Tylophora Indica Research Articles

Studies on Nootropic Activity of Roots of Tylophora indica in Mice

The present work was designed to assess the potential of aqueous extract of Tylophora indica on cognitive functions in mice using elevated plus maze model and Morris water maze model. Piracetam (200 mg/kg) was used as a standard nootropic agent for comparison. The results of elevated plus maze model showed that, Tylophora indica extract treated mice at 250 mg/kg and 500 mg/kg, significantly (p<0.01) reduced the time required to find the closed arm compared to untreated mice. Further, the extract treatment at 250 mg/kg and 500 mg/kg, showed significant (p<0.01) improvement in scopolamine-impaired performance with respect to acquisition and retention of memory in spatial and working memory tests, in Morris water maze model. In conclusion, Tylophora indica appears to be a promising candidate for improving memory and it would be worthwhile to explore the potential of this plant in management of dementia and Alzheimer's disease.

Effect of Methanolic Extracts of &lt;I&gt;Tylophora indica L.&lt;/I&gt; Leaves on Lipid Profile in Rats

Tylophora indica L is important medicinal plant ofAyurvedic system of medicine and used as home remedy as cardioprotective agent without any scientific background. In this investigation an attempt was made to study the effect of methanolic extract of Tylophora indica L (HETI) on serum lipid profile. Albino rats were administered HETI (100 mg/kg&500 mg/kg) once daily orally for 30 days. Accordingly, serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were among the parameters investigated. We also evaluated the activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels as a measure of hepatic cell damages. It was found in our study that the administration of both doses of HETI to animals fed with high fat diet (HFD) lowered their serum TC, TG and LDL-C significantly, while raised the serum HDL-C. Treatment with HETI reduced serum ALT and AST levels compared to HFD groups. Further, it was also observed that the high dose of HETI significantly decreases serum TC, TG and LDL-C and increases HDL-C in animals fed with normal fat diet (NFD). Based on these data, it is concluded that the chronic administration of HETI in high dose up to 500 mg/kg has definite cardioprotective potential and these results constitute a valid scientific basis for consuming Tylophora indica L . for medicinal application.

Hepatoprotective activity of the methanolic extract ofTylophora indica(Burm. f.) Merill. leaves

The methanolic extract of Tylophora indica leaves was screened for hepatoprotective activity in carbon tetrachloride induced hepatotoxicity in albino rats. The degree of protection was measured by estimating biochemical parameters like Serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase, total protein and level of serum bilirubin (both total and direct). Hepatoprotective activity of methanolic extract at a dose of 200 mg/kg and 300 mg/kg body weight, i.p., was compared with Silymarin (25 mg/kg, i.p.) treated animals. Tylophora indica leaves (200 and 300 mg/kg) exhibited significant reduction in serum hepatic enzymes when compared to rats treated with carbon tetrachloride alone. Furthermore, histopathological studies were also done to support the study. Key words: Carbon tetrachloride, hepatoprotective activity, Silymarin, Tylophora indica

Growth Factor-Induced Vascular Smooth Muscle Cell Proliferation is Inhibited by Tylophorine

Tylophorine, a phenanthroindolizidine alkaloid, is the main active component of Tylophora indica (Asclepiadaceae) which is used in ayurvedic medicine to treat various alleTylophora indica (Asclepiadaceae)rgic and inflammatory disorders, including bronchial asthma, rhinitis, whooping cough and catarrh.

