Current Pharmaceutical Biotechnology - Volume 22, Issue 12, 2021

Pyroptosis is a new type of pro-inflammatory programmed cell death identified over the last few years and has since attracted researchers' attention. Current research demonstrates that pyroptosis's primary process is dependent on caspase-1 protein and gasdermin D protein and is accompanied by the release of inflammatory substances. However, the specific mechanisms and physiological or pathophysiological processes involved in pyroptosis are unclear. Presently, research on the occurrence, development, and treatment of various diseases associated with pyroptosis is of significant value. Pyroptosis has been implicated in the pathogenesis of atherosclerosis. For instance, numerous studies have revealed that pyroptosis is involved in the development and progression of atherosclerosis, and it is closely associated with pyroptosis of three essential cells (vascular endothelial cells, macrophages, and vascular smooth muscle cells) in atherosclerosis. The advancement of research on pyroptosis is anticipated to give insights into its application as a potential method for treating diseases. This article mainly reviews the relationship between pyroptosis and associated mechanisms factors that cause pyroptosis and the relationship between pyroptosis and various diseases, especially atherosclerosis.

Clinically, probiotics have a significant role in nutrition, immune function, and maintenance of gut homeostasis. Unfortunately, the widespread use of antibiotics disrupts the symbiotic balance of gut microbiota, leading to the development of several gastrointestinal disorders. A recent study suggesting a combination of appropriate probiotics and prebiotics through its ability to fix dysbiosis issues has emerged as a potential alternative to treat various pathological conditions of the Gastrointestinal Tract (GIT). Evidenced-based clinical research suggests that probiotic's effectiveness depends on several factors including strain, dose, host immune system, underlying pathology, and duration of therapy. Moreover, the microflora enriched medium reinforces the host defense against chemical and microbial challenges. However, the limited information available to understand the molecular basis by which probiotic maintains the gut homeostasis. The objective of this review to emphasize recent clinical outcomes and possible mechanisms involved in probiotic action to combat the GIT disorders.

Since its origin in the Wuhan province of China in December 2019, Coronavirus Disease 19 (COVID-19) has spread to most parts of the world and has infected millions of people. However, the significant variability in the mortality rate across the world indicates some underlying factors, especially the immunity factors that may have a potential role in this variability. One such factor that is being discussed and tested is the Bacillus Calmette-Guerin (BCG) vaccine. The available evidence suggests that BCG vaccination provides broad protection against respiratory infections as well as other infections. Therefore, BCG may prove to be a barrier for COVID-19 infection and may offer a ray of hope. In this review, we contrasted BCG vaccination program with COVID-19 mortality and analyzed trained immunity and cross protection against unrelated pathogens due to BCG vaccination. On analyzing the available data, we observed that countries without universal BCG vaccination policy are severely affected, while countries having universal BCG policies are less affected. Based on these data, we propose that the SARS-CoV-2 related qualified immunity, cross protection against unrelated pathogens and COVID-19 impact variations could be partly explained by the different national policies regarding BCG childhood vaccination. The combination of reduced morbidity and mortality may make BCG vaccination a potential new tool in the fight against COVID-19.

N-Acetylcysteine (NAC) has been proposed to be used to treat Coronavirus Disease 2019 (COVID-19). By reviewing the existing pathological studies of COVID-19, it was found that abundant mucus secretion, formation of a hyaline membrane (supportive of acute respiratory distress syndrome), and interstitial fibrous exudation may be important characteristics of COVID-19 and pathological targets of drug therapy. In addition, multiple extrapulmonary organ injuries in COVID- 19 may be associated with cytokine storm. NAC is an important antioxidant and anti-inflammatory drug. NAC has been demonstrated to have mucolytic effects in bronchitis, relieve respiratory failure in acute respiratory distress syndrome, and inhibit fibrous exudation in interstitial lung disease in clinical studies. These findings suggest that NAC may have a therapeutic effect on the pathological targets of COVID-19. Furthermore, NAC decreases TNF-α, IL-1β, IL-6, IL-8, IL-10, and IL-17 serum levels in patients with sepsis, severe burns, acute liver failure, or peritoneal dialysis and may also reduce cytokine storm in COVID-19. The antiviral effect of NAC on other respiratory viruses may also benefit COVID-19 patients. Summarizing the potential mechanisms of NAC in treating COVID-19 suggests that the role of NAC in COVID-19 treatment is worthy of further research.

