Histology and histopathology Vol.35,nº12 (2020)

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Open Access
No rapid and demarcating astroglial reaction to stab wounds in Agama and Gecko lizards and the caiman Paleosuchus - it is confined to birds and mammals
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Lőrincz, Dávid; Kálmán, Mihály
y. The present study proves that the rapid and demarcating astroglial reactions are confined to birds and mammals. To understand the function of post-lesion astroglial reaction, the phylogenetical aspects are also to be investigated. Considering the regenerative capabilities, reptiles represent an intermediate position between the brain regeneration-permissive fishes and amphibians and the almost non-permissive birds and mammals. Damage is followed by a rapid astroglial reaction in the mammalian and avian brain, which is held as an impediment of regeneration. In other vertebrates the reactions were usually observed following long survival periods together with signs of regeneration, therefore they can be regarded as concomitant phenomena of regeneration. The present study applies short post-lesion periods comparable to those seen in mammals and birds for astroglial reactions. Two species of lizards were used: gecko (leopard gecko, Eublepharis macularius, Blyth, 1854) and agama (bearded dragon, Pogona vitticeps, Ahl, 1926). The gecko brain is rich in GFAP whereas the agama brain is quite poor in this. Crocodilia, the closest extant relatives of birds were represented in this study by Cuvier's dwarf caiman (Paleosuchus palpebrosus, Cuvier, 1807). The post-lesion astroglial reactions of crocodilians have never been investigated. The injuries were stab wounds in the telencephalon. The survival periods lasted 3, 7, 10 or 14 days. Immunoperoxidase reactions were performed applying anti-GFAP, anti-vimentin and anti-nestin reagents. No rapid and demarcating astroglial reaction resembling that of mammalian or avian brains was found. Alterations of the perivascular immunoreactivities of laminin and β-dystroglycan as indicators of glio-vascular decoupling proved that the lesions were effective on astroglia. The capability of rapid and demarcating astroglial reaction seems to be confined to mammals and birds and to appear by separate, parallel evolution in them.
Open Access
Sympathetic nervous system contributes to orthodontic tooth movement by central neural regulation from hypothalamus
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Cao, Haifeng; Fang, Bing; Wang, Xudong; Zhou, Yanheng
Orthodontic tooth movement (OTM) is a specific treatment of malocclusion, whose regulation mechanism is still not clear. This study aimed to reveal the relationship between the sympathetic nervous system (SNS) and OTM through the construction of an OTM rat model through the utilization of orthodontic nickeltitanium coiled springs. The results indicated that the stimulation of SNS by dopamine significantly promote the OTM process represented by the much larger distance between the first and second molar compared with mere exertion of orthodontic force. Superior cervical ganglionectomy (SCGx) can alleviate this promotion effect, further proving the role of SNS in the process of OTM. Subsequently, the ability of orthodontic force to stimulate the center of the SNS was visualized by the tyrosin hydroxylase (TH) staining of neurons in ventromedial hypothalamic nucleus (VMH) and arcuate nucleus (ARC) of the hypothalamus, as well as the up-regulated expression of norepinephrine in local alveolar bone. Moreover, we also elucidated that the stimulation of SNS can promote osteoclast differentiation in periodontal ligament cells (PDLCs) and bone marrow-derived cells (BMCs) through regulation of receptor activator of nuclear factor-κB ligand (RANKL)/osteoprotegerin (OPG) system, thus promoting the OTM process. In conclusion, this study provided the first evidence for the involvement of the hypothalamus in the promotion effect of SNS on OTM. This work could provide a novel theoretical and experimental basis for further understanding of the molecular mechanism of OTM.
Open Access
Farnesoid X receptor: a potential therapeutic target in multiple organs
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Zhang, Chao; Wang, Zixuan; Feng, Qingqing; Chen, Wei-Dong; Wang, Yan-Dong
Farnesoid X receptor (FXR), a member of the nuclear receptor family, is a common receptor found in the intestine and liver, and helps to maintain systemic metabolic homeostasis through regulating bile acid, glucose, lipid metabolism, and energy homeostatsis. In addition, FXR regulates the functions of various organs, such as liver, intestine, kidney, breast, pancreas, cardiovascular system and brain. FXR also plays a key role in regulation of gut-microbiota through mediating the various signaling pathways. Accordingly, FXR has become an attractive therapeutic target in a variety of diseases. This review combines classical and recent research reports to introduce the basic information about FXR and its important roles in various organs of the body.
