Changes in apoptotic factors in hypothalamus and hippocampus after acute and subchronic stress induction during conditioned place preference paradigm

The hypothalamus (HYP) and hippocampus (HIP) are important regions involved in stress responses. These areas are also associated with reward processing. In this study, the effects of acute and subchronic stress on the changes in apoptotic factors (Bax/Bcl-2 ratio, caspase-3 activation and PARP degradation) in the HYP and HIP during conditioned place preference (CPP) paradigm were evaluated. Male Wistar rats were divided into two saline- and morphine-reated supergroups. Each supergroup contained control, acute stress (AS) and subchronic stress (SS) groups. In all groups, CPP paradigm was done and thereinafter alterations of apoptotic factors were measured by western blot. The results revealed that in the HYP, all mentioned factors increased significantly in saline- or morphine-treated animals during AS and SS. On the other hand, in the HIP, Bax/Bcl-2 ratio in saline-treated animals increased significantly during AS and SS, while in morphine-treated animals this ratio did not have any significant alteration during AS and was decreased during SS compared with morphine-control group. Caspase-3 and PARP increased during AS and SS in saline- or morphine-treated animals. For example, caspase-3 increased during AS and SS in morphine-treated animals by 2.4 folds and PARP (89 KDa) increased by 3.1 and 3.5 folds, respectively. Interestingly, the increase of apoptotic factors in morphine-treated animals was more considerable than that of saline-treated animals. These results strongly implied that AS and SS trigger apoptotic events in the HYP and HIP of saline- and/or morphine-treated animals as well as morphine reinforces the effect of stress on the induction of apoptosis

The anti-apoptotic and anti-inflammatory effect of Lactobacillus acidophilus on Shigella sonnei and Vibrio cholerae interaction with intestinal epithelial cells: A comparison between invasive and non-invasive bacteria.

The aim of this study was to compare the effect of Lactobacillus acidophilus on the attachment, invasion, and interaction of Shigella sonnei and Vibrio cholerae with Caco-2 epithelial cells. Also, the anti-apoptotic and anti-inflammatory effect of L. acidophilus was investigated on S. sonnei and V. cholerae interaction with Caco-2 cells as the representatives of invasive and non-invasive intestinal bacteria. It was found that pretreatment with L. acidophilus significantly prevented from adherence and internalization of S. sonnei/V. cholerae and reduced the expression of tumour necrosis factor-α and interleukin-8 in host cells. No significant difference was observed in inhibitory effect of Lactobacilli in V. cholerae and S. sonnei attachment, emphasizing on the role of lactobacilli as a physical barrier in inhibiting direct contact with host cell by competitive exclusion, which may affect attachment and subsequent internalization of both invasive and non-invasive pathogenic bacteria in a same scale. The evaluation of early and late apoptosis in Caco-2 cells exposed to V. cholerae/S. sonnei and pretreated by L. acidophilus indicated no remarkable difference in L. acidophilus anti-apoptotic effect on Caco-2 cells against invasive and non-invasive bacterial infection. Moreover, L. acidophilus by itself showed no apoptotic effect on Caco-2 cells. Statistical analysis revealed that L. acidophilus in S. sonnei infected cells was able to reduce pro-inflammatory immune responses (TNF-α, IL-8 and IL-1β) and NO and PGE2 secretion more strongly compared with V. cholerae infected cells. These data showed for the first time that the protective effect of Lactobacilli, as a probiotic bacterium, in interaction suppression was more in invasive bacteria including S. sonnei than in non-invasive V. cholerae

Association between Probiotics and Modulation of Gut Microbial Community Composition in Colorectal Cancer Animal Models: A Systematic Review (2010–2021)