Coronavirus-19 is a severe acute respiratory disorder in humans which has become a major health problem. It spreads out very rapidly throughout the world since it has been first identified in Wuhan, China (December 2019). The causative virus is known as severe acute respiratory syndrome coronavirus 2. And the World Health Organization has named this respiratory syndrome as a new epidemic disease called COVID-19. The incidence of COVID-19 continued to increase with three million confirmed infected cases and with 244,000 death cases worldwide. Until now there is no specific treatment or vaccine available against COVID- 19. The collective information about the different aspects of COVID-19 viral infection has been gathered from renowned journals, and electronic databases including Science Direct, Web of Science, Scopus and PubMed from 1990 to 2020. This manuscript has highlighted the transmission and symptoms of Covid-19. Therefore, these studies show how the SARS-CoV 2 can facilitate the debut of the virus into targeted host cells.

Objective: The purpose of this study was to explore the effect of TRAF1 on phenotypic changes of KCs in I/R in liver transplantation. Methods: SD rats were randomly divided into sham group and liver transplantation I/R group. KCs were extracted from rat livers in each group. KCs were transfected by lentivirus of si-TRAF1 or si-HOIP, and induced by Lipopolysaccharide (LPS). Flow cytometry was used to detect cell apoptosis. Western blot and RT-PCR were used to detect the protein and mRNA expression levels. Results: Compared with the sham group, the expression levels of TRAF1, TNF-α and IL-1β were significantly increased in the I/R group (P<0.05). In addition, compared with the control group, the expression levels of NF-ΚB (P65 and p-P65) and M1 phenotype (TNF-α and IL-1β) were notably increased in si-TRAF1 and si-HOIP group (all P<0.05). Furthermore, the levels of Linear Ubiquitin Complex (LUBAC) were markedly increased in LPS-induced KCs in comparison with the control group (P<0.05). Moreover, compared with the control group, the expression levels of P65, p-P65, TNF-α and IL-1β were notably decreased in the si-TRAF1 and si-HOIP group (P<0.05). The expression levels of P65, p-P65, TNF-α and IL-1β were significantly increased in si-TRAF1 and si-HOIP group when compared to the control group (P<0.05). Conclusion: TRAF1 was an important negative regulator of liver transplantation I/R by inhibiting the activation of NF-ΚB in KCs and preventing its M1 phenotype transformation.

Background: Amygdalin has anticancer benefits because of its active component, hydrocyanic acid. However, the underlying molecular mechanism is unclear. Objective: This study aimed to investigate the molecular mechanism by which amygdalin exerts antiproliferative effects in the human Michigan Cancer Foundation-7 (MCF-7) breast cancer cell line. Methods: MCF-7 cells were exposed to amygdalin at a particular IC50 value for 24 and 48 hours and compared to non-treated cells. An Affymetrix whole-transcript expression array was used to analyze the expression of 32 genes related to DNA replication. Results: Among the 32 genes, amygdalin downregulated the expression of 16 genes and 19 genes by >1.5-fold at 24 and 48 hours, respectively. At 24 hours, the downregulated genes from the DNA polymerase α-primase complex were POLA1, POLA2, PRIM1, and PRIM2; DNA polymerase δ complex: POLD3; DNA polymerase ε complex: POLE4, Minichromosome Maintenance protein (MCM) complex (helicase): MCM2, MCM3, MCM4, MCM6, and MCM7; clamp and clamp loader: PCNA; nuclease: FEN1; and DNA ligase: LIG1. At 48 hours, the downregulated genes from the DNA polymerase α-primase complex were POLA1, POLA2, and PRIM1; DNA polymerase δ complex: POLD3; DNA polymerase ε complex: POLE and POLE2; MCM complex (helicase): MCM2, MCM3, MCM4, MCM5, MCM6, and MCM7; clamp and clamp loader: PCNA, RFC2, and RFC3; RNase H: RNASEH2A; nucleases: DNA2 and FEN1; and DNA ligase: LIG1. Conclusion: Amygdalin treatment caused downregulation of several genes that play critical roles in DNA replication in the MCF-7 cell line. Thus, it might be useful as an anticancer agent.