An efficient micropropagation system for Tylophora indica: an endangered, medicinally important plant

An efficient protocol is described for the rapid in vitro multiplication of an endangered medicinal plant, Tylophora indica (Burm. f.) Merrill, via enhanced axillary bud proliferation from nodal explants collected from young shoots of a two-year-old plant. The physiological effects of growth regulators [6-benzyladenine (BA), kinetin (Kin) thidiazuron (TDZ), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or α-naphthalene acetic acid (NAA)], ascorbic acid (AA), different strengths of Murashige and Skoog (MS) medium and various pH levels on in vitro morphogenesis were investigated. The highest number (8.6 ± 0.71) of shoots and the maximum average shoot length (5.2 ± 0.31 cm) were recorded on MS medium supplemented with 2.5 μM BA, 0.5 μM NAA and 100 mg/l AA at pH 5.8. Rooting was best achieved on half-strength MS medium augmented with 0.5 μM IBA. The plantlets regenerated in vitro with well-developed shoot and roots were successfully established in pots containing garden soil and grown in a greenhouse with a 90% survival rate. The regenerated plants did not show any immediate detectable phenotypic variation. The described method can be successfully employed for large-scale multiplication and long-term in vitro conservation of T. indica.

Hepatoprotective activity of alcoholic and aqueous extracts of leaves of Tylophora indica (Linn.) in rats

<b>Objective: </b> To investigate the hepatoprotective activity of alcoholic (ALLT) and aqueous (AQLT) extracts of leaves of <i> Tylophora indica</i> (asclepiadaceae) against ethanol-induced hepatotoxicity. <b> <br>Materials and Methods:</b> Leaf powder of <i> Tylophora indica</i> was successively extracted with alcohol and water. Preliminary phytochemical tests were done and the LD50 values for both extracts determined. The hepatoprotective activity of the ALLT and AQLT were assessed in ethanol-induced hepatotoxic rats. <b> <br>Results:</b> The ALLT showed presence of alkaloids, carbohydrates, steroids, saponins and triterpenes, while alkaloids, carbohydrates and saponins were present with AQLT. The ALLT did not produce any mortality even at 5000 mg/kg while LD50 of AQLT was found to be 3162 mg/kg. Ethanol produced significant changes in physical (increased liver weight and volume), biochemical (increase in serum alanine transaminase, aspartate transaminase, alkaline phosphatase, direct bilirubin, total bilirubin, cholesterol, triglycerides and decrease in total protein and albumin level), histological (damage to hepatocytes) and functional (thiopentone-induced sleeping time) liver parameters. Pretreatment with ALLT or AQLT extract significantly prevented the physical, biochemical, histological and functional changes induced by ethanol in the liver. <b> <br>Conclusion:</b> The present study indicates that ALLT and AQLT extracts possessed hepatoprotective activity. The alcoholic extract was found to exhibit greater hepatoprotective activity than the aqueous extract.

Regeneration of plants from alginate-encapsulated shoots of Tylophora indica (Burm. f.) Merrill, an endangered medicinal plant

SummaryA protocol has been developed for plant regeneration from encapsulated nodal segments of Tylophora indica. Nodal segments collected from a 2-year-old plant were encapsulated in calcium alginate beads. The best gel complex was achieved using 3% (w/v) sodium alginate and 100 mM CaCl2.2H2O. The maximum frequency (91%) of conversion of encapsulated beads into plantlets was achieved on Murashige and Skoog (MS) medium containing 2.5 µM 6-benzyladenine (BA) and 0.5 µM αnaphthalene acetic acid (NAA) after 6 weeks of culture. Encapsulated nodal segments stored at 4°C for 1 – 8 weeks also showed successful conversion, followed by development into complete plantlets when returned to regeneration medium. Conversion of encapsulated nodal segment into plantlets also occurred when the calcium alginate beads were sown directly into SoilriteTM moistened with ½2MS salts. Plants regenerated from encapsulated nodal segments could be hardened, acclimatised and established in soil, with a success rate of 90%. Encapsulation of vegetative propagules in calcium alginate beads could be useful for the exchange, storage and micropropagation of germplasm of this endangered medicinal plant.

