Immunomodulatory activity and phytochemical analysis of selected plants used in folk medicine of palakkad for post natal care

Abstract

The study investigates the immunomodulatory and phytochemical profiling of selected plants used in the folk medicine of Palakkad for postnatal care. The traditional medicinal practices play a vital role in postnatal care of Kerala. Our study mainly focussed on plants used for making vethuvellam, a traditional medicated water for bathing mothers after delivery. The study documented the plants used for bathing mothers and infants after delivery. The medicated bath enhances immunity, reduces inflammations and prevents infections. Despite the traditional significance, scientific validation is lacking in this aspect. Our study aimed to document the plants used for traditional bathing of mother and child in Palakkad district, evaluate the bioactivities of selected plants and analyze the phytochemical profile of selected plants. A survey was conducted on plants used for bathing mother and child in various parts of Palakkad district. 282 informants from the district participated in the survey. 62 plant species used for bathing mother and child after delivery were documented from the survey. The study also reported the various combinations of plants used for making vethuvellam. Quantitative ethnobotanical tools like Relative frequency of citation (RFC), Consensus index (CI) and Use value (UV) were used to express the collected data. Curcuma longa and Artocarpus heterophyllus showed high RFC, CI and UV. Acalypha fruticosa Forssk., Justicia gendarussa Burm. f., Piper longum L. and Vitex altissima L. f. were selected for further studies based on the survey and prior literature. Leaf extracts of these selected plants in hexane, water and isopropyl alcohol were used in the study. The preliminary phytochemical analysis was done using standard qualitative test. Preliminary phytochemical screening confirmed the presence of alkaloids, flavonoids, tannins, phenolics and saponins across the extracts. This research investigated the immunomodulation properties of 4 selected plants using in vitro macrophage activation studies in THP1 cells. All the four plants modulate the immune system via upregulating or down regulating the expression of IL-1β, IFNβ, TNFα, and CCR-5. V. altissima was selected for detailed immunomodulation studies. The hexane (VAH) and isopropyl alcohol (VAI) extract of V. altissima upregulated the gene expression of IL-1β, IFNβ, TNFα, and CCR-5 indicating the immunostimulatory property of both extracts. Our research also studied the antioxidant and antibacterial properties of three different extracts of V. altissima. DPPH assay was used to study the antioxidant activity Our research also studied the antioxidant and antibacterial properties of three different extracts of V. altissima. DPPH assay was used to study the antioxidant activity and agar disc diffusion studied against Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus was used to explore the antibacterial activity. The V. altissima isopropyl alcohol (VAI) extract exhibited superior antioxidant and antibacterial activities. This study also analysed Orbitrap high resolution liquid chromatography mass spectrometry (OHR-LCMS) for compound identification from V. altissima hexane, water (VAW) and isopropyl alcohol extracts. 89 compounds were obtained from VAW, 55 compounds were obtained from VAH and 55 compounds were obtained from VAI. Majority of these compounds possessed many bioactivities. Our study also highlighted the anti-inflammatory potential of V. altissima isopropyl alcohol extract using lipopolysaccharide (LPS) stimulated THP1 derived macrophages. The extract significantly downregulated the expression of IL-1β, IFNβ, TNFα, IL-6 and CCR-5 compared to LPS stimulated control group. The V. altissima isopropyl alcohol extract also exhibited antifungal activity against Aspergillus niger studied using food poison technique. The TLC analysis of VAI extract revealed the presence of seven compounds. The antibacterial activity of TLC fractions was also studied against B. subtilis, E. coli, P. aeruginosa and S. aureus. The fractions C5, C6, and C7 showed maximum antibacterial activity against E. coli and B. subtilis. This study validates the importance of vethuvellam in postnatal care of Palakkad district. V. altissima emerges as a leading candidate for drug development due to its immunostimulatory, anti-inflammatory, antibacterial, antioxidant and antifungal activities. The research also supports the folk medicinal use of this plant. The documentation of folk medicinal plants for postnatal care is essential to protect the traditional knowledge.