Background. Colorectal cancer (CRC) is one of the most prevalent gastrointestinal malignancies and is considered the third major cause of mortality globally. Probiotics have been shown to protect against the CRC cascade in numerous studies. Aims. The goal of this systematic review was to gather the preclinical studies that examined the impact of probiotics on the alteration of gut microbiota profiles (bacterial communities) and their link to colorectal carcinogenesis as well as the potential processes involved. Methods. The search was performed using Scopus, Web of Science, and PubMed databases. Five parameters were used to develop search filters: “probiotics,” “prebiotics,” “synbiotics,” “colorectal cancer,” and “animal model.” Results. Of the 399 full texts that were screened, 33 original articles met the inclusion criteria. According to the current findings, probiotics/synbiotics could significantly attenuate aberrant crypt foci (ACF) formation, restore beneficial bacteria in the microbiota population, increase short-chain fatty acids (SCFAs), and change inflammatory marker expression. Conclusions. The present systematic review results indicate that probiotics could modulate the gut microbial composition and immune regulation to combat/inhibit CRC in preclinical models. However, where the evidence is more limited, it is critical to transfer preclinical research into clinical data

The anti-apoptotic and anti-inflammatory effect of <i>Lactobacillus acidophilus</i> on <i>Shigella sonnei</i> and <i>Vibrio cholerae</i> interaction with intestinal epithelial cells: A comparison between invasive and non-invasive bacteria

<div><p>The aim of this study was to compare the effect of <i>Lactobacillus acidophilus</i> on the attachment, invasion, and interaction of <i>Shigella sonnei</i> and <i>Vibrio cholerae</i> with Caco-2 epithelial cells. Also, the anti-apoptotic and anti-inflammatory effect of <i>L</i>. <i>acidophilus</i> was investigated on <i>S</i>. <i>sonnei</i> and <i>V</i>. <i>cholerae</i> interaction with Caco-2 cells as the representatives of invasive and non-invasive intestinal bacteria. It was found that pretreatment with <i>L</i>. <i>acidophilus</i> significantly prevented from adherence and internalization of <i>S</i>. <i>sonnei</i>/<i>V</i>. <i>cholerae</i> and reduced the expression of tumour necrosis factor-α and interleukin-8 in host cells. No significant difference was observed in inhibitory effect of Lactobacilli in <i>V</i>. <i>cholerae</i> and <i>S</i>. <i>sonnei</i> attachment, emphasizing on the role of lactobacilli as a physical barrier in inhibiting direct contact with host cell by competitive exclusion, which may affect attachment and subsequent internalization of both invasive and non-invasive pathogenic bacteria in a same scale. The evaluation of early and late apoptosis in Caco-2 cells exposed to <i>V</i>. <i>cholerae/S</i>. <i>sonnei</i> and pretreated by <i>L</i>. <i>acidophilus</i> indicated no remarkable difference in <i>L</i>. <i>acidophilus</i> anti-apoptotic effect on Caco-2 cells against invasive and non-invasive bacterial infection. Moreover, <i>L</i>. <i>acidophilus</i> by itself showed no apoptotic effect on Caco-2 cells. Statistical analysis revealed that <i>L</i>. <i>acidophilus</i> in <i>S</i>. <i>sonnei</i> infected cells was able to reduce pro-inflammatory immune responses (TNF-α, IL-8 and IL-1β) and NO and PGE₂ secretion more strongly compared with <i>V</i>. <i>cholerae</i> infected cells. These data showed for the first time that the protective effect of Lactobacilli, as a probiotic bacterium, in interaction suppression was more in invasive bacteria including <i>S</i>. <i>sonnei</i> than in non-invasive <i>V</i>. <i>cholerae</i>.</p></div

Annexin V-FITC/PI staining and flow cytometry assay to determine the proportion of apoptosis after pretreatment of Caco-2 cells with <i>L</i>. <i>acidophilus</i> and then infected by <i>V</i>. <i>colarae/ S</i>. <i>sonnei</i>.

