scholarly journals Expression of Autophagy-Related Factors LC3A and Beclin 1 and Apoptosis-Related Factors Bcl-2 and BAX in Osteoblasts Treated With Sodium Fluoride

2021 ◽  
Vol 12 ◽  
Author(s):  
Lin Xu ◽  
Chaonan Deng ◽  
Ying Zhang ◽  
Lina Zhao ◽  
Yan Linghu ◽  
...  

ObjectiveThis study aims to analyze the expressions of autophagy-related factors light chain 3 alpha (LC3A) and Beclin 1 and apoptosis-related factors B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (BAX) in primary osteoblasts treated with sodium fluoride (NaF).MethodsOsteoblasts were extracted from Sprague-Dawley rats and treated with 0, 2.5, 5, and 10 mg/L NaF solutions, followed by 10 mmol/L 3-methyladenine (3-MA) for 24 h. The apoptotic rate was determined by flow cytometry, and the expressions of the autophagy- and apoptosis-related factors were measured by western blotting and real-time quantitative polymerase chain reaction.ResultsThe mRNA expressions of LC3A, Beclin 1, and BAX in the NaF-treated osteoblast group were higher than those in the control group, while the protein expressions of these factors in the NaF-treated group were significantly higher than those in the control group. However, the Bcl-2 protein expression in the NaF-treated osteoblasts was significantly decreased compared to that in the control cells. After the 3-MA treatment, the protein expressions of LC3A, Beclin 1, and Bcl-2 were significantly decreased compared with those of the NaF-treated group, whereas the expression of BAX increased. Moreover, the apoptosis rate was increased after the addition of the 3-MA inhibitor.ConclusionNaF stimulation promoted autophagy and apoptosis of the osteoblasts, suggesting the involvement of fluoride damage in these processes.

2020 ◽  
Vol 14 (03) ◽  
pp. 124-127
Author(s):  
Somia Iqbal ◽  
Noman Sadiq ◽  
Saad Siddiqui ◽  
Hira Iqbal

Background: Obesity is a prevailing metabolic disorder that affects the functioning of the male reproductive system. Excessive adipose tissue enhances reactive oxygen species generation and is linked with male infertility. Spinach has demonstrated antioxidant effects. The present study was conducted to determine the antioxidant effects of spinach on sperm parameters in obese Sprague Dawley rats. Subjects and methods: This randomized control study was conducted at the animal house of the National Institute of Health Islamabad, Islamic International Medical College, Cosmesurge International Hospital, Rawalpindi, and Apollo lab, Islamabad, Pakistan from April 2016 to March 2017. Forty male Sprague Dawley rats having an age of 8 weeks and weight 160-200g were tagged from number 1 to 40. Every third rat was randomly allocated to control Group A (n=13) and remaining into the Experimental group (n=27). Rats of control Group A was given a standard diet while a high-fat diet was given to Experimental group rats to induce obesity for the duration of six weeks. Weight (g) was measured weekly and obesity was confirmed when rats attain more than 20% weight when compared with that of rats of control Group A. Then, after obesity induction, the experimental group was alienated into the obesity control group (Group B) and spinach treated group (Group C). For sample, rats of Group A and Group B were sacrificed, and the cauda epididymis of each rat was placed in a Petri dish containing normal saline and cut into pieces to allow the release of sperm and then sperm parameters (sperms concentration, motility, and morphology) were recorded under the microscope. Then, spinach (5% hot water extract) along with the persistence of fat diet was administered to Group C for 4 weeks and finally, sperm parameters were measured in this group. Results: Sperm concentration/ml, motility (%), and normal morphology (%) of Group B rats were significantly decreased as compared to Group A rats. However, sperm concentration/ml, motility (%), and normal morphology (%) of Group C (spinach treated group) rats was significantly increased (p<0.001) as compared to Group B (obesity control group) rats after administering spinach. Conclusion: The addition of Spinach in a normal diet regimen restores normal sperm morphology, improves sperm motility and concentration.


