Pakistan Journal of Pharmaceutical Sciences

Abstract: Background: Bone marrow mesenchymal stem cells (BMSCs) show promise in glioma therapy, but the underlying mechanisms remain unclear. Signal transducer and activator of transcription 3 (STAT3) is a key oncogenic driver in glioma; however, whether BMSCs exert anti-glioma effects through STAT3 modulation requires further investigation. Objectives: To investigate whether BMSCs promote glioma cell apoptosis by inhibiting STAT3 phosphorylation and to validate the causal role of STAT3 using a specific agonist. Methods: C6 glioma-bearing rats were randomized into control, model, BMSC, positive control (MS-275) and STAT3 agonist (Colivelin) groups. BMSCs (1×10⁷ cells) were injected via the tail vein on days 14 and 17. Tumor growth, apoptosis, glial fibrillary acidic protein (GFAP) expression, basic fibroblast growth factor (bFGF)/vascular endothelial growth factor (VEGF) mRNA levels and STAT3/phosphorylated STAT3 (p-STAT3) expression were assessed by magnetic resonance imaging (MRI), histology, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, flow cytometry, immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. In-vitro validation was performed using BMSC-conditioned medium and the STAT3 inhibitor Stattic in C6 cells. Results: BMSC transplantation significantly reduced the tumor area and Ki-67 proliferation index, increased the number of apoptotic cells and the apoptotic rate (P < 0.01) and decreased p-STAT3 levels and the p-STAT3/STAT3 ratio (P < 0.01) without altering total STAT3 expression. Colivelin reversed BMSC-induced apoptosis. bFGF and VEGF mRNA levels were increased in the BMSC group (P < 0.01). In-vitro, BMSC-conditioned medium suppressed STAT3 phosphorylation and induced apoptosis, effects that were enhanced by Stattic. Conclusion: BMSCs promote apoptosis in glioma cells by inhibiting STAT3 phosphorylation. The reversal of these effects by a STAT3 agonist confirms the causal role of STAT3 inhibition. These findings identify STAT3 as a key therapeutic target for BMSC-based glioma therapy.

Abstract: Background: Acute heart failure (AHF) is a leading cause of death from cardiovascular disease and effective treatments are lacking. Objectives: To investigate whether Xinshuaifang improves oxidative stress injury and myocardial remodeling in AHF mice by regulating the gut-heart axis. Methods: C57BL/6 mice aged 8–10 weeks and weighing 20-25 g were randomized into 7 groups (n=8/group): control, AHF model, AHF+Xinshuaifang, AHF+control FMT, AHF+intervention FMT, AHF+Xinshuaifang+antibiotics and antibiotics-only control. AHF was induced by isoproterenol injection (5 mg/kg, 7 days). Gut microbiota composition, oxidative stress markers (MDA, GSH, SOD, MPO), cardiac function (LVDd, LVDs, LVEF, NT-proBNP), myocardial fibrosis and tight junction protein expression (ZO-1, Occludin, Claudin-1, Zonulin) were assessed. Results: Compared with controls, model mice showed significantly reduced Verrucomicrobia and Akkermansia abundance (P<0.01), increased MDA and GSH levels, decreased SOD activity and MPO levels, reduced LVEF, elevated NT-proBNP (P<0.01), decreased ZO-1, Occludin and Claudin-1 expression and increased Zonulin expression. Xinshuaifang intervention significantly reversed all these changes (P<0.01): it increased beneficial bacteria, reduced oxidative stress, improved cardiac function, upregulated ZO-1/Occludin/Claudin-1 and downregulated Zonulin. The FMT intervention group showed similar protective effects, whereas the Xinshuaifang+antibiotics group showed attenuated protection. Conclusion: Xinshuaifang improves cardiac function and alleviates oxidative stress and fibrosis in AHF mice by regulating gut microbiota, upregulating tight junction proteins and restoring intestinal barrier function.

Abstract: Background: Chronic non-atrophic gastritis is a common digestive system disease with a high incidence. While Traditional Chinese Medicine (TCM) offers unique therapeutic approaches, the specific combined efficacy of Woxuan needling and Xiangsha Liujunzi decoction warrants further clinical investigation. Objectives: To explore the effect of Woxuan needling combined with Xiangsha Liujunzi decoction on traditional Chinese medicine syndromes in patients with chronic non-atrophic gastritis. Methods: Convenience sampling was used to select 120 patients with chronic gastritis admitted to Jiangxi Province Hospital of Integrated Chinese and Western Medicine between 1 January 2022 and 1 January 2023. According to the order of enrolment, they were divided into an observation group and a control group using a random number table. The control group received basic drug treatment combined with Xiangsha Liujunzi decoction, whereas the observation group received Woxuan needling in addition to the control treatment. Results: Compared with the control group, the levels of stomach burning, abdominal distension or dull pain, loose stools, fatigue, shortness of breath and poor appetite were significantly lower in the observation group after treatment (p < 0.05). The serum pepsinogen level, gastroscopy score and gastrointestinal symptom rating scale score were also lower, whereas the gastrin level was higher in the observation group (p < 0.05). Conclusion: Treating patients with chronic non-atrophic gastritis with Woxuan needling combined with Xiangsha Liujunzi decoction can improve clinical symptoms and enhance therapeutic efficacy.

