Histology and histopathology Vol.34, nº8 (2019)
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- PublicationOpen AccessTripterygium glycoside protects diabetic kidney disease mouse serum-induced podocyte injury by upregulating autophagy and downregulating β-arrestin-1(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Zhan, Huifang; Jin, Juan; Liang, Shikai; Zhao, Li; Gong, Jianguang; He, QiangBackground. Diabetic kidney disease (DKD), one of the most common causes of end-stage renal disease (ESRD), remains prevalent in many populations. Podocyte loss and apoptosis play a crucial role in the progression of DKD. Tripterygium glycoside (TG), a widely used Chinese herb, exerted comprehensive protective effects on preventing DKD progression. This study was performed to assess the podocyte protective effect of tripterygium glycoside on DKD by the potential role of activation of autophagy and downregulating βarrestin-1. Methods. Tripterygium glycoside and small interfering RNA (siRNA) of β-arrestin-1 were added to 10% db/db mice high-glucose serum induced podocytes in vitro. Autophagic activity was evaluated by transmission electronic microscopy, immunofluorescence staining and western blot analysis. Apoptotic activity was evaluated by Annexin V-FITC/PI flow cytometric analysis. The levels of nephrin and podocin, a marker protein of podocytes, were examined using western blot analysis. Results. Significantly ameliorated podocyte apoptosis, increased nephrin and podocin levels and inhibited expression of β-arrestin-1 were observed after pretreatment of tripterygium glycoside in DKD mouse serum treated podocytes. Significantly higher levels of autophagic activity were also observed. Silencing βarrestin-1 upregulated autophagic activity and ameliorated podocyte apoptosis. Silencing β-arrestin-1 in combination with tripterygium glycoside enhanced the levels of LC3-II and LC3-II/LC3-I ratios and reduced the expression of p62. Finally, we observed a notable reduction in podocyte apoptotic rate in DKD serum + siRNA-β-arrestin-1 + TG group compared to DKD serum + siRNA-β-arrestin-1 group, and upregulated protein levels of nephrin and podocin compared to treatment with siRNA-β-arrestin-1 only. Conclusions. This study demonstrated that tripterygium glycoside provided protection against podocyte injury induced by high-glucose serum, and that this effect was mediated by the concomitant activation of autophagy and downregulation of β-arrestin-1.
- PublicationOpen AccessHaptoglobin expression in human colorectal cancer(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Mariño Crespo, Óscar; Cuevas Álvarez, Elisa; Harding, Amy L.; Murdoch, Craig; Fernández Briera, Almudena; Gil Martín, EmilioAims and experimental design. The acutephase protein haptoglobin (Hp) has been recently detected in colorectal cancer (CRC) tissue, where its expression correlates with metastasis. Recently, we identified Hp as a CDw75 antigen-expressing protein in colorectal tissue. To deepen the knowledge of this protein in CRC, we studied the expression of Hp in healthy and tumour tissue specimens from 62 CRC patients by immunohistochemistry and Western blotting, as well as in the Caco-2 and HT-29 CRC cell lines by quantitative PCR, immunofluorescence microscopy and flow cytometry. Results and discussion. Hp immunopositive staining was absent in the 18 healthy colorectal specimens analysed, whereas it was observed in 24% (15/62) of the tumour specimens as cytoplasmic granules within cancer cells. Furthermore, Hp expression in CRC was associated with Dukes’ stage and the presence of metastasis in our population of study. In vitro cultured Caco-2 and HT-29 cells expressed mRNA for Hp and the protein was detected at the cell surface. Conclusions. This study confirms the expression of Hp in CRC, both in vivo and in vitro, and provides further evidence of its association with disease progression and metastasis.
- PublicationOpen AccessEccrine spiradenoma of the nipple: Case report, differential diagnosis and literature review(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Metovic, Jasna; Gallino, Chiara; Zanon, Eugenio; Bussone, Riccardo; Russo, Roberto; Vissio, Elena; Annaratone, Laura; Conti, Luca; Papotti, Mauro; Cassoni, Paola; Castellano, IsabellaEccrine spiradenoma is a rare lesion originating from eccrine sweat glands, with only few cases reported in breast tissue: we here describe for the first time, an eccrine spiradenoma arising in the nipple. An 84 year-old woman with a lesion enlarging her right nipple, showing ulcerations and eczema-like changes of the covering skin, was admitted to our hospital. Surgical excision of the central quadrant with nipple-areola complex was performed, followed by histopathological evaluation which revealed an adenoma with predominantly basaloid epithelial cells. The lesion was composed of tightly packed small and large groups of cells, arranged in diffuse alveolar/pseudorosette formations. The small cells expressed p63 and calponin, while a positive expression of CK7 and CD117 was detected in large cells. After careful and detailed examination, excluding various similar entities, a diagnosis of eccrine spiradenoma has been rendered. Although extremely rare, eccrine spiradenoma should be taken into account in the differential diagnosis of subcutaneous primary breast tumors.
