Sympathetic Denervation Alters the Inflammatory Response of Resident Muscularis Macrophages upon Surgical Trauma and Ameliorates Postoperative Ileus in Mice

Interactions between the peripheral nervous system and resident macrophages (MMs) modulate intestinal homeostatic functions. Activation of β2-adrenergic receptors on MMs has been shown to reduce bacterial challenges. These MMs are also crucial for the development of bowel inflammation in postoperative ileus (POI), an iatrogenic, noninfectious inflammation-based motility disorder. However, the role of the sympathetic nervous system (SNS) in the immune modulation of these MMs during POI or other noninfectious diseases is largely unknown. By employing 6-OHDA-induced denervation, we investigated the changes in the muscularis externa by RNA-seq, quantitative PCR, and flow cytometry. Further, we performed transcriptional phenotyping of sorted CX3CR1+ MMs and ex vivo LPS/M-CSF stimulation on these MMs. By combining denervation with a mouse POI model, we explored distinct changes on CX3CR1+ MMs as well as in the muscularis externa and their functional outcome during POI. Our results identify SNS as an important mediator in noninfectious postoperative inflammation. Upon denervation, MMs anti-inflammatory genes were reduced, and the muscularis externa profile is shaped toward a proinflammatory status. Further, denervation reduced MMs anti-inflammatory genes also in the early phase of POI. Finally, reduced leukocyte infiltration into the muscularis led to a quicker recovery of bowel motility in the late phase of POI.

Histological Structure of Varanus Salvator Intestine

Varanus salvator is a common species in Indonesia. This animal is known as the water lizard, who is a member of the family Varanidae or known as the monitor lizard. This research aims to find out histological structure of Varanus salvator intestine. Histological observation was carried out by making histological preparations using the paraffin method. The results of this study were analyzed in a comparative descriptive manner through observation in a microscope and transverse photo histology preparations using a camera. The results of this study indicate that the intestinal histological structure of Varanus salvator consists of Serosa (Sr), Muscle layer (Ml), Muscularis externa (Me), Submucosa (Sm), Mucosa (M), Blood Vessel (Bv) dan Lamina Propria (Lp).

Histological aspects of the esophagus in guinea pigs (Cavia porcellus)

The aim of this study is the detailed description of the esophageal microscopic structure in guinea pigs. Fragments from the cervical, thoracic and abdominal segments of the esophagus were processed for classical paraffin embedding and the sections were stained by hematoxylin-eosin and Goldner’s trichrome methods. The esophagus in guinea pigs is lined by keratinized stratified squamous epithelium and there are no glands in the lamina propria and the submucosa. The muscularis mucosae is formed by smooth muscle cells disposed longitudinally. The muscularis externa is formed by striated muscle cells disposed on 3 layers in the first 2 thirds of the esophagus, while in some regions of the abdominal segment’s circumference, the cells form 4 layers.

microRNA overexpression in slow transit constipation leads to reduced NaV1.5 current and altered smooth muscle contractility

ObjectiveThis study was designed to evaluate the roles of microRNAs (miRNAs) in slow transit constipation (STC).DesignAll human tissue samples were from the muscularis externa of the colon. Expression of 372 miRNAs was examined in a discovery cohort of four patients with STC versus three age/sex-matched controls by a quantitative PCR array. Upregulated miRNAs were examined by quantitative reverse transcription qPCR (RT-qPCR) in a validation cohort of seven patients with STC and age/sex-matched controls. The effect of a highly differentially expressed miRNA on a custom human smooth muscle cell line was examined in vitro by RT-qPCR, electrophysiology, traction force microscopy, and ex vivo by lentiviral transduction in rat muscularis externa organotypic cultures.ResultsThe expression of 13 miRNAs was increased in STC samples. Of those miRNAs, four were predicted to target SCN5A, the gene that encodes the Na+ channel NaV1.5. The expression of SCN5A mRNA was decreased in STC samples. Let-7f significantly decreased Na+ current density in vitro in human smooth muscle cells. In rat muscularis externa organotypic cultures, overexpression of let-7f resulted in reduced frequency and amplitude of contraction.ConclusionsA small group of miRNAs is upregulated in STC, and many of these miRNAs target the SCN5A-encoded Na+ channel NaV1.5. Within this set, a novel NaV1.5 regulator, let-7f, resulted in decreased NaV1.5 expression, current density and reduced motility of GI smooth muscle. These results suggest NaV1.5 and miRNAs as novel diagnostic and potential therapeutic targets in STC.

Comparative Histological Study of the Stomach in Two Species of Iraqi Vertebrates (Magpie Pica pica L. and Small Asian Mongoose Herpestes javanicus E.)

