Clinical and Experimental Pharmacology and Physiology最新文献

The integrity of the Blood-Brain Barrier (BBB) is crucial in the pathophysiological progression of acute ischemic stroke (AIS). However, the potential of Kisspeptin-10 (Kp-10), with its antioxidant and anti-inflammatory properties, has not been explored experimentally in the context of stroke management. This study aimed to investigate the neurovascular protective effects of Kp-10 following cerebral ischemia using both in vivo and in vitro models. A middle cerebral artery occlusion (MCAO) model was established in C57BL/6 mice, followed by Kp-10 administration. Neurological deficits (Longa score), infarct volume (TTC staining), BBB permeability (14C-Sucrose), and Claudin-10 expression (qRT-PCR and immunohistochemistry) were assessed to evaluate the therapeutic effects of Kp-10. Human brain microvascular endothelial cells (HBMVECs) were subjected to oxygen–glucose deprivation/reoxygenation (OGD/R) to mimic ischemic conditions. Endothelial permeability, oxidative stress (OS), and nuclear factor erythroid 2-related factor 2 (Nrf2) levels were evaluated. Nrf2 silencing was performed to validate its role in Kp-10-mediated protection. Initially, we observed a significant reduction in Kp-10 expression within the cortical tissue of mice subjected to MCAO. Subsequent administration of Kp-10 not only alleviated neurological deficits but also significantly mitigated blood–brain barrier (BBB) dysfunction following stroke induction, as evidenced by reduced 14C-sucrose leakage. Furthermore, Kp-10 treatment led to an upregulation of Claudin-10 expression in the post-stroke cortical region. In our in vitro experiments, we employed HBMVECs exposed to OGD/R to simulate ischemic conditions. We found that Kp-10 effectively reduced OGD/R-induced endothelial permeability by enhancing Claudin-10 expression. Additionally, Kp-10 exhibited antioxidant capabilities by decreasing mitochondrial reactive oxygen species (ROS) levels, increasing superoxide dismutase (SOD) activity, and upregulating nuclear factor erythroid 2-related factor 2 (Nrf2) expression. Notably, when Nrf2 was knocked down in HBMVECs, the protective effects of Kp-10 on endothelial permeability and Claudin-10 expression were abolished, indicating that the beneficial actions of Kp-10 are mediated through the Nrf2 pathway. In conclusion, our findings suggest that Kp-10 holds promise as a therapeutic strategy to preserve BBB integrity and promote neuroprotection following stroke, acting primarily via the Nrf2 signalling pathway. These findings suggest that Kp-10 may represent a promising therapeutic strategy for preserving BBB integrity following ischemic stroke.

Gestational diabetes mellitus (GDM) is a metabolic condition defined by glucose intolerance during pregnancy, which frequently results in placental malfunction, oxidative stress, and chronic inflammation, all of which are linked to the activation of the NLRP1 inflammasome. GPR39, a G protein-coupled receptor, has emerged as a possible therapeutic target, and its agonist, TC-G 1008, may provide protection against metabolic disorders. In this study, we investigated the expression of GPR39 in the murine placenta and its role in GDM-related placental dysfunction using a high-fat diet-induced GDM mouse model. Mice were treated with TC-G 1008 (7.5 and 15 mg/kg) from gestational day 0 to 18, and placental tissues were analysed via RT-PCR, Western blot, immunohistochemistry, ELISA, and biochemical assays. Our findings indicated that GPR39 expression was markedly reduced in GDM mice relative to wild-type controls. Administration of TC-G 1008 (7.5 and 15 mg/kg) enhanced fetal outcomes, including survival rate, weight, and crown-rump length, while also mitigating maternal metabolic disorders such as hyperglycaemia, insulin resistance, and dyslipidaemia. Moreover, the administration of TC-G 1008 mitigated placental oxidative stress by augmenting SOD activity and GSH levels, while inhibiting NLRP1 inflammasome activation, as shown by diminished expression of NLRP1, ASC, and Caspase-1, alongside lowered levels of IL-1β and IL-18. Furthermore, GPR39 activation suppressed the NF-κB and MAPK signalling pathways. These findings suggest that GPR39 activation by TC-G 1008 ameliorates placental dysfunction in GDM by mitigating oxidative stress and inflammation via suppression of the NLRP1 inflammasome, highlighting its potential as a therapeutic strategy for GDM-associated complications.

Psoriasis is a complex inflammatory autoimmune skin illness that causes epidermal keratinocyte hyperproliferation and dedifferentiation. In a previous study we did in the lab, we found that treating human keratinocytes (HaCaT) with PUVA increased the expression of hsa-miR-718 compared to control. In this study, an imiquimod (IMQ)-induced mouse model and HaCaT were used to look at how overexpressing miR-718 could help treat psoriasis and its causes. To gain more understanding, the in vivo study used the JAK1/3 inhibitor tofacitinib. We observed that miR-718 overexpression leads to the inhibition of JAK–STAT signalling, as evidenced by the reduced expression of STAT1, JAK proteins, and their phosphorylated forms, both in vitro and in vivo. Luciferase assay demonstrated the direct inhibition of STAT1 due to miR-718 overexpression. Additionally, in vitro experiments showed downregulation of STAT2 and STAT3. Inhibition of basal miR-718 in HaCaT overactivates JAK–STAT signalling proteins. In psoriatic mice, ectopic miR-718 reduces NF-kB, a key mediator of inflammation. IHC shows lower acanthosis and parakeratosis in IMQ-induced psoriatic mice. In the skin of mice that had been miR-718 transfected, there was less VEGF, MMP7 and MMP9. Understanding how miR-718 improves psoriasis not only provides new information but also inspires hope for its potential use as a psoriasis treatment.