Open Access
CD9 expression in vascular aging and atherosclerosis
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Kim, Jae-Ryong; Cho, Joon Hyuk
CD9 is a transmembrane glycoprotein belonging to the tetraspanin family. CD9 expression has been reported to be associated with cellular signaling, cell adhesion, cell migration, and tumor related processes. The aim of this study was to examine the immunohistochemical expression of CD9 in vascular senescence and atherosclerosis. One hundred and twenty samples of normal young arteries (obtained from individuals aged 0-60 years), 40 samples of normal old arteries (obtained from individuals aged 61-80 years), and 67 samples of atherosclerotic arteries were obtained from surgically resected specimens. Tissue microarray blocks were prepared for immunohistochemical staining. Immunohistochemical staining detected CD9 expression in 10.8% (13 of 120 samples) of normal young arteries and 30.0% (12 of 40 samples) of normal old arteries. CD9 expression was absent or mildly present in the smooth muscle cells and endothelial cells of normal arteries. Normal old arteries showed significantly higher expression of CD9 than normal young arteries (P<0.01). Atherosclerotic arteries showed moderate or strong CD9 expression (65 of 67 samples, 97.0%), which was observed in the smooth muscle cells, endothelial cells, macrophages, and atheromatous plaques. CD9 was significantly expressed in the atherosclerotic arteries compared to normal young and old arteries (P<0.01). The results suggest that CD9 expression may play an important role in the vascular senescence and pathogenesis of atherosclerosis.
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Nephrotoxicity of cypermethrin in rats. Histopathological aspects
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Alalwani, Aisha D.
Cypermethrin (CYP) is an important type II pyrethroid pesticide widely used to protect crops against pests and insect infestations. However, its toxicity is a risk to both human health and the surrounding environment. The present study was conducted to investigate the nephrotoxic effect and histopathological changes caused by cypermethrin in the kidney tissues of adult wistar rats. In this study, 30 Wistar rats were equally divided into three groups. G1, control animals; G2 and G3 treated with various sub lethal doses of CYP for 30 days as follows: G2, administered low dose (1/100 of LD50) of CYP; G3, administered high dose (1/50 of LD50) of CYP. The damage to different organelles of renal proximal and distal cells was observed using transmission electron microscopy. Histopathological damage in kidney samples was confirmed using morphological and histological measures. The results showed that CYP caused significant histopathological damage to the renal proximal and distal tubules of treated rats. Compared to control samples, CYP caused marked alterations in the dimensions of nucleus, ovoid and filamentous mitochondria of the treated cells. In conclusion, cypermethrin is found to be toxic to mammals. It caused marked ultrastructural damage to the renal proximal and distal tubules of wistar rats and the intensity of nephrotoxicity correlated with the dose of oral administration.
Open Access
Proliferative and chondrogenic potential of mesenchymal stromal cells from pluripotent and bone marrow cells
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Sfougataki, Irene; Varela, Ioanna; Stefanaki, Kalliope; Karagiannido, Angeliki; Roubelakis, Maria G.; Kalodimou, Vasiliki; Papathanasiou, Ioanna; Traeger-Synodinos, Joanne; Kitsiou-Tzeli, Sofia; Kitra, Vasiliki; Tsezou, Aspasia; Maria, Tzetis; Goussetis, Evgenios
Introduction. Mesenchymal stromal cells (MSCs) can be derived from a wide range of fetal and adult sources including pluripotent stem cells (PSCs). The properties of PSC-derived MSCs need to be fully characterized, in order to evaluate the feasibility of their use in clinical applications. PSC-MSC proliferation and differentiation potential in comparison with bone marrow (BM)-MSCs is still under investigation. The objective of this study was to determine the proliferative and chondrogenic capabilities of both human induced pluripotent stem cell (hiPSC-) and embryonic stem cell (hESC-) derived MSCs, by comparing them with BMMSCs. Methods. MSCs were derived from two hiPSC lines (hiPSC-MSCs), the well characterized Hues9 hESC line (hESC-MSCs) and BM from two healthy donors (BMMSCs). Proliferation potential was investigated using appropriate culture conditions, with serial passaging, until cells entered into senescence. Differentiation potential to cartilage was examined after in vitro chondrogenic culture conditions. Results. BM-MSCs revealed a fold expansion of 1.18x105 and 2.3x105 while the two hiPSC-MSC lines and hESC-MSC showed 5.88x1010, 3.49x108 and 2.88x108, respectively. Under chondrogenic conditions, all MSC lines showed a degree of chondrogenesis. However, when we examined the formed chondrocyte micromasses by histological analysis of the cartilage morphology and immunohistochemistry for the chondrocyte specific markers Sox9 and Collagen II, we observed that PSC-derived MSC lines had formed pink rather than hyaline cartilage, in contrast to BM-MSCs. Conclusion. In conclusion, MSCs derived from both hESCs and hiPSCs had superior proliferative capacity compared to BM-MSCs, but they were inefficient in their ability to form hyaline cartilage