Aim: To study the bactericidal activity of crude ethanolic extract and fractionations obtained from Sargassum aquifolium (Turner) C. Agardh (brown algae) towards Gram-positive bacteria and Gram-negative biofilm-forming human pathogenic bacteria. Background: The increasing emergence of antibiotic-resistant bacteria in the hospital and community settings has led to the discovery of alternative strategies. Marine organisms are considered as one of the potential sources of diverse bioactive molecules against several biological activities. Hence, the algae, especially the marine brown algae were selected to evaluate its antibacterial activities towards biofilm-forming human pathogenic bacteria. Objective: To restrain the drug-resistant ability of pathogenic bacteria, we checked the extract of Sargassum aquifolium (Turner) C. Agardh (Phyophyceae) for the concerned bioactive compounds. Methods: Antibacterial activity towards both Gram-positive and Gram-negative bacteria was evaluated using disk diffusion and broth microdilution assays. Furthermore, the active compound present in the extracts was also identified using Gas-Chromatography-Mass Spectroscopy (GC-MS). Results: A total of 21 bioactive compounds were identified using GC-MS analysis with different chemical natures. The crude ethanolic extraction was fractionated sequentially according to the eluotropic series from less to extreme polar. The highest zone of inhibition was recorded for ethanolic extract on Listeria monocytogenes with a value of 38.00±0.17 mm and the lowest was 10.67±0.06 mm for ethyl acetate fraction on Pseudomonas aeruginosa. Ethyl acetate fractionate showed a higher effectivity than other fractionations. An MIC value of 256 μg/mL was recorded against Staphylococcus aureus and L. monocytogenes and 512 μg/mL against Escherichia coli and P. aeruginosa. Its ethanolic extract also showed synergism with oxytetracycline on S. aureus, L. monocytogenes, and E. coli. Furthermore, the same extracts also showed synergism with tetracycline on E. coli and with erythromycin on P. aeruginosa. Conclusion: The present study reports the antibacterial activity of the S. aquifolium (Turner) C. Agardh extracts against human pathogenic bacteria. Furthermore, it also predicts the synergistic activity of selected antibiotic combinations against both selected Gram-positive and Gram-negative pathogenic bacteria.

Background: Lately, the employment of nano-carriers has been known as an optimistic means of drug and vaccine delivery. Objective: Nano vaccines are a novel tier of vaccines that can develop humoral and cellular immune responses and can be introduced as a practical and secure nano vaccine candidate for the prevention of diseases. The purpose of this study was to accomplish and use biodegradable nano-carriers for the synthesis of pentavalent vaccine and immunogenicity evaluation in the animal models. The PLGA nanoparticle was prepared and modified with chitosan nanoparticles. Nano-carrier PLGA-chitosan was loaded by the DTP-HepB-Hib antigens and confirmed by DLS, SEM, TEM and FT-IR, then in vitro loading and release were evaluated. Toxicity was assessed by the MTT method in the Hec293 cells. Antigenicity evaluation and histopathological study were conducted by injection of new nano pentavalent vaccines in the BALB/c mice and the immune response was measured in the serum samples through an indirect ELISA method. Results: Conclusions drawn from the current study exhibited the plausible ability of nano-carrier to deliver the vaccine. A notable increase was shown in total IgG and IgM antibodies examined in mice vaccinated with new nano vaccines. The histopathological study in the treated mice showed no toxicity in the vital organs of mice. Conclusion: The engineered vaccine delivery system showed the ability to induce robust immune responses and also suitability features of the PLGA-chitosan as a promising carrier to improve vaccine delivery.

