== Panel A: histology images of iNOS expression (red fluorescence), DAPI nuclear staining (blue fluorescence), and merged images from the immunofluorescence of iNOS and DAPI. chain kinase (MLCK), and MLCK mRNA expression in jejunal segments of CP rats and down-regulated those increased parameters in DP rats. Taken with each other, atractylodin alleviated rat jejunal inflammation and exerted contractile-state-dependent regulation around the contractility of jejunal segments isolated from CP and DP rats respectively, suggesting the potential clinical implication to get ameliorating intestinal inflammation and co-occurring AZD7986 dysmotility. Keywords: Atractylodin, Constipation, Contractile-state-dependent regulation, Diarrhea, Inflammation, Jejunal contractility == INTRODUCTION == Atractylodis Rhizoma (Cangzhu) is widely used as a traditional herbal medicine [1, 2]. Cangzhu contains essential oils, phenolic acids, sesquiterpenes, and polyethylene alkynes [3, 4, 5]. Atractylodin, one of the polyethylene alkynes with Hepatoprotective, anti-oxidative, and anti-obesity effects, is used to ameliorate rheumatic diseases and night blindness [6, 7]. Atractylodin is also used to improve digestive disorders and the delay of L-NNA-induced gastric emptying [8]. Inflammatory bowel disease (IBD) and irritable bowel syndrome (IBS) are major intestinal disorders and both IBD and IBS are characterized by having inflammation and the co-occurring intestinal dysmotility in different extent [9, 10]. Diarrhea, constipation and intestinal inflammation are the main symptoms of the patients with IBD and IBS [11, 12]. Clinical and epidemiological studies indicate that the inflammation is found from the jejunum to the rectum in both IBS and IBD [13, 14, 15, 16, 17]. Diarrhea is the common symptoms of IBD; IBS may possess constipation-predominant symptom or diarrhea-predominant symptom [18, 19, 20]. The differential diagnosis for distinguishing IBD and IBS based on clinical signs and symptoms are hard [21, 22, 23]. The above observations suggest that furtherance of intestinal inflammation and co-occurring dysmotility are required to get the treatment of IBD and IBS. However , simultaneous improvement of intestinal inflammation and the co-occurring dysmotility using current available drug in clinic is rarely reported. Our pre-experiments indicated that atractylodin ameliorated intestinal inflammation and exerted beneficial modulation on intestinal contractility in rats. The present study was designed to characterize the effects of atractylodin on intestinal inflammation and the co-occurring intestinal dysmotility and to uncover the mechanisms by using both diarrhea prominent (DP) rat and constipation prominent (CP) rat versions. == METHODS == == Animals == Forty male rats of SpragueDawley (SD) weighing 180~220 g were provided by the Experimental Creature Center of Dalian Medical University (Dalian, China). The experiment protocol was carried out based on the Declaration of Helsinki, and supported by Dalian Medical University Animal Treatment and Ethics AZD7986 Committee. Every six rats were housed in a crate and put in a temperature-controlled room with a 12-hlight-dark cycle. Food and water were available for ad libitum consumption. == Experimental models of diarrhea and constipation == DP rats were established by intracolonic instillation of 4. 0% (V/V) acetic acid and restraint stress, and the control rats underwent by intracolonic instillation with saline; CP rats were established by daily gavage with cool water (0~4) to get 14 days and the control rats gavaged with water at room heat [24, 25, 26, 27, 28]. The successful establishment of both DP and CP rats was confirmed by the determination of intestinal inflammation in both rat versions and by the observation from the increased jejunal contractility in DP rats and decreased jejunal contractility in CP rats. Intragastric gavage atractylodin (10. 0 mg/kg) to get 7 days were performed to get both DP and CP AZD7986 rats and normal rat controls were given same amount of vehicle. Jejunal segments isolated from control rats, DP rats, and CP rats were collected and used in the determination of jejunal contractility and evaluate the effects of atractylodin [29]; the jejunal segments isolated from DP rats, atractylodin-treated DP rats, CP rats, atractylodin-treated CP rats, and control rats were used in the determination of mRNA expression of myosin light chain kinase (MLCK), protein content of MLCK, and phosphorylation extent from the 20 kDa myosin light chain (p-MLC20). The granules and moisture content from Rabbit Polyclonal to ATP5I the feces from each group were calculated daily. == ELISA assay == Serum pro-inflammatory cytokines and mediators, including tumor necrosis factor-alpha (TNF-), Interleukin-1-beta AZD7986 (IL-1), and Interleukin-6 (IL-6) were identified using double-antibody sandwich ELISAs (R&D Systems, USA). == Intestinal histopathology == The segments of jejunum were detached to assess the morphological changes of the jejunum.