Open Access
Glucocorticoid receptor modulates dendritic cell function in ulcerative colitis
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Yang, Xinxin; Geng, Jingshu
Ulcerative colitis (UC) is a serious form of inflammatory bowel disease (IBD) occurring worldwide. Although anti-TNF therapy is found to be effective in over 70% of patients with UC, nearly one-third are still deprived of effective treatment. Because glucocorticoids (GC) can effectively inhibit granulocyte-recruitment into the mucosa, cytokine secretion and T cell activation, they are used widely in the treatment of UC. However, remission is observed in only 55% of the patients after one year of steroid use due to a condition known as steroid response. Additionally, it has been noted that 20%-40% of the patients with UC do not respond to GC treatment. Researchers have revealed that the number of dendritic cells (DCs) in patients with UC tends to increase in the colonic mucosa. Many studies have determined that the removal of peripheral DCs through the adsorption and separation of granulocytes and monocytes could improve tolerance of the intestine to its symbiotic flora. Based on these results, further insights regarding the beneficial effects of Adacolumn apheresis in patients subjected to this treatment could be revealed. GC can effectively inhibit the activation of DCs by reducing the levels of major histocompatibility complex class II (MHC II) molecules, which is critical for controlling the recruitment of granulocytes. Therefore, alternative biological and new individualized therapies based on these approaches need to be evaluated to counter UC. In this review, progress in research associated with the regulatory effect of glucocorticoid receptors on DCs under conditions of UC is discussed, thus providing insights and identifying potential targets which could be employed in the treatment strategies against UC
Open Access
Cadherins down-regulation: towards a better understanding of their relevance in colorectal cancer
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Losi, Lorena; Zanocco-Marani, Tommaso; Grande, Alexis
The down-regulation of cadherin expression in colorectal cancer (CRC) has been widely studied. However, existing data on cadherin expression are highly variable and its relevance to CRC development has not been completely established. This review examines published studies on cadherins whose downregulation has been already demonstrated in CRC, trying to establish a relationship with promoter methylation, the capacity to influence the Wnt / CTNNB1 (catenin beta 1, beta-catenin) signaling pathway and the clinical implications for disease outcome. Moreover, it also analyses factors that may explain data variability and highlights the importance of considering the altered subcellular localization of the examined cadherins. The results of this survey reveal that thirty of one hundred existing cadherins appear to be down-regulated in CRC. Among these, ten are cadherins, sixteen are protocadherins, equally divided between clustered and non clustered, and four are cadherin - related. These findings suggest that, to better define the role played by cadherin down-regulation in CRC pathogenesis, the expression of multiple rather than individual cadherins should be taken into account and further functional studies are necessary to clarify the relative ability of individual cadherins to inhibit CTNNB1 therefore acting as tumor suppressors.
Open Access
Tripartite motif-containing 35 (TRIM35) is up-regulated in UUO-induced renal fibrosis animal model
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Chen, Yu; Ding, Yue; Wang, Li-Ming
Renal fibrosis has been recognized as a serious health threat in the world because of the high cost of treatment and poor prognosis. However, the molecular mechanism of renal fibrosis is still largely unknown. In this study, we aimed at illustrating the role of TRIM35 in the renal fibrosis process. A UUO mouse model and a TGF-β1-induced tubulointerstitial fibrosis model were constructed for the research of renal fibrosis at animal and cell level, respectively. Hematoxylin-eosin and Masson staining were used for visualizing the pathological change. qRT-PCR, Western blot analysis and immunohistochemical staining were used to detect the expression of fibrosis-associated proteins and TRIM35. The results showed that, after the modeling, the expressions of α-SMA, Collagen I, Collagen III, Fibronectin and Snail1 were up-regulated, while the expression of E-cadherin was down-regulated, indicating the successful construction of animal and cell models. More importantly, TRIM35 was proved to be upregulated in both animal and cell models. Therefore, this study demonstrates the potential promotional effect of TRIM35 in the renal fibrosis process, which may prove to be a new biomarker for the diagnosis and development of new treatments of renal fibrosis.