Background: Pulmonary arterial Hypertension (PH) is a chronic disease that ultimately progresses to right ventricular failure and death. Until now, there is still a lack of effective treatment applied. The purpose of the present study was to observe the protective effect of Mesenchymal Stromal Cell-Derived Exosomes (MSC-EXO) against experimental Pulmonary arterial Hypertension (PH) and right ventricular failure. Methods: All the experimental rats received an intraperitoneal injection of 50 mg/kg monocrotaline to induce PH model. Three weeks after the model was successfully established, the cell Culture Media (CM) or MSC-EXO derived from human umbilical cord was administered daily via the tail vein. All animals were randomly divided into 4 groups: Control (saline-treated), MCT-PH, MCT-CM and MCT-EXO groups. Post-operation, hemodynamic data and index of right ventricular hypertrophy (RVHI) were recorded to evaluate the inhibition of MSC-EXO on MCT-induced PH. Histology, immunohistochemistry and western blot were used to analyze the effect of MSC-EXO against vascular remodeling and further reveal the mechanism. Results: In the present study, our results showed that MSC-EXO administration could significantly reduce the Right Ventricular Systolic Pressure (RVSP) and RVHI, suppress the pulmonary vascular remodeling and The Endothelial-Mesenchymal Transition (EndMT) process. Conclusion: Our results provided the firm information for a new method in the treatment of PH; the mechanism may be related to the inhibition of vascular remodeling and EndMT.

Background: Epithelial-Mesenchymal Transition (EMT) is a molecular reprogramming that leads to an increased ability to migrate, which can promote invasion and metastasis. EMT can be initiated in response to the activity of signaling pathways such as Wnt as well as miRNAs. Objective: The aim of the study was to determine the expression profile of EMT-related genes involved in signal transduction via the Wnt pathway and cadherins and to assess which miRNAs can participate in the regulation of their expression. Methods: The study material consisted of 50 endometrial samples: 40 with diagnosed endometrial cancer and 10 without neoplastic changes. The expression profile of EMT-related genes was assessed with microarrays and validated by RT-qPCR. MicroRNA expression profiling was performed using microarrays. It was also determined which miRNAs may participate in the expression regulation of EMT-related genes. Results: CDH1 overexpression was observed in all three endometrial cancer grades using both mRNA microarrays and RT-qPCR. The microarray experiment showed a decrease in CDH2 level regardless of the endometrial cancer grade, however, it was only partially validated with RT-qPCR. Low levels of WNT2, WNT4, WNT5A have also been observed. Decreased expression of WNT2 and WNT5A may be caused by miR-331-3p and miR-200b-5p, respectively. Conclusion: The Wnt signaling is disrupted in endometrial cancer, which may be due to miR-331- 3p and miR-200b-5p activity. In addition, a change in WNT5A level in endometrial cancer compared to control may indicate that it acts as a suppressor gene and that its low expression is associated with tumor progression.

Background: The pathogenesis of Severe Acute Pancreatitis (SAP) is mediated substantially by dysfunctions in the intestinal barrier. Euphorbia kansui (EK) is a medicinal plant used widely in traditional Chinese medicine to treat inflammation; however, its efficacy and mechanism of action in SAP treatment are not yet well understood. Objective: To investigate the role of EK in intestinal barrier tissue repair and in the pathogenesis and development of SAP. Methods: The rat SAP model was established by a retrograde injection of sodium taurocholate into the pancreatic bile duct. The SAP model group and the SAP + EK treatment groups were divided into 6 subgroups according to timing: 2, 6, 12, 24, 48, or 72h after inducing SAP. The progression of the SAP rats and of the rats receiving the EK treatment was evaluated using the ascites volume, serum amylase and plasma endotoxin levels, and histological grading of intestinal mucosal damage. In addition, serum inflammatory factor contents were measured using Enzyme-Linked Immunosorbent Assay (ELISA) tests and apoptotic cells in damaged ileum tissue were detected using TUNEL staining. Apoptosis markers and other signaling proteins in intestinal mucosal cells were detected by immunohistochemical assays and then validated by combining these data with quantitative polymerase chain reactions and western blotting. Results: Compared with the results of the SAP model rats, the results of the rats that received EK treatment demonstrated that EK could effectively reduce the ascites volume and serum amylase and plasma endotoxin levels. EK treatment also greatly reduced the abnormal intestinal morphological alterations in the rat SAP model and significantly downregulated the serum contents of Interleukin (IL)-1β, IL-6, and tumor necrosis factor-α. EK treatment inhibited the elevation of capapse-3, inhibited the decrease of the Bcl-2 protein, and decreased the number of apoptotic cells in rat ileum tissue. Finally, EK treatment abrogated the increase of HMGB1 and the suppression of MFG-E8 protein expression in the SAP + EK rat ileum tissue. Conclusion: EK suppresses SAP pathogenesis by restoring the intestinal barrier function and modulating the HMGB1/MFG-E8 signaling axis.