Tinospora (PROSEA)

Plant Resources of South-East Asia
Introduction

List of species

Tinospora Miers

Protologue: Ann. Mag. Nat. Hist. ser. 2, 7: 35 (1851).

Family: Menispermaceae

Chromosome number: x= unknown; T. cordifolia: 2n= 24, 26

Major species

Tinospora crispa (L.) Hook.f. & Thomson,
T. glabra (Burm.f.) Merr.

Vernacular names

Philippines: makabuhay (general).

Origin and geographic distribution

Tinospora consists of 33 species, which occur in the tropical and subtropical parts of the Old World: 7 species in tropical Africa, 2 in Madagascar, and 24 in Asia extending to Australia and the Pacific. In Malesia, 14 species have been found, but most of them are known from few collections; only T. crispa and T. glabra are widespread. T. crispa is also cultivated as a medicinal plant, e.g. in Thailand, Sri Lanka and India.

Uses

Throughout most of South-East Asia, Tinospora species are a widely acclaimed source of remedies for many different complaints. Much of their reputation is probably due to influences from Chinese traditional medicine, where for instance stems and leaves of T. crispa are valued for their anti-inflammatory (antirheumatic), febrifugal, antimalarial and antibacterial properties. In Indonesia, Malaysia, Thailand and the Philippines, stems (infusions, decoctions) of T. crispa are considered to be effective in the treatment of skin complaints (external as antiseptic, antiparasitic, and for treating wounds and itches), stomach complaints (ulcers, as appetizer, tonic), diarrhoea, fevers (e.g. malaria, smallpox), diabetes mellitus, cholera (whole plant), jaundice, and as a vermifuge (also in children). In veterinary medicine, T. crispa is applied as veterinary tonic; powdered stems are used to fatten horses and cattle by stimulating their appetite.

T. glabra has similar uses as T. crispa although the latter is thought to be more effective, but is recommended particularly for dealing with wounds, scabies and tropical ulcers.

It is perhaps in the Indian subcontinent where the widest use is made of Tinospora. The main species is T. cordifolia (Willd.) Miers, which does not occur in Malesia. In present-day Ayurvedic medicine in India, all parts of the plant find wide use for their general tonic, anti-inflammatory, anti-arthritic, anti-allergic, antimalarial, antidiabetic, hepato-protective and aphrodisiac properties. Categorized as "rasayana" in Ayurveda, it is also used for its general adaptogenic and pro-host immuno-modulatory activity in fighting infections. T. cordifolia is thus claimed to be useful in e.g. skin diseases, jaundice, diabetes, anaemia, fever and rheumatism. In fact the plant is part of almost all decoctions mentioned in Ayurvedic textbooks for use in joint diseases. The starch from the stems and roots is used as a nutrient in chronic diarrhoea and dysentery. Juice of the fresh plant is a powerful diuretic (urinary diseases), and also used in gonorrhoea with advantage. Besides its antimalarial activity, the root is known for its antistress and antileprotic activities. Finally, T. cordifolia is also used as veterinary medicine.

A tincture from the stem of T. sinensis (Lour.) Merr. (synonyms: T. malabarica (Lamk.) Hook.f. & Thomson, T. tomentosa (Colebr.) Hook.f. & Thomson) has considerable reputation in Indo-China for treating arthritis and chronic rheumatism. There are also reports suggesting the efficacy of the roots, stems and leaves in conditions like cough, wound healing, malaria, skin complaints and allergic disorders.

Production and international trade

Stem parts or powdered stems can be found frequently on local markets, but are not traded internationally.

Properties

A number of chemical constituents have already been isolated from different parts of T. crispa, e.g. diterpenes, alkaloids and flavonoids. Most extensively investigated are a series of furano-diterpene (glycoside) compounds: tinotufolin A-F (leaves), borapetoside A-H (glycosides, stems), borapetol A, B (stems, aglycones of borapetoside A, B) and the bitter tinocrisposide (glycoside, stems). In addition to these furano-diterpenes, a series of clerodane-diterpene glycosides has also been isolated from the stems and named rumphioside A, B, C, C-1, Ac-D, E and F.

