Pakistan Journal of Pharmaceutical Sciences

Abstract: Background: Dental caries are chronic infectious diseases with a high global prevalence. Its pathogenesis is closely related to the adhesion of Streptococcus mutans (S. mutans), formation of dental plaque biofilms and acid production. Currently, only a few types of anticaries drugs are available for clinical use and the long-term use of these drugs can lead to side effects, such as drug resistance and dysbiosis of the oral microbiota. Objectives: Therefore, exploring new, safe and effective anti-caries treatments is an urgent task. Screening antimicrobial drugs from herbal medicines for non-antibiotic treatment of bacteria could provide new solutions to these problems. This study aimed to screen food-borne herbal extracts with synergistic antimicrobial potential to provide a theoretical foundation for the development of natural anticaries drugs. Methods: This in-vitro study evaluated the minimum inhibitory concentration and minimum biofilm inhibitory concentration of individual plant extracts (cinnamaldehyde, eugenol and polydatin) and a 1:1 mixture of these extracts. The effects of drug combinations were assessed using the checkerboard method. The inhibitory effects of drugs against S. mutans and their antibacterial mechanisms were evaluated through scanning electron microscopy and conductivity measurements. Their ability to inhibit S. mutans biofilms was assessed through crystal violet (Beijing Solebao Technology Co.) staining and scanning electron microscopy. Their inhibition of bacterial acid production and cytotoxicity to human gingival fibroblasts were also examined. Results: The combination groups exhibited synergistic bacteriostatic effects while reducing cytotoxicity. Individual and combined plant extracts exerted bacteriostatic effects by disrupting the bacterial cell membrane, significantly inhibiting S. mutans biofilm formation while suppressing its acid production. Conclusions: The application of combined food-borne herbal extracts inhibits the growth of S. mutans and its biofilm synergistically while reducing the required drug concentration and cytotoxicity. This provides a new approach for the development of natural anticaries preparations.

Abstract: Background: Fibrotic liver is one of the serious pathophysiological consequences of chronic injury to the liver, may progress toward serious complications including cirrhosis and hepatic carcinoma. Objectives: Panax notoginseng is used for both therapeutic and prophylactic purposes in TCM; it is a source of Dencichine, an active substance that has been proven to exhibit antifibrotic properties in hepatic fibrosis. The present study investigated how Dencichine treatment ameliorates liver fibrosis using an animal model and a cellular fibrosis model. Methods: Randomly assigned, male C57BL/6J mice received 10% CCl4 for 15 consecutive weeks to induce hepatic fibrosis. Treatment with Dencichine was initiated from 7th week onward. To simulate in-vitro the presence of a fibrotic microenvironment, LX-2 cells were treated with PDGF-BB at a concentration of 20 ng/mL and incubated from one up to four days and then treated by various doses of Dencichine. Liver function was evaluated by measuring alanine aminotransferase (ALT), AST, HA and LN. Histopathology was evaluated by haematoxylin/eosin (HE), Masson’s trichrome and Sirius red stains. Quantitative real-time PCR and western blot analyses were used to evaluate mRNA and protein expression levels, respectively. Wound-healing assays and transwell chamber experiments were used to assess cell migration; EdU proliferation detection and flow cytometry were performed to measure cell proliferation and cell cycle, respectively. Results: The most important results are a considerable reduction of serum concentrations of AST and ALT, HA and laminin in CCl4-treated mice decreased liver injury and ECM deposition. Furthermore, this compound downregulated the expression of cyclin D1 and cyclin E1 and blocked the activation of PI3K and AKT kinases in fibrotic liver cells. Conclusion: These results indicate that Dencichine may slow the development of liver fibrosis by interfering with the PI3K-AKT signaling cascade, demonstrating significant anti-scarring properties.

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.