Analysis of Diosgenin Content and Pharmacological Exploration of Endangered Medicinal Herb Trillium Govanianum Nag Chhatri

Abstract

Trillium govanianum Wall. ex. D. Don, is a high value medicinal herb of North-Western Himalayan regions of India having plethora of medicinal properties like Cancer, Sexual disorders, GI disorders, skin infections, Anthelmintic, etc. The rhizome part of the plant containing trillarin is of commercial value to pharmaceutical industry. Diosgenin which is a phytoestrogen can be chemically converted into antifertility drugs having their steroidal base such as progesterone, corticosteroids and anabolic steroids. This species has high folkloric value and rhizome of this plant species are used for treating wounds, dysentery, skin boils, infections, menstrual and sexual disorders. Overexploitationofthesespeciesnecessitatesthedevelopmentofconservationstrategiesandenhancedproductionofdiosgenin at commercial scale. newlineSince limited studies have been carried on T. govanianum rhizome and soxhlet extraction and optimization, we studied in detail and devised a novel methodology for the same. The detailed study was carried on getting the highest extractive yield of 43.56% (30:70; Water: Methanol) using different solvent systems depending upon eluotropic series w.r.to maximum diosgenin content (2.4 %). Finally by using response surface methodology (RSM) best condition viz. incubation time for hydrolysis (8 hr), temperature (850C) and solid- liquid ratio (234.09 gm/ml) were optimized giving maximum diosgenin content (5.99 %). newlineMolecularbasisofdiosgenin biosynthesis was exploredinT. govanianumfor which, partial gene sequences of 5 genes encoding key enzymes viz. HMG Co-A reductase (HMGR), squalene synthase (SQS), farnesyl pyrophosphate synthase (FPPS), 26-O-beta-glucosidase (BETA) and cycloartenol synthase (CAS), involved in diosgenin biosynthesis pathway were cloned through comparative genomics. The RT-qPCR analysis revealed that all five genes viz. HMGR, FPPS, SQS, BETA and CAS involved in diosgenin biosynthesis had relatively higher expression in the rhizomes of Chamba (2.4%) (up to 3.2 folds) (plt0.05) as compared to Lahaul and Sp