Anti-stress, anxiolytic and nootropic activity of roots of Tylophora indica

Anti-stress, anxiolytic and nootropic activity of roots of Tylophora indica

Screening the antiviral activity of Indian medicinal plants against white spot syndrome virus in shrimp

Twenty species of Indian traditional medicinal plants were selected for this study. They were Aegle marmelos, Allium sativum, Aristolochia indica, Azadirachta indica, Cassia fistula, Catharanthus roseus, Curcuma longa, Cynodon dactylon, Lantana camara, Melia azedarach, Mimosa pudica, Momordica charantia, Morus alba, Ocimum americanum, Phyllanthus amarus, Phyllanthus emblica, Psidium guajava, Solanum nigrum, Tridax procumban and Tylophora indica. These plants were used to extract antiviral substances with petroleum ether, benzene, diethyl ether, chloroform, ethyl acetate, methanol, ethanol and distilled water separately using a Soxhlet apparatus. Each extract was tested for its activity against WSSV in marine shrimp and fresh water crabs. Only five plants among these twenty ( A. marmelos, C. dactylon, L. camara, M. charantia and P. amarus) showed antiviral activity against WSSV. The aqueous extract of C. dactylon showed strong antiviral activity at the concentration of 100 mg/kg of animal body. The methanol extract of M. charantia showed significant antiviral activity at the concentration of 150 mg/kg of animal body weight. The aqueous extract of L. camara and P. amarus and methanol extract of A. marmelos showed partial antiviral activity at the concentration of 150 mg/kg of animal body weight.

Effect of Sugars, Gibberellic Acid and Abscisic Acid on Somatic Embryogenesis in Tylophora indica (Burm. f.) Merrill

The effect of sugars, gibberellic acid (GA 3) and abscisic acid (ABA) on somatic embryogenesis from inter-nodal explant-derived callus of Tylophora indica(Burm. f.) Merrill has been investigated. Embryogenic calli were produced from inter-nodal explants and the best result was achieved using Murashige and Skoog (MS) medium supplemented with 4 μmol/L 2, 4-Dichlorophenoxyacetic acid (2, 4-D). Up to 69% of such embryogenic calli differentiated into somatic embryos with an average of 25 embryos per explant (per gram of the calli) on MS medium containing 6 μmol/L kinetin (Kn). The individual effect of sucrose and glucose together with 6 μmol/L Kn was evaluated. There was a significant difference among the concentrations of sugar and among the kinds of sugar tested in somatic embryogenesis. Sucrose at 200 mmol/L with 6 μmol/L Kn gave rise to a maximum embryogenesis (71%) with an average of 49 embryos per explant. However, glucose together with 6 μmol/L Kn or a combination of glucose, sucrose, and 6 μmol/L Kn reduced the percentage of embryogenesis culture and the number of embryos per explant. The presence of GA 3 and ABA at particular concentrations promoted somatic embryogenesis in T. indica. The addition of 10 μmol/L GA 3 into the 200 mmol/L sucrose-containing medium gave a 98% embryogenesis response with an average of 51 embryos per explant. Somatic embryogenesis was significantly enhanced by the addition of 2 μmol/L ABA to 200 mmol/L sucrose-containing medium. On this medium, 95% embryogenesis with an average of 44 embryos per explant was observed. The study reported here indicates that 200 mmol/L sucrose with 6 μmol/L Kn, 200 mmol/L sucrose with 10 μmol/L GA 3, and 200 mmol/L sucrose with 2 μmol/L ABA significantly improved somatic embryogenesis in T. indica, whereas glucose alone or in combination with sucrose had an inhibitory role. The embryos obtained developed normally and transformed into plants easily.

Spontaneous plant regeneration in transformed roots and calli from Tylophora indica: changes in morphological phenotype and tylophorine accumulation associated with transformation by Agrobacterium rhizogenes

We examined the effects of genetic transformation by Agrobacterium rhizogenes on the production of tylophorine, a phenanthroindolizidine alkaloid, in the Indian medicinal plant, Tylophora indica. Transformed roots induced by the bacterium grew in axenic culture and produced shoots or embryogenic calli in the absence of hormone treatments. However, hormonal treatment was required to regenerate shoots in root explants of wild type control plants. Transformed plants showed morphological features typically seen in transgenic plants produced by A. rhizogenes, which include, short internodes, small and wrinkled leaves, more branches and numerous plagiotropic roots. Plants regenerated from transformed roots showed increased biomass accumulation (350-510% in the roots and 200-320% in the whole plants) and augmented tylophorine content (20-60%) in the shoots, resulting in a 160-280% increase in tylophorine production in different clones grown in vitro.

