEGF. Ultrastructurally, the tumor usually has no specific differentiation. The low MIB-1 index and the absence of aneuploidy in PHAT indicate a non-malignancy. However, we consider the tumor as a borderline neoplasm because of its aggressive behaviour, and suggest wide local resection with tumor-free margin for the treatment of the disease.</p>