Ethnopharmacological and Cancer Chemoprevention Investigation of Selected Flora of Southern Punjab, Pakistan

Ethnomedicinal studies are significant for the discovery of new drugs from indigenous medicinal plants. The current study aimed to report the indigenous knowledge regarding medicinal plants and herbal remedies used as folk medicines in Cholistan desert, Vehari, Layyah, Bahawalnagar and Rajanpur districts of Punjab Province, Pakistan. Further the cancer chemoprevention potential of the selected flora have been evaluated. Rapid Appraisal Approach (RAA), semi-structured interviews, group meetings with herbalists, landowners and local people about the medicinal plants were employed to collect the data. Plants were dried under shade, ground into fine powder and soaked for 7 days at room temperature in organic solvents filtered and the filtrates were dried under reduced pressure at 45°C in rotary evaporator. The residue called "crude extracts" were used for chemopreventive assays such as Inhibition of TNF-? activated nuclear factor-kappa B (NFkB), aromatase inhibition, Inhibition of nitric oxide production (iNOp), Induction of Quinone reductase 1 (QR1), DPPH free radical scavenging activity and cytotoxicity against Hepa 1c1c7, two neuroblastoma cell lines MYCN-2, SK-N-SH and two breast cancer cell cultures MCF-7 & MDA-MB-231. 400 medicinal plants belonging to 179 families disseminated among 349 genera were reported in the study. Poaceae was the predominant family with 13 reported medicinal plants in Cholistan desert. Leaves were the dominant part of the plant with 26.4% usage in herbal preparations followed by the stem (25.2%), fruit (21.5%), flower (16.3%), seed (6.5%), bark and pod (02%). Haloxylon recurvum exhibited the highest used vale (UV) 0.83 while least UV (0.16) was exhibited by Mollugo cerviana. Poaceae was the predominant family with 16 reported medicinal plants in Layyah district. Wild herbaceous plants were predominant and the leaves contributed 32% followed by the stem (27%), fruit and flowers (15%). Allium sativum exhibited the highest use value (UV) 0.92 while lowest UV (0.15) was exhibited by Lasiurus scindicus. Fabaceae is the predominant family with 9 reported medicinal plant species in Vehari district. The maximum use of leaves recorded was 30.12% followed by stem 24.62%, fruit 14.22%, flower 12.97%, seeds 12.13%, bark 4.6% and pod 1.25%, in herbal preparations. Allium cepa exhibited the highest use value (0.90) while lowest UV (0.10) was presented by Aerva javanica. Fabaceae was the predominant family contributing 09 plants species effective for the treatments in the local healthcare system of Bhawalnagar district. A total of 65 medicinal plants belonging to 56 genera and 34 families were reported to be effective for 08 major ailments categories in the local healthcare system of Rajanpur district, Punjab Province, Pakistan. Acacia arabica exhibited highest used value while Zizyphus spina showed high Cultural Index (CI), Relative Index (RI) and Cultural Value Index (CVI). Informant Census Factor (ICF) for respiratory system ailments category was 0.80. The aerial parts of the selected medicinal plants from study area have ability to suppress the factors TNF-?-NFkB, Aromatase inhibition, inhibition of nitric oxide production (iNOp), scavenge free radicals (DPPH), Quinone Reduction (QR1) and posses the anticancer activity against Hepa 1c1c7 cell line, two neuroblastoma cell lines MYCN-2, SK-N-SH and two breast cancer cell lines MCF-7 and MDA-MB-231 used. The crude extracts of Calotropis procera, Fagonia indica and Tamarix aphylla suppressed TNF-?-NFkB, extracts of Fagonia indica, Suaeda fruiticosa, Capparis deciduas, Euphorbia heliscopia, Salvadora oleoides and Calotropis procera showed promising inhibition of aromatase and crude extracts of Fagonia indica and Capparis deciduas exhibited inhibition of nitric oxide production while extracts of Capparis deciduas, Solanum surratense, Euphorbia heliscopia, Tamarix aphylla, Fagonia indica, Salsola baryosma, Salvadora oleoides and Aerva javanica showed significant induction of Quinone reductase enzyme 1 at 20 µg/mL. Extracts of Salsola baryosma, Calotropis procera and Tamarix aphylla exhibited significant DPPH free radical scavenging activity at 200 µg/ml. n-Hexane, Acetone, Ethanol and Ethanol: Water extracts of Fagonia indica, Suaeda fruiticosa, Capparis deciduas, Euphorbia heliscopia, Salvadora oleoides and Calotropis procera and Tamarix aphylla are considered to be best potent chemopreventive agents. The selected plants also showed cytotoxic potentials to Hepa 1c1c7, MYCN-2, SK-N-SH, MCF-7 and MDA-MB-231 in cell cultures at 20µg/mL. The identification of bioactive constituents of these selected plant extracts is needed for pharmaceutical applications in drug delivery and identification of bioactive metabolites of selected flora of Cholistan desert is already in progress