<p>Images showed the typical morphologies of cells in the different regions. (A) Q1: Necrotic cells, Q2: Late apoptotic cells, Q2: Early apoptotic cells and Q4: Viable cells. <i>L</i>.<i>a</i>: <i>Lactobacillus acidophilus</i>, <i>V</i>.<i>c</i>: <i>Vibrio cholerae</i> and <i>S</i>.<i>s</i>: <i>Shigella sonnei</i>. (B)The pretreatment with <i>L</i>. <i>acidophilus</i> on Caco-2 cells exposed to <i>V</i>. <i>colarae/ S</i>. <i>sonnei</i> resulted in decreased apoptosis. Statistical analysis of representative images was performed with three independent experiments. The statistical significances were achieved when P <0.05 (***P <0.001). *** significantly different from control cells. ^### significantly different of <i>L</i>.<i>a</i>+<i>V</i>.<i>c</i>+Caco-2 from <i>V</i>.<i>c</i>+Caco-2 cells. ^††† significantly different of <i>L</i>.<i>a</i>+<i>S</i>.<i>s</i>+Caco-2 from <i>S</i>.<i>s</i>+Caco-2 cells <i>L</i>.<i>a</i>: <i>Lactobacillus acidophilus</i>, <i>V</i>.<i>c</i>: <i>Vibrio cholerae</i> and <i>S</i>.<i>s</i>: <i>Shigella sonnei</i>. ns: none significant.</p

The effect of <i>V</i>. <i>cholerae</i>, <i>S</i>. <i>sonnei</i> and <i>L</i>. <i>acidophilus</i> on cell viability.

<p>(A) cell viability of Caco-2 cells infected by <i>V</i>. <i>colarae</i>/ <i>S</i>. <i>sonnei</i> was determined by MTT assay, Each value represents the mean ±SEM (n = 3). ***p < 0.001 compared with control (untreated Caco-2cells) by one way ANOVA with Tukey’s post hoc test. ^###p < 0.001 significantly different of <i>L</i>.<i>a</i>+<i>V</i>.<i>c</i>+Caco-2 group from <i>V</i>.<i>c</i>+Caco-2 group. ^†††p < 0.001 significantly different between <i>L</i>.<i>a</i>+<i>S</i>.<i>s</i>+Caco-2 group with <i>S</i>.<i>s</i>+Caco-2 (by unpaired two-tailed Student’s <i>t</i> test). All experiments were repeated three times. <i>L</i>.a: <i>Lactobacillus acidophilus</i>, <i>V</i>.<i>c</i>: <i>Vibrio cholerae</i> and <i>S</i>.<i>s</i>: <i>Shigella sonnei</i>, ns: none significant. (B) Morphological evaluation of Caco-2 cells under the above-mentioned treatment by light microscopic observation. <i>L</i>.a: <i>Lactobacillus acidophilus</i>, <i>V</i>.<i>c</i>: <i>Vibrio cholerae</i> and <i>S</i>.<i>s</i>: <i>Shigella sonnei</i>.</p

The anti-apoptotic and anti-inflammatory effect of <i>Lactobacillus acidophilus</i> on <i>Shigella sonnei</i> and <i>Vibrio cholerae</i> interaction with intestinal epithelial cells: A comparison between invasive and non-invasive bacteria - Fig 2

<p>(<b>A and B) Effect of pretreatment of <i>L</i>. <i>acidophilus</i> on its ability to inhibit <i>V</i>. <i>colarae/ S</i>. <i>sonnei</i> adherence and invasion to Caco-2 cells.</b> ND means Not Detectable. Each value indicates the mean±SEM from three independent experiments and three replicates (n = 3). ^###p < 0.001 significantly different of <i>L</i>.<i>a</i>+<i>V</i>.<i>c</i>+Caco-2 group from <i>V</i>.<i>c</i>+Caco-2 group. ^†††p < 0.001 significant difference between <i>L</i>.<i>a</i>+<i>S</i>.<i>s</i>+Caco-2 group and <i>S</i>.<i>s</i>+Caco-2 (by unpaired two-tailed Student’s <i>t</i> test). <i>L</i>.<i>a</i>: <i>Lactobacillus acidophilus</i>, <i>V</i>.<i>c</i>: <i>Vibrio cholerae</i> and <i>S</i>.<i>s</i>: <i>Shigella sonnei</i>.</p

NO level in Caco-2 cells after <i>L</i>. <i>acidophilus</i> pretreatment and followed by <i>V</i>. <i>cholerae/ S</i>. <i>sonnei</i>.