2021 ◽  
Vol 18 (6) ◽  
pp. 1219-1225
Author(s):  
Farah Dayana Rosli ◽  
Noor Hashida Hashim ◽  
Yusmin Mohd Yusuf ◽  
Khairul Osman ◽  
Siti Fatimah Ibrahim ◽  
...  

Purpose: To assess the effects of thymoquinone (TQ) on the integrity of sperm DNA in nicotineinduced sperm impairment in rats. Methods: Adult male Sprague Dawley rats were randomized into four equal groups: control group received normal saline orally for 60 days; nicotine group was subcutaneously injected with 5 mg/kg/day nicotine for 30 days and then given normal saline for the next 30 days; TQ group was given normal saline for 30 days followed by TQ at 5 mg/kg/day for 30 days; and nicotine-TQ group received 5 mg/kg of nicotine for 30 days and 5mg/kg of TQ for another 30 days. Sperm DNA breakages were evaluated using Comet assay. The expression levels of protamine 1 (PT1) and transition nuclear protein 2 (Tnp2) genes which are essential for the proper compaction of the sperm DNA were analyzed by quantitative polymerase chain reaction (qPCR). Results: Thymoquinone significantly decreased DNA fragmentation in the sperm of nicotine-treated rats. However, there was no change in PT1 gene expression. Tnp2 was downregulated in the nicotine group and slightly upregulated in nicotine-TQ group (p < 0.05). Conclusion: The results demonstrate the potential benefits of TQ in improving sperm DNA quality of nicotine-induced male infertility.


2021 ◽  
Vol 50 (3) ◽  
pp. 711-721
Author(s):  
SITI FATIMAH ROQIAH YAHAYA ◽  
NIZA SAMSUDDIN ◽  
SUHANA MAMAT ◽  
ROZITA HOD ◽  
NOR ZAMZILA ABDULLAH ◽  
...  

Nypa fruticans Wurmb. (nipa palm) sap, locally known as air nira or tuak, is a sweet natural beverage in Malaysia with antioxidant potency beneficial for human health. However, nypa sap lacks scientific attention with no toxicity study has been established. Therefore, this study was performed to evaluate the acute toxicity of nypa sap in the female Sprague Dawley rats. Twenty-five female rats were randomly divided into one control group and four treated groups. Treated groups were orally administered with doses of 5, 50, 300, and 2000 mg/kg of nypa sap, while the control group had received normal saline solution. The animals’ vital signs and mortality were recorded daily at an interval of 30 min and continued up to 14 days. Their blood samples and organs were harvested for toxicity analysis to assess liver and kidney function, lipid profile, and full blood count. The acute toxicity test via measurement of the biochemical and haematological parameters had shown that there was no significant difference between the treated and control groups. However, the blood glucose level in the treated groups (at higher doses of 300 and 2000 mg/kg, respectively) was significantly decreased. A similar trend was recorded for mean corpuscular volume (MCV) albeit in the treated group of 50 mg/kg doses. Histopathological examination of the organs showed no signs of abnormality in both treated and untreated groups. Overall, the results showed that consumption of nypa sap is potentially safe with no acute toxic effect on the laboratory rat models.


PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11577
Author(s):  
Youmin Mei ◽  
Xiang Shen ◽  
Xiaoqian Wang ◽  
Min Zhang ◽  
Qiao Li ◽  
...  

