Pakistan Journal of Pharmaceutical Sciences

Abstract: Background: Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by synovial inflammation, joint swelling, pain, cartilage destruction and bone erosion. Qingteng Waifu San is an external sinomenine-containing preparation, but its site-specific transdermal response requires disease-relevant evaluation. Objectives: To compare Zusanli (ST36)-site and adjacent non-acupoint transdermal application of Qingteng Waifu San in an ovalbumin/complete Freund’s adjuvant-induced RA rabbit model and to evaluate formulation, neuropeptide, inflammatory, behavioural and histopathological outcomes. Methods: Forty-eight New Zealand white rabbits were allocated into six groups (n=8 each): normal control, vehicle-only transdermal control, acupuncture, acupoint injection, ST36-site transdermal Qingteng Waifu San and adjacent non-acupoint transdermal Qingteng Waifu San. Gel appearance, pH, viscosity, spreadability, sinomenine content and high-performance liquid chromatography (HPLC) fingerprint similarity were assessed. Substance P (SP), calcitonin gene-related peptide (CGRP), tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and interleukin-1 beta (IL-1beta) were measured by enzyme-linked immunosorbent assay (ELISA). SP and CGRP messenger RNA (mRNA) were analysed by real-time polymerase chain reaction (PCR). Arthritis score, paw volume, mechanical withdrawal threshold and histopathological scores were assessed. Results: The vehicle-only group showed RA-related changes compared with normal controls. ST36-site Qingteng Waifu San reduced arthritis score, paw volume, serum cytokines and histopathological injury scores and improved mechanical withdrawal threshold compared with vehicle-only control. SP and CGRP levels were higher after ST36 application than after adjacent non-acupoint application, indicating site-dependent neurocutaneous modulation. Conclusion: ST36-site transdermal Qingteng Waifu San produced stronger neuropeptide modulation and more favourable RA-related outcome profiles than adjacent non-acupoint application.

Abstract: Background: Intravenous lidocaine reduces propofol requirements and procedure-related adverse events. Objectives: The study aimed to test whether intravenous lidocaine would reduce the effect-site concentration of propofol required to achieve loss of consciousness and decrease propofol consumption during total intravenous anesthesia in gynecological laparoscopy. Methods: This was a prospective, randomized, double-blind, placebo-controlled trial. Sixty patients were randomly allocated to receive either intravenous lidocaine (1.5 mg•kg-¹ bolus) followed by continuous infusion or an equal volume of saline. Propofol was administered via target-controlled infusion starting at an effect-site concentration of 3.5 μg/mL. The concentration was then adjusted in steps of 0.5 μg/mLaccording to Dixon’s up-and-down sequential method: decreased if loss of consciousness was achieved, or increased if not. Loss of consciousness was defined as loss of response to verbal commands. The median effective concentration (EC50) of propofol for inducing loss of consciousness was calculated using the Dixon’s up-and-down method. General anesthesia was maintained with propofol and remifentanil, guided by state entropy (target 40-60) and surgical pleth index (target 20-50). Drug consumption was normalized to anesthesia duration and body weight. Results: The estimated EC50 of propofol for inducing loss of consciousness was significantly lower in the lidocaine group than in the saline group (3.32 μg/mL, 95% Confidence Interval (CI): 3.04-3.59 vs. 3.89 μg/mL, 95% CI: 3.50-4.28). Under the study protocol, the lidocaine group also required less propofol (8.62 mg•kg-1•h-1, 95% CI: 8.10-9.15 vs. 9.89 mg•kg-1•h-1, 95% CI: 9.05-10.73) and less remifentanil (0.23 μg•kg-1•min-1, 95% CI: 0.21-0.24 vs. 0.27 μg•kg-1•min-1, 95% CI: 0.24-0.30) compared with the saline group. Conclusion: Intravenous lidocaine reduced the propofol EC50 for Loss of Consciousness (LOC) and decreased intraoperative propofol and remifentanil consumptions in patients undergoing gynecological laparoscopy. These findings suggest a propofol- and opioid-sparing effect of intravenous lidocaine in this setting, although confirmation in larger multicenter trials is needed.