<p>NO production after 24h, significantly decreased in comparison with <i>V</i>.<i>cholerae/ S</i>. <i>sonnei</i> infected cells alone. The mean of three independent experiments is shown. The significant differences were notabled when P <0.05. (***P <0.001) significantly different from control cells. ^##significantly different of <i>L</i>.<i>a</i>+<i>V</i>.<i>c</i>+Caco-2 from <i>V</i>.<i>c</i>+Caco-2 cells. ^††† significantly different of <i>L</i>.<i>a</i>+<i>S</i>.<i>s</i>+Caco-2 from <i>S</i>.<i>s</i>+Caco-2 cells <i>L</i>.<i>a</i>: <i>Lactobacillus acidophilus</i>, <i>V</i>.<i>c</i>: <i>Vibrio cholerae</i> and <i>S</i>.<i>s</i>: <i>Shigella sonnei</i>. ns: none significant.</p

Effect of treatment with <i>L</i>. <i>acidophilus</i> on PGE2 secretion in Caco-2 epithelial cells.

<p>PGE2 secretion was significantly elevated following infection with <i>V</i>. <i>cholerae/ S</i>. <i>sonnei</i> and this effect was abolished when the cells were pretreated by <i>L</i>. <i>acidophilus</i>. The data are presented as mean ±SEM. The statistical data (by unpaired two-tailed Student’s <i>t</i> test) were signed when P <0.05. ^##significantly different of <i>L</i>.<i>a</i>+<i>V</i>.<i>c</i>+Caco-2 from <i>V</i>.<i>c</i>+Caco-2 cells. ^††† significantly different of <i>L</i>.<i>a</i>+<i>S</i>.<i>s</i>+Caco-2 from <i>S</i>.<i>s</i>+Caco-2 cells <i>L</i>.<i>a</i>: <i>Lactobacillus acidophilus</i>, <i>V</i>.<i>c</i>: <i>Vibrio cholerae</i> and <i>S</i>.<i>s</i>: <i>Shigella sonnei</i>. ND: None Detectable.</p

Changes in apoptotic factors caspase-3, PARP and Bax/Bcl-2 ratio in the ventral tegmental area after the acquisition and extinction of morphine-induced conditioned place preference in the rat

Introduction: Chronic high doses of morphine inhibit cell proliferation and induce cell death. One of the centers in reward pathway is the ventral tegmental area (VTA). Stimulation of opioid receptors in the reward circuit in the brain by morphine could be enabling a factor induce apoptosis in some brain regions, and expression of death receptors increases on the cell surface. This study was designed to evaluate the effect of morphine on changes in apoptotic factors (Bax/Bcl-2, PARP and caspase-3) in the VTA during the acquisition of morphine-induced conditioned place preference (CPP) and extinction period. Materials and Methods: In this study, in behavioral experiments, the CPP paradigm was done on 64 adult male albino Wistar rats. In saline-control and three doses of morphine (0.5, 5, 10 mg/kg) experimental groups in acquisition, in addition with effective dose of morphine (5 mg/kg) evaluate extinction period (8 days). Then, in the molecular section the changes in apoptotic proteins assess with western blot technique.Results: We found that apoptotic factors increase in all three experimental groups in response to morphine. However, the most response was significantly occurred at the dose of 5 mg/kg morphine. Additionally, there is no change has been seen in the apoptotic factors during the extinction period. Conclusion: It seems that morphine in all doses cause apoptosis, but quite contrary, by increasing the dose of morphine, the number of receptors involved in apoptosis increases and morphine’s neuroprotective effects are appeared