Inhibition of Rac 1 Protect Against Platelet Induced Liver and Kidney ‎Injury in Diabetes Mellitus

Diabetes mellitus both (Type 1 and Type 2) are one of the common causes for activation of platelet. Inflammation-induced abnormal platelet function contributes to chronic complications, which are the leading causes of death and morbidity among diabetics. Rac1 has been shown to regulate a variety of platelet functions; predicted Rac1could regulate platelet release of CXCL4, which leads to kidney injury in Diabetes Mellitus. Diabetes mellitus' effect on Rac1 activation, a 21kD G-protein implicated in platelet activation, was investigated and platelet induced inflammation and kidney injury. Swiss albino male mice were pretreated with 5 mg/kg of a specific Rac1 inhibitor NSC23766 and injected with (45 mg/kg body wt.) streptozotocin, twice for five days. Moreover, the concentration of serum chemokines CXCL4 were assayed using ELISA and histology score for kidney were examined. Our results showed that Diabetes mellitus was induced in mice by streptozotocin. In addition, platelet chemokines (CXCL4) were markedly higher in diabetic mice when compared to the sham (control) group. Moreover, pretreatment with NSC23766 decreased liver and kidney injury assessed by histology score, P-value <0.05. Our study reveals that Rac1 has a critical role in platelet chemokines secretion due to diabetes-induced inflammation in the liver and kidneys, targeting Rac1 could be a target for innovative treatment to control inflammation in diabetic individual. Targeting platelets involved in inflammatory pathways could be part of a strategy in order to control and manage diabetes and its consequences

Rac1 inhibition protect against platelet induced organ injury in Diabetes Mellitus

Background: Diabetes mellitus is one of the common causes for activation of platelet. Inflammation-induced abnormal platelet function contributes to chronic complications, which are consider the leading causes of death and morbidity among diabetics. Objective: Rac1, a 21kD G-protein has been shown to regulate a variety of platelet functions; we predicted that Rac1 could regulate platelet release of CXCL4 and CCL5, which may leads to organ injury in Diabetes Mellitus. Patients and Methods: Diabetes Mellitus' effect on Rac1 activation implicated in platelet activation, was investigated as platelet-induced inflammation and organ injuries. Swiss albino male mice were pretreated with 5 mg/kg of a specific Rac1 inhibitor NSC23766 and injected with (45 mg/kg body wt.) streptozotocin, twice for five days. Moreover, the concentration of serum chemokines CXCL4 and CCL5 were assayed using ELISA, and histology scores for kidney and pancreas was examined. Results: Our results show that Diabetes Mellitus was induced in mice by streptozotocin. In addition, platelet chemokines (CXCL4, CCL5) were markedly higher in diabetic mice when compared to the sham group. Moreover, pretreatment of diabetic mice induced by STZ, with NSC23766 decreased kidney and pancreatic injuries assessed by histology score, P-value <0.05. Conclusion: Our study reveals that Rac1 has a critical role in platelet chemokines secretion due to diabetes-induced inflammation in the kidneys and pancreas, and targeting Rac1 could be a target for innovative treatment to control inflammation in a diabetic individual. Targeting platelets involved in inflammatory pathways could be part of a strategy in order to control and manage diabetes consequences

Analysis of serum lipid profile in adult female smokers in Erbil city, Kurdistan region of Iraq

Background: There are few studies demonstrated the association between smoking and lipid profile in female adult smokers. Materials and Methods: This study conducted to determine and compare the serum lipid profile of female adult smokers with non-female smokers, known as controls. In 180 female subjects, the level of serum lipid profile measured. Results: Of these, 110 were smokers and 70 non-smokers (control) aged between 25 and 50 years. The study involved only smokers who had smoked for more than 5 years. Our result revealed that mean serum of total cholesterol (275.2 ± 32.6 mg/dl), triacylglycerol (188.4 ± 56.42 mg/dl), very low density lipoprotein (36.6 ± 14.2 mg/dl), low density lipoprotein (133.21 ± 9.81 mg/dl) were significantly higher in female smokers as compared to non-female smokers with mean of serum total cholesterol (172.3 ± 18.6 mg/dl), very low density lipoprotein (21.8 ± 9.6 mg/dl), triglyceride (108 ± 8.84 mg/dl), low density lipoprotein (94.54 ± 8.5 mg/dl). However, the mean of serum value for high density lipoprotein cholesterol in chronic female smokers was lower (44.6 ± 4.6 mg/dl) than in non-female smokers (55.3 ± 8.2 mg/dl). Conclusion: This study indicated that smoking cigarettes in female cause’s dyslipidemia, resulting in increased risk of cardiovascular disease among smokers

Analysis of cytokines in SARS-CoV-2 or COVID-19 patients in Erbil city, Kurdistan Region of Iraq