Objective This study aimed to investigate the expression of autophagy-related factors microtubule-associated protein l light chain 3 (LC3) and the apoptosis-related factors BCL2-associated X protein (Bax) and B cell lymphoma-2 (Bcl-2) in the periodontal tissue of experimental diabetic rats. These data were used to explore the potential mechanism in diabetes-induced periodontal tissue lesions. Methods A total of 32 Sprague Dawley (SD) rats were randomly assigned into diabetes (group D, n = 16) and control groups (group N, n = 16). The diabetic group was induced by intraperitoneal injection of 1% streptozotocin (STZ, 60 mg/kg) and the control group was injected with citrate buffer (0.1mol/L). Rats were sacrificed after 4 and 8 weeks of feeding and collected as D1, N1 groups and D2, N2 groups, and the maxilla were retained for analysis. The changes in periodontal tissue structure were observed by hematoxylin-eosin (HE) staining. The expression and distribution of LC3, Bax and Bcl-2 in the periodontium of the rats was detected by immunohistochemical (SP) staining. Results Diabetic rats showed several changes compared to control animals including sparse alveolar bone trabecular structure, loss of the lamina dura and absorption of the local alveolar bone. The positive expression level of LC3 in the gingival epithelial, periodontal ligament and alveolar bone of group D1 was significantly higher than in the N1, N2 and D2 groups (P < 0.05). The level of Bax expression in the group D2 rats was significantly higher than those in the N1, N2 and D1 groups (P < 0.05), while the positive degree of Bcl-2 was significantly lower than those of other groups (P < 0.001). LC3 was negatively correlated with Bax and was irrelevant with Bcl-2; Bcl-2 was not correlated with Bax. Conclusions The expression of LC3, Bax and Bcl-2 changes in the periodontal tissue of diabetic rats may indicate that autophagy and apoptotic are involved in the process of periodontal tissue damage in diabetic rats. These changes may be one of the mechanisms of periodontal tissue lesions.


2016 ◽  
Vol 94 (9) ◽  
pp. 936-946 ◽  
Author(s):  
Abdelaziz M. Hussein ◽  
Nashwa Barakat ◽  
Amira Awadalla ◽  
Mahmoud M. Gabr ◽  
Sherry Khater ◽  
...  

The present study investigated the effects of combination of ischemic preconditioning (Ipre) and adipose-derived mesenchymal stem cells (ADMSCs) on renal ischemia–reperfusion (I–R) injury in rats. 90 male Sprague Dawley rats were divided into 5 equal groups; sham operated, control (45 min left renal ischemia), Ipre group as control group with 3 cycles of Ipre just before renal ischemia, ADMSCs-treated group (as control with ADMSCs 106 cells in 0.1 mL via penile vein 60 min before ischemia time), and Ipre + ADMSCs group as ADMCs group with 3 cycles of Ipre. Ipre and ADMSCs groups showed significant decrease in serum creatinine and blood urea nitrogen (BUN) and caspase-3 and CD45 expression in kidney and significant increase in HIF-1α, SDF-1α, CD31, and Ki67 expressions in kidney compared with the control group (p < 0.05). Moreover, the Ipre + ADMSCs group showed significant decrease in serum BUN and caspase-3 and CD45 expression in kidney with significant increase in HIF-1α, SDF-1α, CD31, and Ki67 expression in kidney compared with the Ipre and ADMCs groups (p < 0.05). We concluded that Ipre potentiates the renoprotective effect of ADMSCs against renal I/R injury probably by upregulation of HIF-1α, SDF-1α, CD31, and Ki67 and downregulation of caspase-3 and CD45.


2020 ◽  
Vol 39 (9) ◽  
pp. 1178-1189
Author(s):  
S Huan ◽  
J Jin ◽  
C-x Shi ◽  
T Li ◽  
Z Dai ◽  
...  