Amongst the alkaloids isolated from T. crispa are the well known protoberberine type alkaloids palmatine (stems), berberine (stems, aerial roots) and the aporphine type alkaloid tembetarine (stems, aerial roots). From the methanolic stem extract three further N-acyl-aporphine type alkaloids have been isolated and their structures elucidated: (-)-N-formyl-annonaine, (-)-N-formyl-nornuciferine (= tinocrispicine) and (-)-N-acetyl-nornuciferine. Of these isolated alkaloids, berberine and its salts are known to have spasmolytic, antibacterial and in some degree antifungal and antiprotozoal activity.

Further constituents from T. crispa include: N-trans and N-cis feruloyl tyramine (stems, phenolic acid amides), tinotuberide (stems, phenolic glucoside), and the flavone-O-glycosides (from stems): luteolin-4'-methylether-7-glucoside, genkwanin-7-glucoside, luteoline-4'-methylether-3'-glucoside, diosmetin and genkwanin.

An aqueous extract from T. crispa stems showed lowering of blood glucose levels and stimulated insulin release in moderate alloxan-diabetic rats. The dose administered orally (4 g/l in drinking water) and the method of preparation of the extract were comparable to those used by diabetics in Malaysia. After two weeks of treatment, the rats also showed an improvement in the glucose tolerance test; there were no effects in normal and severe alloxan-diabetic rats. These results suggest that the hypoglycaemic effect observed is due to stimulation of insulin release, rather than some extra-pancreatic action. This insulinotropic activity was also observed after intravenous injection of 50 mg/kg of the extract in normal rats.

Toxicity of Tinospora extracts proved to be low in tests with rats (LD₅₀ values of over 5 g/kg orally, 3 g/kg dermally). Tinospora extracts in high concentrations can cause infection in the liver and follicular atresia in mice, but the kidneys are not much affected. Crude extracts showed protective effects in mice with experimental urinary tract infection caused by Pseudomonas aeruginosa.

A number of compounds have been isolated from the stems of T. cordifolia, including diterpenes and alkaloids. The diterpenes comprise the major group of components isolated: the norditerpene-furan-glycosides cordifoliside A-E, the clerodane-diterpenes tinosponone, tinosporaside (= tinosponone-glycoside) and tinocordioside (glycoside) and the clerodane-furano-diterpenes cordioside (glycoside), together with 4 other furano-diterpene compounds: colombin, one being an epimer of 6-hydroxy-arcangelisin, and two being a set of optical isomers (not yet named). The biosynthesis of the clerodane-furano-diterpene skeleton has also been investigated.

Amongst the alkaloids isolated from T. cordifolia are magnoflorine and tembetarine (stems, aporphine type alkaloids) and jatrorrhizine (roots, protoberberine type alkaloid). Syringin is a phenolic component, isolated from the fresh stems.

Considerable efforts have been made to investigate the biological effects of T. cordifolia. An aqueous, alcoholic and chloroform extract of the leaves after oral application exerted a significant hypoglycaemic effect in both normal and alloxan-diabetic rabbits. The reaction in alloxan-diabetic animals, in which almost all pancreatic β-cells have been destroyed, appears to be like a direct effect, probably by a mechanism similar to insulin. However, the stronger effect in normo-glycaemic rabbits suggests that the mechanism of action consists not only of this direct insulin-like effect, but that there is also an indirect action by stimulating the insulin release from pancreatic β-cells. Besides the hypoglycaemic activity, no effects were found on the blood lipid levels, and acute toxicity studies did not reveal visible signs and symptoms of toxicity. Further investigations in albino rats and different groups of rabbits confirm the hypoglycaemic effect and the proposed mechanism of action.

Several studies have focused on the immunotherapeutic effects. Pretreatment (oral) with an aqueous extract strongly reduced the mortality in a mouse model for E. coli peritonitis. This was associated with significantly improved bacterial clearance and improved phagocytic capacities of neutrophils in the group treated with T. cordifolia. T. cordifolia extract itself did not possess in vitro bactericidal activity. These results were confirmed in experiments with wistar rats in which abdominal sepsis was induced by caecal ligation. Both in rat and man, oral application of a T. cordifolia extract improved the surgical outcome in patients with obstructive jaundice, in which sepsis initiated by bacteria in the bile at the time of biliary tract surgery comprises a major risk. In the group receiving the extract, the phagocytic and killing capacities of neutrophils improved; thus T. cordifolia appears to act by strengthening host defences.