Open Access
Diallyl trisulfide regulates cell apoptosis and invasion in human osteosarcoma U2OS cells through regulating PI3K/AKT/GSK3β signaling pathway
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) He, Pan; Wang, Zhijun; Sheng, Bin; Xu, Yongqiang; Feng, Siyin; Huang, Yan; Gong, Fuqiang; Tang, Liting; Xie, Liming
Aims. To investigate the effects and the mechanisms of action of Diallyl trisulfide (DATS) on the proliferation and metastasis of human osteosarcoma (OS) U2OS. Methods. U2OS cells were treated by different concentrations of DATS at different time points. Cell proliferations were measured by MTT assay. DATS induced cell cycle distribution and apoptosis were evaluated by flow cytometry (FCM) with Annexin-V. Cell migration and invasion were detected by wound healing assay and transwell assay. The effects of DATS in U2OS cell growth and metastasis were also detected in a mouse OS xenograft model. Results. A time- and concentration-dependent cytotoxic effect of DATS was observed in U2OS cells. FCM with PI staining and Annexin-V -FITC indicated that DATS induces apoptosis and a G0/G1 cell cycle arrest of U2OS cells at all concentrations from 25 μmol/l to 100 μmol/l. DATS also inhibits the migration and invasion of U2OS cells. Western blot showed that the expression levels of p-AKT, p-GSK3β, Bcl-2, Vimentin and β-catenin were decreased, while the expression levels of Bad, Bax and E-cadherin were significantly increased in DATS treated U2OS cells. Analysis using a mouse xenograft model indicated that xenografts of DATS treatment group had a significant decrease in tumor volume and weight compared to the control group. Lung metastasis models in mice demonstrated that treatment of DATS inhibits lung metastasis of OS in vivo. Conclusions. These data suggested that DATS inhibits OS development and progression through the regulation of PI3K/AKT/GSK3β signaling pathways, accompanied by downregulation of Bcl-2, Vimentin and β-catenin, as well as upregulation of Bad, Bax and Ecadherin. Therefore, our data demonstrated that DATS exerted its anticancer effects by inhibiting cell proliferation, migration and invasion in vitro and in vivo. These results provide evidence for the use of the natural product DATS either alone or in combination with standard therapy for OS.
Open Access
Down-regulation of miR-215 attenuates lipopolysaccharide-induced inflammatory injury in CCD-18co cells by targeting GDF11 through the TLR4/NF-κB and JNK/p38 signaling pathways
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Sun, Boyang; Xing, Kai; Qi, Chen; Yan, Ke; Xu, Yan
Ulcerative colitis (UC) is a risk factor for carcinogenesis of colorectal cancer, which is associated with disruption of the epithelial barrier and disorder of the inflammatory response. It has been reported that the expression of microRNA (miR)-215 is upregulated in patients with long-term UC. The present study aimed to investigate the effects of miR-215 on lipopolysaccharide (LPS)-induced inflammatory injury in CCD-18Co cells, as well as to identify the underlying possible molecular mechanisms. CCD-18Co cells were treated with 1 µg/ml LPS to induce inflammatory injury. Reverse transcription-quantitative PCR was performed to determine the expression of miR-215 in LPS-treated CCD-18Co cells. Moreover, a dual luciferase reporter system assay was used to evaluate the interaction of miR-215 and growth differentiation factor 11 (GDF11) in CCD-18Co cells. The expression of miR-215 was significantly upregulated in LPS-treated CCD-18Co cells. Knockdown of miR-215 significantly alleviated the inflammatory response and oxidative stress in LPStreated CCD-18Co cells. In addition, GDF11 was identified as a direct binding target of miR-215 in CCD18Co cells. Knockdown of miR-215 significantly increased the expression of GDF11, but decreased the expression levels of Toll-like receptor (TLR)4, phosphorylated (p)-p65, iNOS, p-p38 and p-JNK in LPS-treated CCD-18Co cells. Collectively, the present findings indicated that knockdown of miR-215 alleviated oxidative stress and inflammatory response in LPStreated CCD-18Co cells by upregulating GDF11 expression and inactivating the TLR4/NF-κB and JNK/p38 signaling pathways.