In this study, to investigate the effect of overexpression of miR-146a on autophagy of hippocampal neurons in rats with intracerebral hemorrhage (ICH), 72 Sprague-Dawley rats were randomly divided into the sham, ICH, miR-146a agomir, and miR-146a agomir control groups. The ICH model was constructed by injection of collagenase VII. The apoptosis of hippocampal neurons was measured by TUNEL assay. The levels of LC3 and Beclin 1 were analyzed by immunohistochemistry. Mitochondrial autophagy was examined by transmission electron microscopy. The levels of LC3A, LC3B, Beclin 1, Bax, Bcl-2, and cleaved caspase 3 were examined by Western blot. Western blot was also used to evaluate the expression of nuclear factor κB signaling pathway-related factors. To examine the effect of autophagy inhibitor (3-methyladenine (3-MA)) on miR-146a-regulated apoptotic protein expression, 30 rats were further divided into the sham, ICH, miR-146a agomir, 3-MA, and miR-146a + 3-MA groups. The levels of Bax, Bcl-2, and cleaved caspase 3 were examined by Western blot. Compared with the sham group, the nerve function scores, brain water content, the percentage of apoptotic cells, and the expression levels of LC3, Beclin 1, Bax, cleaved caspase 3, and p-P65 in the hippocampus of rats in the ICH group were all significantly increased ( p < 0.05), whereas the expression levels of miR-146a, Bcl-2, and p-IκBα were markedly decreased ( p < 0.05). Mitochondrial autophagy was also evident. Furthermore, compared with the ICH group, the results of the abovementioned tests in the miR-146a agomir group were reversed. The overexpression of miR-146a inhibited the autophagy of hippocampal neurons in rats with ICH.


Author(s):  
Kuswati Kuswati ◽  
Djoko Prakosa ◽  
Brian Wasita ◽  
Nanang Wiyono

<p>BACKGROUND<br />Stress is one of the factors that cause apoptosis in neuronal cells. Centella<br />asiatica has a neuroprotective effect that can inhibit apoptosis. This study<br />aimed to examine the effect of Centella asiatica ethanol extract on B-cell<br />lymphoma 2 (Bcl-2) protein expression in the prefrontal cortex of rats.</p><p>METHODS<br />An experimental study was conducted on 34 brain tissue samples from male<br />Sprague Dawley rats exposed to chronic restraint stress for 21 days. The<br />samples were taken from following groups: non-stress group K, negative<br />control group P1 (stress + arabic gum powder), P2 (stress + C.asiatica at<br />150 mg/kgBW), P3 (stress + C.asiatica at 300 mg/kg BW), P4 (stress +<br />C.asiatica at 600 mg/kg body weight) and positive control group P5 (stress<br />+ fluoxetine at 10 mg/kgBW). The samples were made into sections that<br />were stained immunohistochemically using Bcl-2 antibody to determine the<br />percentage of cells expressing Bcl-2. Data were analyzed using one way<br />ANOVA test followed by a post - hoc test.</p><p>RESULTS<br />There were significant differences in mean Bcl-2 expression between the<br />groups receiving Centella asiatica compared with the non-stress group and<br />stress-only group (negative control group) (p&lt;0.05). The results were<br />comparable to those of the fluoxetine treatment group.</p><p>CONCLUSION</p><p>The Centella asiatica ethanol extract was able to increase Bcl-2 expression<br />in the prefrontal cortex of Sprague Dawley rats exposed to restraint stress.<br />This study suggests that Centella asiatica may be useful in the treatment of<br />cerebral stress.</p>


2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Kamal M. Matar ◽  
Yasin I. Tayem

We aimed to investigate the effect of induced hepatic and renal failure on the pharmacokinetics of topiramate (TPM) in rats. Twenty-four Sprague-Dawley rats were used in this study. Renal or hepatic failure was induced by a single i.p. dose of 7.5 mg/kg cisplatin (n=8) or 0.5 mL/kg carbon tetrachloride (CCl4) (n=8), respectively. Three days after cisplatin dose or 24 h after CCl4dose, the rats were administered a single oral dose of 20 mg/kg TPM. The plasma samples were quantified by LC-MS/MS method. Compared to control, plasma concentration-time profile in CCl4-treated and, to a lesser extent, in cisplatin-treated rats decreased more slowly particularly in the elimination phase. TPM oral clearance (CL/F) in CCl4-treated group was significantly lower than that in control (P<0.001), whereasAUC0−∞, T1/2, and Vd/F were significantly higher in CCl4-treated rats compared to the control (P<0.01). The CL/F was not significantly different between cisplatin-treated rats and control (P>0.05). However, in cisplatin-treated rats, the T1/2 and Vd/F were significantly higher than that in the control group (P<0.01). Both conditions failed to cause a significant effect onCmaxorTmax. The present findings suggest that induced hepatic or renal failure could modify the pharmacokinetic profile of TPM in the rat.