A histological study showed the wall of the stomach in Pica pica and Herpestes javanicus consists of four layers: mucosa, submucosa, muscularis externa and serosa. Also, the present study showed many differences in the histological structures of the stomach for each in both types. The stomach of P. pica consists of two portions: the proventiculus and gizzard, while the stomach of H. javanicus consists of three portions: cardiac, fundic and pyloric regions. The mucosa layer formed short gastric folds, named plicae. In the proventiculus of P. pica, sulcus is found between each two plicae, but the folds called gastric pits in the gizzard, which are full with koilin. Lamina properia in both types contained gastric glands (straight simple tubular glands) named superficial glands, as well as another gastric gland found in the submucosa layer of the proventiculus in P. pica only named deep gastric glands. The gastric gland in the stomach of H. javanicus contained: mucous neck cells and parietal cells positive to AB/PAS stains in cardiac portion, as well as chief cells in fundic portion, but pyloric portion had just mucous neck cells. Muscularis externa in both types formed two muscle layers: inner and outer layer.

Comparative Histological Study of the Stomach in Two Species of Iraqi Vertebrates (Magpie Pica pica L. and Small Asian Mongoose Herpestes javanicus E.)

A histological study showed the wall of the stomach in Pica pica and Herpestes javanicus consists of four layers: mucosa, submucosa, muscularis externa and serosa. Also, the present study showed many differences in the histological structures of the stomach for each in both types. The stomach of P. pica consists of two portions: the proventiculus and gizzard, while the stomach of H. javanicus consists of three portions: cardiac, fundic and pyloric regions. The mucosa layer formed short gastric folds, named plicae. In the proventiculus of P. pica, sulcus is found between each two plicae, but the folds called gastric pits in the gizzard, which are full with koilin. Lamina properia in both types contained gastric glands (straight simple tubular glands) named superficial glands, as well as another gastric gland found in the submucosa layer of the proventiculus in P. pica only named deep gastric glands. The gastric gland in the stomach of H. javanicus contained: mucous neck cells and parietal cells positive to AB/PAS stains in cardiac portion, as well as chief cells in fundic portion, but pyloric portion had just mucous neck cells. Muscularis externa in both types formed two muscle layers: inner and outer layer.

A comparative study about the immunomodulatory effects of tramadol and metamizole in a murine model of postoperative ileus

Postoperative ileus (POI) is a common complication after abdominal surgery characterized by motility disturbances leading to increased morbidity and mortality in surgical patients. Intestinal manipulation of the murine small bowel is an established animal model resulting in an increased postsurgical inflammation within the intestinal muscular externa and a delayed gastrointestinal transit. Some analgesics have been shown to affect inflammation. In this study, we compared the immunomodulatory effects of two different analgesics. Mice were treated with tramadol, metamizole or saline as a control in our established POI model. The postoperative inflammatory response was assessed by gene expression of pro-inflammatory cytokines at different time points and immunocytes extravasation into the muscularis externa. Functional motility analyses were performed by a gastrointestinal transit measurement. Metamizole application reduced the pro-inflammatory response after surgery and improved gastrointestinal motility, while tramadol showed no alteration in cytokine gene expression, influx of immunocytes and gastrointestinal transit compared with the controls. In conclusion. we suggest tramadol as analgesia in immunological studies on POI in mice as it does not affect the underlying inflammation of POI.

Immunohistochemical localization of adropin in the small intestine of rats

The aim of this study was to determine the occurrence and distribution of adropin in the small intestine in control and gastrectomized rats. The study was carried out on 12 Wistar rats. Six control rats (SHO) underwent a sham operation. Six rats were subjected to gastrectomy (Gx); i.e. resection of the antrum and the glandular part of the stomach. Six weeks after the surgery, the rats were sacrificed. The duodenum and middle part of the jejunum were collected for immunohistochemical and immunofluorescence procedures. An immunopositive reaction to ADR was detected in the duodenum and jejunum, in crypts, and in enterocytes located along the entire length of the villi. Furthermore, more intense ADR immunoreactivity was observed in the crypts and villi of the duodenum than in the jejunum. The presence of adropin was also detected in the submucosa and muscularis externa of the duodenum and jejunum, in the cells of Brunner’s gland of the duodenum, and in myenteric and submucosal plexus nerve. The ADR immunoreactivity in the crypts and villi in the duodenum and jejunum was lower in the Gx rats compared to the SHO rats. The ADR distribution in the duodenum and jejunum has also been confirmed in immunofluorescence studies. In conclusion, our data demonstrated ADR expression in all histological layers of the small intestine in the rats. Gx reduced ADR immunoreactivity in the crypts and enterocytes. This may suggest a potential impact of ADR on intestine function such as absorbability and immune reactions. However, further research is needed to explain the ADR role in gastrointestinal functions