Anticancer immune responses mediated through bioactive phytochemicals from Datura stramonium of Western Himalayas an insight into immune cells

Abstract

newline xi newlineABSTRACT newlineDatura stramonium L. is a miraculous Ayurvedic herb used in the treatment of innumerable diseases. Various studies have revealed the anti-inflammatory potential of this plant but not much work has been done on the chemical constituents which are responsible for its anti-inflammatory effects. Therefore, the current study deals with the identification and isolation of the D. stramonium lactones and essential oil from the leaves. The compounds were isolated from D. stramonium leaves and then characterized by NMR, HR-ESIMS and HPLC techniques. Three Datura lactones were isolated and identified as daturalactone (D1), 12 deoxywithastramonolide (D23) and daturilin (D27). The GC-MS analysis showed that the dominant compounds present in the D. oil were neophytadiene (phytol acetate) (10.76 %), damascenone (9.67 %), eudesmol (7.2 %). The antioxidant activity of lactones (D1, D23 and D27 at 25-200 and#956;M) and D. oil (10 to 80 and#956;g/ml) was analyzed by ABTS and DPPH assay. Our results show that the lactones were able to scavenge less free radicals while D. oil had a promising free radicals scavenging as measured by DPPH (IC50 71.35 ±1.06 and#956;g/ml) and ABTS (IC50 61.01 ± 1.07 and#956;g/ml) assays. newlineFurther, the compounds were evaluated for their anti-inflammatory activity using in vitro/in vivo/in silico model. In Bovine serum albumin (BSA) inhibition and Red Blood Cell (RBC) membrane stability assays, the three lactones and D. oil were capable of inhibiting the protein denaturation and RBC lysis. In vitro assay showed that these lactones (10 and#956;M) and D. oil (10 and#956;g) significantly inhibited the LPS induced inflammation in J774A.1 macrophage cells via suppressing the levels of pro-inflammatory cytokines tumor necrosis factor-and#945; (TNF-and#945;), interleukin-1and#946; (IL-1and#946;), interleukin-6 (IL-6) and nitric oxide (NO) with a relative activity of D1gtD23gt D27. Moreover, in vivo assays depicted that D1, D23 and D27 (20 mg/Kg) were able to reduce the pain and inflammation in Tail flick, acetic acid induced writhing, Carrageenan-induced hind