Author(s):  
Xiangyu Liu ◽  
Xiong Xue ◽  
Junsheng Tian ◽  
Xuemei Qin ◽  
Shi Zhou ◽  
...  

The objectives of this study were to compare the antidepressant effects between endurance and resistance exercise for optimizing interventions and examine the metabolomic changes in different types of skeletal muscles in response to the exercise, using a rat model of chronic unpredictable mild stress (CUMS)-induced depression. There were 32 male Sprague-Dawley rats randomly divided into a control group (C) and 3 experimental groups: CUMS control (D), endurance exercise (E), and resistance exercise (R). Group E underwent 30 min treadmill running, and group R performed 8 rounds of ladder climbing, 5 sessions per week for 4 weeks. Body weight, sucrose preference, and open field tests were performed pre and post the intervention period for changes in depressant symptoms, and the gastrocnemius and soleus muscles were sampled after the intervention for metabolomic analysis using the 1H-NMR technique. The results showed that both types of exercise effectively improved the depression-like symptoms, and the endurance exercise appeared to have a better effect. The levels of 10 metabolites from the gastrocnemius and 13 metabolites from the soleus of group D were found to be significantly different from that of group C, and both types of exercise had a callback effect on these metabolites, indicating that a number of metabolic pathways were involved in the depression and responded to the exercise interventions.


2021 ◽  
Vol 10 (6) ◽  
pp. 1323
Author(s):  
Victor Yip ◽  
M. Violet Lee ◽  
Ola M. Saad ◽  
Shuguang Ma ◽  
S. Cyrus Khojasteh ◽  
...  

Polatuzumab vedotin (or POLIVY®), an antibody–drug conjugate (ADC) composed of a polatuzumab monoclonal antibody conjugated to monomethyl auristatin E (MMAE) via a cleavable dipeptide linker, has been approved by the United States Food and Drug Administration (FDA) for the treatment of diffuse large B-cell lymphoma (DLBCL). To support the clinical development of polatuzumab vedotin, we characterized the distribution, catabolism/metabolism, and elimination properties of polatuzumab vedotin and its unconjugated MMAE payload in Sprague Dawley rats. Several radiolabeled probes were developed to track the fate of different components of the ADC, with 125I and 111In used to label the antibody component and 3H to label the MMAE payload of the ADC. Following a single intravenous administration of the radiolabeled probes into normal or bile-duct cannulated rats, blood, various tissues, and excreta samples were collected over 7–14 days post-dose and analyzed for radioactivity and to characterize the metabolites/catabolites. The plasma radioactivity of polatuzumab vedotin showed a biphasic elimination profile similar to that of unconjugated polatuzumab but different from unconjugated radiolabeled MMAE, which had a fast clearance. The vast majority of the radiolabeled MMAE in plasma remained associated with antibodies, with a minor fraction as free MMAE and MMAE-containing catabolites. Similar to unconjugated mAb, polatuzumab vedotin showed a nonspecific distribution to multiple highly perfused organs, including the lungs, heart, liver, spleen, and kidneys, where the ADC underwent catabolism to release MMAE and other MMAE-containing catabolites. Both polatuzumab vedotin and unconjugated MMAE were mainly eliminated through the biliary fecal route (>90%) and a small fraction (<10%) was eliminated through renal excretion in the form of catabolites/metabolites, among which, MMAE was identified as the major species, along with several other minor species. These studies provided significant insight into ADC’s absorption, distribution, metabolism, and elimination (ADME) properties, which supports the clinical development of POLIVY.


Sign in / Sign up

Export Citation Format

Share Document