- PublicationOpen AccessElevated blood/lymphatic vessel ratio in pterygium and its relationship with vascular endothelial growth factor (VEGF) distribution(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Martín López, Javier; Pérez Rico, Consuelo; García Honduvilla, Natalio; Buján, Julia; Pascual, GemmaIntroduction. Pterygium is a conjunctival fibrovascular tissue growth on the cornea. The pathogenesis of pterygium involves several factors such as the presence of active angiogenic factors. Expansion of the lymphatic microvasculature has also been hypothesized. This study examines the activity of the angiogenic/lymphangiogenic factor VEGF and the expression of vascular and lymphatic endothelial proteins in pterygia and normal conjunctival tissues. Materials and methods. Primary grade 2 pterygium (n=20) and normal conjunctiva (n=20) biopsies were obtained during surgery after written informed consent. mRNA expression for CD31, podoplanin, and VEGF (isoforms VEGF-A and VEGF-165) were determined by qRT-PCR. Tissue samples were also processed for immunohistochemical techniques to examine the lymphatic and vascular endothelium (anti-D2-40, antiCD31 respectively) and VEGF-A and VEGF-C levels and distribution. Results. VEGF-A gene expression levels failed to differ between the healthy and pterygium tissues. However, expression of its more angiogenic isoform, VEGF-165, was significantly higher in the pterygia. Immunohistochemistry revealed the greater presence of VEGF-A, compared to VEGF-C, in pterygium than conjunctiva, both in blood vessels and extracellular matrix. In addition, pterygia showed higher expression levels of the endothelial junction protein CD31. Lymphatic marker D2-40 expression was slightly augmented in this pathological tissue. The ratio between blood and lymphatic vessel counts was 1.05 in the normal conjunctiva and 3-fold this value in pterygium. Conclusion. In pterygium, while both lymphangiogenesis and angiogenesis take place, the formation of new blood vessels is the most relevant event, correlating with the increased expression of vascular endothelial CD31 and an elevated blood/lymphatic vessel ratio. The presence of high levels of VEGF-A in both vessel networks and extracellular matrix in human pterygium tissue may have a major impact on angiogenesis in this pathological tissue
- PublicationOpen AccessEpigallocatechin-3-gallate protects the testis from damage generated by experimental cryptorchidism in rabbits(Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Vigueras Villaseñor, Rosa María; Jiménez Cabrera, Tania; Chávez Saldaña, Margarita; Jiménez Trejo, Francisco; Cuevas Alpuche, Osvaldo; Rojas Castañeda, Julio CésarCryptorchidism (CO) is a risk factor for infertility in men. It is associated with an increase in oxidative stress which alters the differentiation of the gonocytes to spermatogonia. Epigallocatechin-3-gallate (EGCG) is an antioxidant that acts as a free radical scavenger and activates the antioxidant enzymes. The aim of this work was to investigate if EGCG plays a role in the protection of the testicle from alterations generated by CO and its possible mechanism. Male rabbits 7 days old were divided into four groups and distributed as follows: 1) control (C) treated with EGCG vehicle (V) (C/V); 2) C with administration of EGCG from 65 to 120 days postpartum (dpp) (C/EGCG); 3) CO induced by administration of 17β-estradiol plus EGCG vehicle (CO/V) and 4) CO plus EGCG administration (CO/EGCG). The animals were euthanized at 120 dpp and their testes were processed to evaluate lipid peroxidation, activities of superoxide dismutase (SOD) and catalase (CAT) enzymes as well as serum testosterone (T) concentrations. In addition, the rates of apoptosis, cell proliferation and histological alterations were determined. The CO/EGCG group showed a significant reduction in lipid peroxidation, a significant increase in the anti-oxidant enzyme activities and concentrations of T. Also, there was a significant decrease in the histological alterations, absence of gonocytes and active spermatogenesis when compared with CO/V group. These results show that EGCG reduces lipid peroxidation and increases the activity of the endogenous anti-oxidant system which protects the testes from alterations produced by oxidative stress generated during experimental CO.
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