Further investigations included antileishmanial, anti-stress, antipyretic and diuretic effects. Leishmaniasis, commonly known as "kala-azar", causes great mortality in tropical and subtropical regions of the world. The antileishmanial effect was evaluated in infected golden hamsters. A 50% ethanol extract of T. cordifolia stems showed significant inhibition of multiplication of parasites, and increased survival periods. The anti-stress activity was studied by investigating the effects of treatment on brain norepinephrine, dopamine and serotonine levels in stressed rats. The ethanol extract, prepared from roots previously extracted with petrol (60-80) and chloroform, and given orally after drying, was found to possess normalizing activity against stress-induced changes on brain neurotransmitter levels. The antipyretic effect was investigated in rats, and yeast-induced pyrexia was used to screen the extracts. The antipyretic action was clearly significant for a T. cordifolia ethanol extract (whole plant, given orally) and appeared to be comparable to that of the aspirin control. Finally, T. cordifolia extract showed mild diuresis and a significant increase in the excretion of electrolytes in rats after oral application.

Tinospora extracts are toxic to brown planthoppers (Nilaparvata lugens) and green leafhoppers (Nephotettis virescens), which are common pests in rice in the Philippines. Soaking the roots of rice seedlings in aqueous Tinospora extract in the field is effective in controlling the major rice pests, and broadcasting the ground stems on the seed-beds 10 days after sowing is also applied successfully. The extracts can control the diamondback moth (Plutella xylostella); the effectivity is comparable to malathion, and the extracts show antifeedant effects on the insect. In tests in Thailand, T. crispa extract showed mild repellency to the oviposition of the oriental fruit fly (Darcus dorsalis). It is also effective in controlling borers on maize, and bollworms (Helicoverpa armigera) in cotton. The extract showed high toxicity to fish in experiments in Malaysia. In vitro tests in the Philippines using mutant strains of Bacillus subtilis and Salmonella typhimurium showed that Tinospora extracts do not contain direct mutagens.

Adulterations and substitutes

Other Menispermaceae (e.g. Cyclea and Stephania species) have similar or related alkaloids and have similar applications. Other insecticides of plant origin used in South-East Asia are present in the roots of Derris and Lonchocarpus spp., in seeds of Croton tiglium L., in leaves of Azadirachta indica A.H.L. Juss., Nicotiana tabacum L. and Vitex negundo L., in leaves, fruits and bark of Melia azedarach L., in whole plants of Tanacetum cinerariifolium (Trev.) Schultz-Bip., Lantana spp. and Tagetes spp., and in leaves, roots and seeds of Annona squamosa L.

Description

Dioecious woody climbers up to 15 m long, sometimes scandent shrubs, usually entirely glabrous; stem woody, with bark often becoming detached on drying, striate when young, becoming tuberculate or warty with raised lenticels, usually glabrous but sometimes puberulous, sometimes producing very long filiform aerial roots.
Leaves arranged spirally, simple and entire (occasionally dentate or 3-lobed), often cordate, palmately veined, sometimes domatia present in axils of veins beneath, petiole swollen and geniculate at base; stipules absent.
Inflorescence axillary or cauliflorous, thyrsoid, pseudopaniculate, pseudoracemose or pseudospicate.
Flowers unisexual, 6-merous; sepals usually free, outer 3 usually smaller, elliptical, imbricate; petals free, often broadly cuneate-ovate with the lateral edges inrolled, usually fleshy and often glandular-papillose externally towards the base; male flowers with 6 free stamens; female flowers with 3 curved-ellipsoid carpels having short-lobed, reflexed stigmas, and 6 subulate staminodes.
Fruit a usually ellipsoidal drupe with terminal style scar, borne on a short or columnar carpophore; endocarp bony, dorsally convex and often verrucose or tuberculate, ventrally with central aperture or with shallow longitudinal groove.
Seed with usually ruminate endosperm.

Growth and development

T. crispa usually flowers when leafless, in Thailand in January to March. The scented flowers are pollinated by insects such as small flies and bees, and possibly also small beetles and moths. Female plants, with or without fruits, have rarely been collected.

Other botanical information

Tinospora belongs to the tribe Tinosporeae, characterized by the drupe with terminal style scar, and embryo with foliaceous cotyledons, together with Chlaenandra, Fibraurea, Parabaena and Tinomiscium.

T. glabra has been much confused with T. crispa. Information on medicinal and chemical properties of T. crispa (or its synonym T. rumphii) given in literature for the Philippines partly refers to T. glabra.

