Background Cynodon dactylon (Poaceae), a perennial herb used in traditional Indian medicine, is widely used to treat infections, inflammatory disorders, skin diseases, and wound-related ailments. Despite its ethnomedicinal use, systematic scientific evidence linking its phytochemical composition to multitarget pharmacological activity remains limited. This study aimed to evaluate the pharmacological potential and chemical basis of ethanol and hexane extracts from the whole C. dactylon plant. Methods Whole plants of C. dactylon collected from Kerala, India, were authenticated and extracted using Soxhlet extraction with ethanol and hexane. Quantitative phytochemical assays were performed to determine the total phenolic, flavonoid, tannin, and alkaloid contents. Wound-healing activity was evaluated in L929 fibroblast cells using an in vitro scratch assay. Antimicrobial activity was assessed using two-fold microdilution method against Staphylococcus aureus, Escherichia coli, and Candida albicans. Anti-inflammatory activity was examined by measuring nitric oxide (NO) inhibition in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Antioxidant activity was determined using DPPH radical-scavenging assay. Anthelmintic activity was tested against Pheretima posthuma, and the biocompatibility was evaluated in L929 cells using the MTT assay. Phytochemical profiling of the ethanol extract was performed using LC–MS/MS analysis. Results Quantitative phytochemical analysis revealed solvent-dependent variations, with higher phenolic (90.82 ± 2.14 µg GAE/mg) and tannin (99.10 ± 1.87 µg TAE/mg) contents in CdE, while alkaloids were higher in CdH (94.5 ± 2.06 µg AE/mg). In the in vitro scratch assay, CdE promoted complete wound closure at 100 µg/mL within 36 h, whereas CdH achieved 68% wound closure (p < 0.05). Antimicrobial assays revealed that CdE exhibited superior efficacy, with MIC₅₀ values of 118 ± 2.1, 236 ± 1.7, and 303 ± 2.0 µg/mL against Staphylococcus aureus, Escherichia coli, and Candida albicans, respectively. In contrast, CdH demonstrated MIC₅₀ values of 732 ± 2.0, 541 ± 2.6, and 318 ± 2.1 µg/mL, indicating comparatively weaker activities. CdE also inhibited nitric oxide production in LPS-stimulated RAW 264.7 macrophages in a concentration-dependent manner. Moderate antioxidant activity was observed for CdE (IC₅₀ = 890.42 µg/mL). Both extracts exhibited anthelmintic activity against Pheretima posthuma and demonstrated high biocompatibility (> 95% cell viability). LC–MS/MS profiling of CdE identified 18 polyphenolic compounds. The observed pharmacological activities were mechanistically interconnected, with antimicrobial, anti-inflammatory, and antioxidant effects collectively enhancing the wound-healing response. The identified polyphenolic constituents provide a chemical basis for these activities, with compounds such as shikimic acid, cinnamic acid, and ferulic acid being suggested as potential lead molecules. These findings underscore the multi-target mode of action mediated by the synergistic interaction of bioactive phytoconstituents. Conclusion The results indicate that CdE exhibits notable wound-healing, antimicrobial, anti-inflammatory, antioxidant, and anthelmintic activities, with high biocompatibility in in vitro models. These biological effects may be associated with the presence of polyphenolic compounds in the extracts. Overall, our findings support the pharmacological potential of C. dactylon and provide a basis for further studies to elucidate the underlying mechanisms and evaluate its potential for natural product-based therapeutic development.

Phytochemical analysis and potential therapeutic applications of Cynodon dactylon extracts: a multifaceted in vitro pharmacological investigation

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发表日期：

2026年6月25日

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Sheeba Baby Alex,
M. S. Latha

摘要

Background

Cynodon dactylon (Poaceae), a perennial herb used in traditional Indian medicine, is widely used to treat infections, inflammatory disorders, skin diseases, and wound-related ailments. Despite its ethnomedicinal use, systematic scientific evidence linking its phytochemical composition to multitarget pharmacological activity remains limited. This study aimed to evaluate the pharmacological potential and chemical basis of ethanol and hexane extracts from the whole C. dactylon plant.

Methods

Whole plants of C. dactylon collected from Kerala, India, were authenticated and extracted using Soxhlet extraction with ethanol and hexane. Quantitative phytochemical assays were performed to determine the total phenolic, flavonoid, tannin, and alkaloid contents. Wound-healing activity was evaluated in L929 fibroblast cells using an in vitro scratch assay. Antimicrobial activity was assessed using two-fold microdilution method against Staphylococcus aureus, Escherichia coli, and Candida albicans. Anti-inflammatory activity was examined by measuring nitric oxide (NO) inhibition in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Antioxidant activity was determined using DPPH radical-scavenging assay. Anthelmintic activity was tested against Pheretima posthuma, and the biocompatibility was evaluated in L929 cells using the MTT assay. Phytochemical profiling of the ethanol extract was performed using LC–MS/MS analysis.

Results

Quantitative phytochemical analysis revealed solvent-dependent variations, with higher phenolic (90.82 ± 2.14 µg GAE/mg) and tannin (99.10 ± 1.87 µg TAE/mg) contents in CdE, while alkaloids were higher in CdH (94.5 ± 2.06 µg AE/mg). In the in vitro scratch assay, CdE promoted complete wound closure at 100 µg/mL within 36 h, whereas CdH achieved 68% wound closure (p < 0.05). Antimicrobial assays revealed that CdE exhibited superior efficacy, with MIC₅₀ values of 118 ± 2.1, 236 ± 1.7, and 303 ± 2.0 µg/mL against Staphylococcus aureus, Escherichia coli, and Candida albicans, respectively. In contrast, CdH demonstrated MIC₅₀ values of 732 ± 2.0, 541 ± 2.6, and 318 ± 2.1 µg/mL, indicating comparatively weaker activities. CdE also inhibited nitric oxide production in LPS-stimulated RAW 264.7 macrophages in a concentration-dependent manner. Moderate antioxidant activity was observed for CdE (IC₅₀ = 890.42 µg/mL). Both extracts exhibited anthelmintic activity against Pheretima posthuma and demonstrated high biocompatibility (> 95% cell viability). LC–MS/MS profiling of CdE identified 18 polyphenolic compounds. The observed pharmacological activities were mechanistically interconnected, with antimicrobial, anti-inflammatory, and antioxidant effects collectively enhancing the wound-healing response. The identified polyphenolic constituents provide a chemical basis for these activities, with compounds such as shikimic acid, cinnamic acid, and ferulic acid being suggested as potential lead molecules. These findings underscore the multi-target mode of action mediated by the synergistic interaction of bioactive phytoconstituents.

Conclusion

The results indicate that CdE exhibits notable wound-healing, antimicrobial, anti-inflammatory, antioxidant, and anthelmintic activities, with high biocompatibility in in vitro models. These biological effects may be associated with the presence of polyphenolic compounds in the extracts. Overall, our findings support the pharmacological potential of C. dactylon and provide a basis for further studies to elucidate the underlying mechanisms and evaluate its potential for natural product-based therapeutic development.