Abstract: Background: Tibial transverse osteotomy (TTO) is a promising but limited surgical option for diabetic foot (DF). Berberine, a natural compound from Coptis chinensis, exhibits anti-inflammatory and pro-angiogenic effects relevant to metabolic diseases. Objectives: This study aimed to evaluate whether adjunctive berberine enhances the therapeutic outcomes of TTO in DF patients and to explore its potential association with the platelet-derived growth factor-B (PDGF-B)/PDGF receptor-β (PDGFRβ) signaling pathway. Methods: This was a prospective, single-center, randomized, placebo-controlled, parallel-group, blinded trial. Of 200 screened DF patients, 96 eligible participants (Wagner grade 2–4) scheduled for TTO were randomized 1:1 to receive either TTO plus local berberine injections (n=48) or TTO plus local saline placebo (n=48). An additional non-randomized diabetic reference group (n=10) and healthy control group (n=10) were included for exploratory cellular studies only. The primary outcome was the total effective rate of ulcer healing at 3 months. Secondary outcomes included changes in Ankle-Brachial Index (ABI), ulcer area, Visual Analogue Scale (VAS) pain score and serum inflammatory markers (hs-CRP, IL-6). Exploratory outcomes comprised serum VEGF, bFGF and PDGF-B levels, as well as fibroblast apoptosis in wound tissue. Results: All 96 randomized participants completed the 3-month follow-up, with no dropouts. The berberine group demonstrated a significantly higher total effective rate than the surgery-alone group (87.5% vs. 68.8%; risk difference, 18.7%; 95% CI, 1.2% to 36.2%; P < 0.05). At 3 months, the berberine group also showed greater improvements in ABI (mean difference, 0.13; 95% CI, 0.08 to 0.18; P < 0.01) and ulcer healing rate (mean difference, 16.7%; 95% CI, 9.5% to 23.9%; P < 0.01), along with more significant reductions in hs-CRP and IL-6. Berberine treatment was associated with reduced fibroblast apoptosis in wound tissue (mean difference, -1.10%; 95% CI, -1.56% to -0.64%; P = 0.001) and lower protein expression of PDGF-B and PDGFRβ. No severe adverse events related to berberine injections were reported. Conclusion: In DF patients undergoing TTO, adjunctive local berberine therapy significantly improves the total effective rate and promotes ulcer healing, with benefits associated with reduced inflammation and fibroblast apoptosis and downregulation of the PDGF-B/PDGFRβ pathway. These findings are correlative and require further mechanistic validation. Larger multi-center trials are needed to confirm efficacy and safety.

Abstract: Background: Cardiomyocyte apoptosis plays a critical role in ventricular remodeling and heart failure following acute myocardial infarction (AMI). The Rho kinase signaling pathway is known to be involved in this pathological process. Butylphthalein, a compound with neuroprotective effects, may exert cardioprotective effects by regulating this pathway. Objective: This study aims to investigate the effects of butylphthalein on hemodynamic parameters and cardiomyocyte apoptosis in a rat model of AMI and to explore whether these effects are mediated through the Rho kinase signaling pathway. Methods: Fifty SD rats were randomly divided into five groups: Sham, AMI model, butylphthalein, fasudil (a Rho kinase inhibitor) and combination therapy. AMI was induced by ligation of the left anterior descending coronary artery. Two weeks post-surgery, hemodynamic parameters—including left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP) and maximal rate of rise and fall of ventricular pressure (±dp/dtmax)—were measured. Cardiomyocyte apoptosis was assessed using TUNEL staining. The expression levels of Bcl-2, Bax, RhoA, ROCK1 and the p-MYPT1/MYPT1 ratio were detected by Western blot. Results: Rats in the AMI model group exhibited significant hemodynamic impairment and increased cardiomyocyte apoptosis. Compared with the model group, butylphthalein treatment significantly improved LVSP and ±dp/dtmax, reduced LVEDP, decreased the number of TUNEL-positive cells, upregulated Bcl-2 expression, downregulated Bax expression and suppressed the expression of RhoA/ROCK1 and the p-MYPT1/MYPT1 ratio (all P<0.05). The combination of butylphthalein and fasudil resulted in enhanced therapeutic effects, suggesting a synergistic inhibition of Rho kinase activity. Conclusion: Butylphthalein attenuates cardiomyocyte apoptosis and improves cardiac function in rats with AMI, likely through inhibition of the Rho kinase signaling pathway. These findings support its potential as a therapeutic agent for AMI.All experiments were performed on male Sprague-Dawley rats (n=50).