The emergence of the novel coronavirus and then pandemic outbreak was coined 2019- nCoV or COVID-19 (or SARS-CoV-2 disease 2019). This disease has a mortality rate of about 3·7 percent, and successful therapy is desperately needed to combat it. The exact cellular mechanisms of COVID-19 need to be illustrated in detail. This study aimed to evaluate serum cytokines in COVID-19 patients. In this study, serum was collected from volunteer individuals, moderate COVID-19 patients, severe cases of COVID-19 patients, and patients who recovered from COVID-19 (n = 122). The serum concentrations of interleukins such as IL-1, IL-4, IL-6, IL-8, IL-10, and tumor necrosis factor-alpha (TNF-α), were measured by enzyme-linked immunosorbent assays (ELISA). The concentrations of IL-1 and TNF-α were did not differ significantly among groups. However, the concentration of IL-6 was significantly higher in moderate COVID-19 and severe cases of COVID-19 groups compared to control and recovered groups indicating it to be an independent predictor in the coronavirus disease. The levels of IFN-γ and IL-4 were significantly lower in the recovery group than the severe case of the COVID-19 group. In contrast, the level of IL-10 in recovered COVID-19 patients was significantly higher in compare to severe cases, COVID-19 patients. Varying levels of cytokines were detected in COVID-19 group than control group suggesting distinct immunoregulatory mechanisms involved in COVID-19 pathogenesis. However, additional investigations are needed to be to be performed to understand the exact cellular mechanism of this disease

Rac1 signaling regulates platelet-dependent inflammation abdominal sepsis

Sepsis is a systemic inflammatory response syndrome to a localized or systemic infection that leads to the over production of proinflammatory cytokines and the ultimate failure of multiple organ systems. However, little is known about the potential role of Rac1 in controlling sepsis-induced intracellular signaling pathways. We hypothesized that Rac1 might be involved in sepsis mediated signaling pathways leading to the activation of inflammatory cells. Paper (I), Rac1 signaling plays an important role in polymicrobial sepsis induced by cecal ligation and puncture (CLP). This study shows that Rac1 signaling regulates sepsis-induced inflammation in the lung by reducing chemokine production and Mac-1 expression on neutrophils. Rac1 inhibitor NSC23766 attenuates lung edema, tissue destruction and systemic pro-inflammatory cytokines in septic animals, suggesting that targeting Rac1 may be useful approach to protect against pulmonary injury in abdominal sepsis. Paper (II), in this study we showed that inhibition of Rac1 signaling protect sepsis-induced lung injury through two different mechanisms. First, Rac1 controls surface mobilization of CD40L on activated platelets and second, Rac1 regulates MMP-9 secretion from neutrophils. Our data indicate that inhibition of Rac1 signaling might be a useful target in order to control pathological secretion and shedding of CD40L into the systemic circulation in abdominal sepsis. Paper (III) This study indicates that Rac1 activity is increased in platelets and regulates platelet secretion of CCL5 in abdominal sepsis. Our findings show that CCL5 regulates neutrophil recruitment in septic lung injury via activation of alveolar macrophages leading to local secretion of CXCL2. Thus, our novel data not only elucidates complex mechanisms regulating pulmonary neutrophil trafficking in sepsis but also suggest that targeting Rac1 signaling and platelet-derived CCL5 might be a useful way to control pathological inflammation and tissue damage in the lung in abdominal sepsis. Paper (IV) Rac1 signaling is enhanced in platelets and regulates platelet secretion of CXCL4 in abdominal sepsis. This study indicates that CXCL4 controls neutrophil accumulation via secretion of CXCL2 from alveolar macrophages in septic lung injury. These findings not only delineate complex mechanisms of neutrophil trafficking in sepsis but also suggest that targeting platelet-derived CXCL4 might be an effective way to ameliorate inflammation and tissue damage in septic lung damage

Platelet secretion of CXCL4 is Rac1-dependent and regulates neutrophil infiltration and tissue damage in septic lung damage.

Platelets are potent regulators of neutrophil accumulation in septic lung damage. We hypothesized that platelet-derived CXCL4 might support pulmonary neutrophilia in a murine model of abdominal sepsis

Rac1 regulates platelet shedding of CD40L in abdominal sepsis.