Abstract: Background: Neuromyelitis optica spectrum disorders (NMOSD) are autoimmune demyelinating diseases of the central nervous system. The role of microRNA-34b (miR-34b) in NMOSD pathogenesis remains unclear. This study investigates the expression and functional mechanism of miR-34b in NMOSD, specifically its regulation of the TIA-1-stress granule pathway. Objectives: To determine the expression level of miR-34b in NMOSD tissues and cell lines and to elucidate whether miR-34b suppresses cell proliferation and promotes apoptosis by directly targeting the TIA-1-stress granule pathway. Methods: Real-time PCR was used to detect miR-34b expression in 25 NMOSD tissue specimens obtained from a hospital laboratory (13 early-stage, 12 chronic-stage) and 10 normal control tissues, as well as in RGC-5 cells. RGC-5 cells were transfected with miR-34b mimics, inhibitors or negative controls. Cell proliferation was assessed by MTT assay and cell cycle distribution and apoptosis was analyzed by flow cytometry (PI staining and Annexin V-FITC/PI double staining, respectively). The direct binding between miR-34b and the 3'-UTR of TIA-1 mRNA was validated using a dual-luciferase reporter assay. Protein expression levels of TIA-1, SG, HuR, CDK2 and BCL-2 were measured by Western blot. A rescue experiment was performed by co-transfecting miR-34b mimics with a TIA-1 overexpression plasmid. Results: miR-34b was lower in NMOSD vs controls (0.32 ± 0.08 vs. 1.00 ± 0.12, P<0.01), especially in the chronic stage. miR-34b mimics inhibited proliferation, induced G0/G1 arrest (68.5% vs. 52.1%, P<0.01) and increased apoptosis (22.6% vs. 8.3%, P<0.001). miR-34b inhibitor had opposite effects. miR-34b directly bound TIA-1 3'-UTR and downregulated TIA-1, SG, HuR. TIA-1 overexpression partially reversed miR-34b effects. Conclusions: miR-34b is downregulated in NMOSD. Overexpression of miR-34b suppresses proliferation and promotes apoptosis by targeting the TIA-1-stress granule pathway. Restoring miR-34b may be a therapeutic strategy for NMOSD.

Abstract: Background: The escalating prevalence of obesity has made it a critical public health concern. There is an urgent need to identify naturally derived compounds with anti-obesity potential. Britanin (BRI), a bioactive sesquiterpene lactone derived from Inula species, has shown promise in metabolic disorder management, but its anti-obesity mechanisms remain uncharacterized. Objectives: Combining animal experiments and network pharmacology analysis to explore the effect of BRI in high-fat diet-induced obesity. Methods: C57BL/6J male mice were used for experiment. A high-fat diet (HFD)-induced obese mouse model was treated with BRI (5/15 mg/kg, i.p.) to validate lipid metabolism and weight loss. Network pharmacology identified potential targets via SwissTargetPrediction, GeneCards and OMIM databases, with molecular docking (CB-DOCK) and PPI network analysis (STRING/Cytoscape). Relevant validations were conducted based on the screened targets. Additionally, a biosafety assessment was performed. Results: In-vivo, 15 mg/kg BRI reduced body weight by 18%, decreased serum TG (-45%, p<0.001), TC (-37%, p<0.001) and LDL-C (-32%, p<0.01) and reversed adipocyte hypertrophy. Thirty-nine intersection targets were identified, with MAPK1, EGFR, PTGS2, MAP2K1 and MAPK8 as top hubs (degree centrality >15). BRI exhibited strong binding affinity (-7.7 to -10.3 kcal/mol) to these targets. Mechanistically, BRI exerts its anti-obesity effects by regulating key targets within the MAPK signaling pathway, particularly MAPK1 and inhibited the PPARγ, thereby blocking adipogenesis and promoting the transition of adipose tissue. Conclusion: BRI may alleviate obesity by regulating the Mitogen-Activated Protein Kinase signaling pathway, providing a rationale for natural compound-based obesity therapy.

Abstract: Background: Diabetes retinopathy (DR) is the main blinding complication of type 2 diabetes (T2DM). Glycated hemoglobin (HbA1c) can reflect long-term blood glucose control and optical coherence tomography (OCT) can evaluate retinal microvessels. The effects of metformin and insulin regimens on microvascular structure are not yet clear. Objectives: To explore the relationship between HbA1c levels and retinal microvascular changes in T2DM patients receiving metformin and/or insulin and to evaluate the impact of different hypoglycemic regimens on DR progression. Methods: This retrospective study included T2DM patients with non proliferative DR treated between January 2023 and December 2024. After propensity score matching (PSM), 31 patients per group received metformin monotherapy (M), insulin based therapy (Ins), or combined therapy (M Ins). HbA1c, HbA1c variability (HbA1c SD), DR grade, central macular thickness (CMT), retinal nerve fiber layer (RNFL) thickness and superficial/deep capillary plexus (SCP/DCP) density were recorded at baseline, 1 and 2 year follow up. Multiple linear regression and Cox regression were used. Results: After 2 years, HbA1c and HbA1c SD decreased significantly in all groups (p<0.05), with greater reduction in the M Ins group. DR progression was documented in 23 patients (24.7%), with rates of 16.1%, 38.7% and 19.4% in the M, Ins and M-Ins groups, respectively (p>0.05). OCT showed reduced CMT/RNFL and increased SCP/DCP density in all groups (p<0.05), with better improvement in M Ins group (p<0.05). HbA1c correlated positively with CMT/RNFL and negatively with SCP/DCP (p<0.05). Cox regression indicated that higher HbA1c-SD was an independent risk factor for DR progression (HR = 3.216, 95% CI: 1.011–10.230, p= 0.048), while metformin use was an independent protective factor (HR=0.694, 95% CI: 0.451–0.840, p=0.001). Conclusion: Metformin plus insulin was associated with greater HbA1c reduction, decreased glycemic variability and increased retinal vessel density and may thus slow DR progression. Both HbA1c-SD and SCP density were correlated with DR progression risk, supporting the combined use of glycemic variability and OCT parameters for clinical evaluation and treatment guidance.