Sepsis is the leading cause of acute lung injury (ALI), and kaempferol has a protective effect against ALI. However, the underlying mechanisms have not been fully elucidated. This study aimed to investigate the role of kaempferol in sepsis-induced ALI and its underlying molecular mechanism, particularly the involvement of succinylation. Human pulmonary microvascular endothelial cells were treated with lipopolysaccharide to induce injury. Cell apoptosis and inflammation were evaluated by flow cytometry and enzyme-linked immunosorbent assay. Succinylation was analysed using immunoblotting, co-immunoprecipitation and protein stability assay. The results showed that kaempferol inhibited apoptosis and inflammation response in LPS-treated HPMVECs and attenuated lung damage in septic mice. Moreover, kaempferol enhanced succinylation levels and reduced SIRT5 protein levels. SIRT5 suppressed the succinylation of SRPK1 at lysine (K) 588 site and facilitated SRPK1 degradation. Overexpression of SIRT5 or knockdown of SRPK1 reversed the inhibitory effects of kaempferol or SIRT5 knockdown on cellular biological behaviours, respectively. In conclusion, kaempferol attenuates sepsis-induced lung injury by inhibiting HPMVEC apoptosis and inflammation. Mechanistically, kaempferol suppresses SIRT5-mediated desuccinylation of SRPK1, thereby promoting SRPK1 protein stability. The findings suggest the important role of SRPK1 succinylation in ALI and kaempferol may be used for the treatment of the disease.

Whole blood hydroxychloroquine (WBHCQ) levels were found to correlate with cutaneous lupus (CLE) disease severity. Studies have shown HCQ to be highly variable in blood concentrations and clinical response among patients; however, the exact cause of this variation is not well understood. Genetic polymorphism of CYP450 enzymes was suspected to be a possible contributing factor. This study aimed to determine the effects of genetic polymorphism of CYP450 enzymes on clinical response to HCQ among Malaysians with CLE and its association with blood [HCQ], [DHCQ] or [HCQ]: [DHCQ] ratio. This study was a multicentre cohort study targeting CLE subjects on HCQ, recruited from five main hospitals in Malaysia. A total of 33 subjects were recruited from 1 September 2021 to 30 November 2023. Blood [HCQ], [DHCQ] or its concentration ratio and clinical response to HCQ were the outcome variables determined. Statistical Analysis showed that CYP2D6*10 was significantly associated with blood [HCQ] (p < 0.05), while CYP3A5*3 was shown to be significantly associated with [DHCQ]: [HCQ] ratio (p < 0.05). However, a statistically significant association was not observed between identified SNPs and clinical response to HCQ (OR: 4.444, CI: 0.770, 25.654, p = 0.095). These findings are aligned with those reported previously on blood HCQ and CYP polymorphism; however, clinical response to HCQ and CYP polymorphism is still debatable, as we did not observe any significant association. CYP450 genetic polymorphism significantly affects levels of HCQ in patients. Thus, individual genotypes of CYP450 enzymes may be considered for the optimum use of HCQ among CLE subjects.

Immune checkpoint inhibitors (ICIs) targeting programmed cell death-1 (PD-1) and programmed cell death ligand-1 (PD-L1) have improved cancer outcomes but can induce immune-related adverse events, including mycobacterial infections. This study retrospectively analysed pulmonary nontuberculous mycobacteria (NTM) infection in 91 patients with pre-existing stage II–IV lung cancer at Shanghai Pulmonary Hospital between June 2015 and June 2024. Among these patients, 78.0% [71/91] were male, and 16.5% [15/91] were receiving ICIs monotherapy at the time of initial pulmonary NTM infection diagnosis (ICIs group). Compared with the non-ICIs group, ICIs-treated patients developed pulmonary NTM infection earlier (median time: 12 months [IQR, 6–18] vs. 21.5 months [IQR, 13–30]; p = 0.004), and had a higher proportion of rapid-growing NTM isolates (73.3% [11/15] vs. 30.3% [23/76]; p = 0.003), predominantly Mycobacterium abscessus complex (40.0% [6/15]). In contrast, Mycobacterium avium complex was the predominant NTM species in the non-ICIs group (71.1% [54/76]). During 12 months of anti-NTM therapy, the ICIs group showed a lower cumulative sputum culture conversion rate (36.4% [4/11] vs. 59.6% [31/52]; p = 0.19) and a longer median time to initial sputum culture conversion (12 months vs. 6 months; p = 0.13), though these differences were not statistically significant. The most commonly administered ICIs drugs were tislelizumab (40.0% [6/15]) and sintilimab (26.7% [4/15]). These findings suggest that ICIs may be associated with earlier development of pulmonary NTM infection in lung cancer patients. Future prospective studies with larger sample sizes are needed to validate this conclusion.