Open Access
Expressions of IL-8 and CXCL5 in uterine endometrioid carcinomas which have frequent neutrophil infiltration and comparison to colorectal adenocarcinoma
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Mochizuki, Kunio; Oishi, Naoki; Kawai, Masataka; Odate, Toru; Tahara, Ippei; Inoue, Tomohiro; Kasai, Kazunari; Kondo, Tetsuo
In endometrioid carcinomas (ECs) of the uterine corpus, neutrophil accumulation within the carcinoma cell clusters is a representative microscopic finding. Because this accumulation is active, some sort of transmitter ought to exist between the EC cells and neutrophils. Interleukin-8 (IL-8) and C-X-C motif chemokine ligand 5 (CXCL5) is a cytokine that attracts neutrophils in vivo. In this study, we investigated IL-8, CXCL5 and C-X-C motif chemokine receptor 2 (CXCR2) (their chemokine receptor) expressions in ECs by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). There are few reports on the relationship between these chemokines and ECs. For comparison, we enrolled samples of colorectal adenocarcinoma (CRAC), it is another representative tumor with neutrophil infiltration. We analyzed 30 ECs and 30 CRACs. We confirmed IL-8 expression (H-score ≥50 points) in 40% of EC and 7% of CRAC samples; CXCL5 expression in 7% of EC and 10% of CRAC samples; CXCR2 expression in 83% of EC and 53% of CRAC samples by immunohistochemistry. We examined each mRNA (IL-8 and CXCL5) expression of 3 representative EC and 3 CRAC samples. Finding IL-8 expression might indicate that this cytokine is important for the process of neutrophil accumulation, particularly within ECs. The participation of CXCL5 regarding neutrophil accumulation within their carcinoma cell clusters might be restrictive compared to IL-8.
Open Access
Transplantation of mesenchymal stem cells preserves podocyte homeostasis through modulation of parietal epithelial cell activation in adriamycin-induced mouse kidney injury model
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Aslam, Rukhsana; Hussain, Ali; Cheng, Kang; Kumar, Vinod; Malhotra, Ashwani; Gupta, Sanjeev; Singhal, Pravin C.
To determine the role of the transplantation of bone marrow-derived mesenchymal stem cells (MSCs) in podocyte renewal, we studied BALB/C mice with or without adriamycin-induced acute kidney injury. MSCs were transplanted ectopically under the capsule of the left kidney or into the peritoneal cavity after the onset of kidney injury to test their local or systemic paracrine effects, respectively. Adriamycin produced increases in urine protein: creatinine ratios, blood urea nitrogen, and blood pressure, which improved after both renal subcapsular and intraperitoneal MSCs transplants. The histological changes of adriamycin kidney changes regressed in both kidneys and in only the ipsilateral kidney after intraperitoneal or renal subcapsular transplants indicating that the benefits of transplanted MSCs were related to the extent of paracrine factor distribution. Analysis of kidney tissues for p57-positive podocytes showed that MSC transplants restored adriamycin-induced decreases in the abundance of these cells to normal levels, although after renal subcapsular transplants these changes did not extend to contralateral kidneys. Moreover, adriamycin caused inflammatory activation of PECs with coexpression of CD44 and phospho-ERK, which was normalized in both or only ipsilateral kidneys depending on whether MSCs were transplanted in the peritoneal cavity or subcapsular space, respectively
Open Access
Absence of lymphatic vessels in non-functioning bleb capsules of glaucoma drainage devices
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Siggel, Robert; Schroedl, Falk; Dietlein, Thomas; Koch, Konrad R.; Platzl, Christian; Kaser-Eichberger, Alexandra; Cursiefen, Claus; Heindl, Ludwig M.
Purpose. To evaluate the presence and appearance of blood and lymphatic vessels in nonfunctioning bleb capsules of glaucoma drainage devices (GDD). Materials and methods. Non-functioning (n=14) GDD-bleb capsules of 12 patients were analyzed by immunohistochemistry for blood vessels (CD31, vascular endothelium), lymphatic vessels (lymphatic vessel endothelial hyaluronan receptor-1 [LYVE-1] and podoplanin) and macrophages (CD68). Results. CD31+++ blood vessels and CD68+ macrophages were detected in the outer layer of all specimens. LYVE-1 immunoreactivity was registered in single non-endothelial cells in 8 out of 14 (57%) bleb capsule specimens. Podoplanin-immunoreactivity was detected in all cases, located in cells and profiles of the collagen tissue network of the outer and/or the inner capsule layer. However, a colocalization of LYVE-1 and podoplanin as evidence for lymphatic vessels was not detected. Conclusions. We demonstrate the presence of bloodvessels but absence of lymphatic vessels in nonfunctioning bleb capsules after GDD-implantation. While the absence of lymphatic vessels might indicate a possible reason for drainage device failure, this needs to be confirmed in upcoming studies, including animal experiments.

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