Matrix metalloproteinase-9 (MMP-9) regulates platelet shedding of CD40L in abdominal sepsis. However, the signaling mechanisms controlling sepsis-induced shedding of CD40L from activated platelets remain elusive. Rac1 has been reported to regulate diverse functions in platelets; we hypothesized herein that Rac1 might regulate platelet shedding of CD40L in sepsis. The specific Rac1 inhibitor NSC23766 (N6-[2-[[4-(diethylamino)-1-methylbutyl] amino]-6-methyl-4-pyrimidinyl]-2 methyl-4, 6-quinolinediamine trihydrochloride) was administered to mice undergoing cecal ligation and puncture (CLP). Levels of CD40L and MMP-9 in plasma, platelets, and neutrophils were determined by use of ELISA, western blot, and confocal microscopy. Platelet depletion abolished the CLP-induced increase in plasma levels of CD40L. Rac1 activity was significantly increased in platelets from septic animals. Administration of NSC23766 abolished the CLP-induced enhancement of soluble CD40L levels in the plasma. Moreover, Rac1 inhibition completely inhibited proteinase-activated receptor-4-induced surface mobilization and secretion of CD40L in isolated platelets. CLP significantly increased plasma levels of MMP-9 and Rac1 activity in neutrophils. Treatment with NSC23766 markedly attenuated MMP-9 levels in the plasma from septic mice. In addition, Rac1 inhibition abolished chemokine-induced secretion of MMP-9 from isolated neutrophils. Finally, platelet shedding of CD40L was significantly reduced in response to stimulation with supernatants from activated MMP-9-deficient neutrophils compared with supernatants from wild-type neutrophils, indicating a direct role of neutrophil-derived MMP-9 in regulating platelet shedding of CD40L. Our novel data suggest that sepsis-induced platelet shedding of CD40L is dependent on Rac1 signaling. Rac1 controls surface mobilization of CD40L on activated platelets and MMP-9 secretion from neutrophils. Thus, our findings indicate that targeting Rac1 signaling might be a useful way to control pathologic elevations of CD40L in the systemic circulation in abdominal sepsis.Laboratory Investigation advance online publication, 21 July 2014; doi:10.1038/labinvest.2014.92

Rac1-dependent secretion of platelet-derived CCL5 regulates neutrophil recruitment via activation of alveolar macrophages in septic lung injury.

Accumulating evidence suggest that platelets play an important role in regulating neutrophil recruitment in septic lung injury. Herein, we hypothesized that platelet-derived CCL5 might facilitate sepsis-induced neutrophil accumulation in the lung. Abdominal sepsis was induced by CLP in C57BL/6 mice. CLP increased plasma levels of CCL5. Platelet depletion and treatment with the Rac1 inhibitor NSC23766 markedly reduced CCL5 in the plasma of septic mice. Moreover, Rac1 inhibition completely inhibited proteasePAR4-induced secretion of CCL5 in isolated platelets. Immunoneutralization of CCL5 decreased CLP-induced neutrophil infiltration, edema formation, and tissue injury in the lung. However, inhibition of CCL5 function had no effect on CLP-induced expression of Mac-1 on neutrophils. The blocking of CCL5 decreased plasma and lung levels of CXCL1 and CXCL2 in septic animals. CCL5 had no effect on neutrophil chemotaxis in vitro, suggesting an indirect effect of CCL5 on neutrophil recruitment. Intratracheal challenge with CCL5 increased accumulation of neutrophils and formation of CXCL2 in the lung. Administration of the CXCR2 antagonist SB225002 abolished CCL5-induced pulmonary recruitment of neutrophils. Isolated alveolar macrophages expressed significant levels of the CCL5 receptors CCR1 and CCR5. In addition, CCL5 triggered significant secretion of CXCL2 from isolated alveolar macrophages. Notably, intratracheal administration of clodronate not only depleted mice of alveolar macrophages but also abolished CCL5-induced formation of CXCL2 in the lung. Taken together, our findings suggest that Rac1 regulates platelet secretion of CCL5 and that CCL5 is a potent inducer of neutrophil recruitment in septic lung injury via formation of CXCL2 in alveolar macrophages

Ras regulates alveolar macrophage formation of CXC chemokines and neutrophil activation in streptococcal M1 protein-induced lung injury.

Streptococcal toxic shock syndrome (STSS) is associated with a high mortality rate. The M1 serotype of Streptococcus pyogenes is most frequently associated with STSS. Herein, we examined the role of Ras signaling in M1 protein-induced lung injury. Male C57BL/6 mice received the Ras inhibitor (farnesylthiosalicylic acid, FTS) prior to M1 protein challenge. Bronchoalveolar fluid and lung tissue were harvested for quantification of neutrophil recruitment, edema and CXC chemokine formation. Neutrophil expression of Mac-1 was quantified by use of flow cytometry. Quantitative RT-PCR was used to determine gene expression of CXC chemokines in alveolar macrophages. Administration of FTS reduced M1 protein-induced neutrophil recruitment, edema formation and tissue damage in the lung. M1 protein challenge increased Mac-1 expression on neutrophils and CXC chemokine levels in the lung. Inhibition of Ras activity decreased M1 protein-induced expression of Mac-1 on neutrophils and secretion of CXC chemokines in the lung. Moreover, FTS abolished M1 protein-provoked gene expression of CXC chemokines in alveolar macrophages. Ras inhibition decreased chemokine-mediated neutrophil migration in vitro. Taken together, our novel findings indicate that Ras signaling is a potent regulator of CXC chemokine formation and neutrophil infiltration in the lung. Thus, inhibition of Ras activity might be a useful way to antagonize streptococcal M1 protein-triggered acute lung injury

Rac1 regulates bacterial toxin-induced thrombin generation.

Systemic inflammatory response syndrome is associated with severe coagulopathy. The purpose of this study was to examine thrombin generation in systemic inflammation triggered by the endotoxin lipopolysaccharide (LPS) and the exotoxin streptococcal M1 protein