Ecology

Tinospora occurs in forest, thickets and hedges up to 1000 m altitude. It has a very strong capacity of regeneration and is often very abundant in secondary regrowth after disturbance of the natural forest. In primary forest it occurs much more scattered. T. glabra can also be common in littoral forest and on limestone.

Propagation and planting

Stems of T. crispa remain viable when cut into pieces because the dried sap effectively seals the cut ends. If kept in a closed box they can still be viable after one year. Apparently T. crispa is mainly propagated vegetatively.

In vitro production of active compounds

Experiments on the in vitro culture of Tinospora root segments showed promising results, with good callus formation and a high content of therapeutic substances.

Diseases and pests

Larvae of the noctuid moth Othreis fullonia feed mainly on Tinospora leaves. The adult moth can cause considerable damage to commercial fruit tree plantations, especially of longan (Dimocarpus longan Lour.) and citrus in Thailand, by piercing the skin of the fruits. The destruction of natural forest and the increasing area of secondary vegetation, preferred by Tinospora, promote the spread of the pest.

In Pakistan, T. cordifolia showed effective resistance to the root-knot nematodes Meloidogyne incognita and M. javanica.

Handling after harvest

Under ambient and refrigerated conditions stored crude stem extracts maintain their pesticidal activity during 3 months.

Genetic resources and breeding

T. crispa and T. glabra are widely distributed and will not easily become endangered, since they prefer sites with secondary vegetation and show strong regeneration capacity. In India, T. cordifolia is a popular medicinal plant and is locally endangered because of extensive collecting from the wild.

Prospects

T. crispa and the non-Malesian T. cordifolia are well investigated medicinal plants with numerous interesting properties which have been confirmed by modern research. It is expected that they will keep their important role in local medicine in South-East Asia, whereas they have good prospects to play a more prominent role in modern medicines in other parts of the world.

The need for safer pesticides lead to a revival of interest in pesticides of plant origin which can be used in integrated pest management. In the Philippines, Tinospora extracts have been evaluated in the field and proved effective.

Literature

Acevedo, R.A., Santos, A.C. & Pabatao, P., 1970. A diterpene from Tinospora rumphii Boerl. Philippine Journal of Science 97(3): 269-275.
Bänziger, H., 1982. Fruit-piercing moths (Lep., Noctuidae) in Thailand: a general survey and some new perspectives. Mitteilungen der Schweizerischen Entomologischen Gesellschaft 55(3-4): 213-240.
Comley, J.C.W., Titanji, V.P.K., Ayafor, J.F. & Singh, V.K., 1990. In vitro antifilarial activity of some medicinal plants. Acta Leidensia 59(1-2): 361-363.
del Fierro, R.S., 1983. Studies on the mutagenicity, clastogenicity and antimutagenicity potential of Tinospora rumphii Boerlage (Family Menispermaceae). Philippine Scientist 20: 1-10.
Forman, L.L., 1986. Menispermaceae. In: van Steenis, C.G.G.J. & de Wilde, W.J.J.O. (General editors): Flora Malesiana. Series 1, Vol. 10. Kluwer Academic Publishers, Dordrecht, the Netherlands. pp. 188-201.
Forman, L.L., 1991. Menispermaceae. In: Smitinand, T. & Larsen, K. (Editors): Flora of Thailand. Vol. 5(3). The Forest Herbarium, Royal Forest Department, Bangkok, Thailand. pp. 359-365.
Noor, H. & Ashcroft, S.J.H., 1989. Antidiabetic effects of Tinospora crispa in rats. Journal of Ethnopharmacology 27(1-2): 149-161.
Noor, H., Hammonds, P., Sutton, R. & Ashcroft, S.J.H., 1989. The hypoglycemic and insulinotropic activity of Tinospora crispa studies with human and rat islets and HIT-T15 B cells. Diabetologia 32(6): 354-359.
Pachaly, P. & Adnan, A.Z., 1989. The structure of tinocrisposide, a bitter furanoditerpene glucoside from Tinospora crispa. Planta Medica 55(7): 632.
Wadood, N., Wadood, A. & Shah, S.A.W., 1992. Effects of Tinospora cordifolia on blood glucose and total lipid levels of normal and alloxan-diabetic rabbits. Planta Medica 58(2): 131-136.

Selection of species

Authors

Umi Kalsom Yusuf, S.F.A.J. Horsten & R.H.M.J. Lemmens