Analysis of the structure‐activity relationships between phenanthroindolizidines and phenanthroquinolizidines

Phenanthroindolizidines and phenanthroquinolizidines are structure-related alkaloids that are well known for their profound cytotoxicity toward a variety of cancer cells and thus have been explored to develop into anti-cancer agents. Tylophorine and cryptoleurine are representative of these two classes of alkaloids respectively. While tylophorine was conceivable to account for the anti-inflammation related effects for Tylophora indica and has been reported to exert significant anti-inflammatory effects in vitro and in vivo, not much known for cryptoleurine and its analogues for anti-inflammation related effects. Although over 70 of phenanthroindolizidine, phenanthroquinolizidine alkaloids, and their analogues have been purified or synthesized, systemic analyses for the structure-activity relationships are needed for more clear elucidation between or among the phenanthroindolizidines and phenanthroquinolizidines. Thus, We synthesized 16 compounds of phenanthroindolizidines (8 compounds, 2 sets) and their counterparts of phenanthroquinolizidines(8 compounds, 2 sets). We analyzed the relationships between the structures and activities of anti-cancer cell growth and anti-inflammation in in vitro systems and also probed the differential activity between and among the phenanthroindolizidines and phenanthroquinolizidines.

Feeding inhibition of Helicoverpa armigera (Hubner) by Eucalyptus camaldulensis and Tylophora indica extracts

Abstract Crude leaf extracts of Eucalyptus camaldulensis and Tylophora indica were evaluated for their antifeedancy against Helicoverpa armigera larvae in no-choice bioassays. Cabbage leaf-disc dipped in different concentrations of leaf extracts was used for feeding fifth instars of H. armigera for 24 hours. Ethanol extracts from both the plant species exhibited significant feeding inhibition activity. The effective concentrations of ethanol extracts that caused 50% reduction in larval feeding (EI50) were 6.9% for E. camaldulensis and 2.8% for T. indica, while these values of hexane extracts were 9.3% and 5.2%, respectively. Compared to the crude leaf extracts, crude alkaloids from T. indica and crude tannins from E. camaldulensis were more potent in reducing larval feedings.

In vitro regeneration and plant establishment of Tylophora indica (Burm. F.) Merrill: Petiole callus culture

Abstract A protocol has been developed for high-frequency shoot regeneration and plant establishment of Tylophora indica from petiole-derived callus. Optimal callus was developed from petiole explants on Murashige and Skoog basal medium supplemented with 10 μM 2,4-dichlorophenoxyacetic acid +2.5 μM thidiazuron (TDZ). Adventitious shoot induction was achieved from the surface of the callus after transferring onto shoot induction medium. The highest rate (90%) of shoot multiplication was achieved on MS medium containing 2.5 μM TDZ. Individual elongated shoots were rooted best on half-strength MS medium containing 0.5 μM indole-3-butyric acid (IBA). When the basal cut ends of the in vitro-regenerated shoots were dipped in 150 μM IBA for 30 min followed by transplantation in plastic pots containing sterile vermiculite, a mean of 4.1 roots per shoot developed. The in vitro-raised plantlets with well-developed shoot and roots were successfully established in earthen pots containing garden soil and grown in a gr...

An efficient in vitro method for mass propagation of Tylophora indica

A protocol of high frequency shoot organogenesis and plant establishment from stem derived callus has been developed for Tylophora indica (Burm. f.) Merrill. - an endangered medicinal plant. Callus was developed on Murashige and Skoog (MS) medium supplemented with 10 μM 2,4,5-trichlorophenoxy acetic acid (2,4,5-T). Multiple shoot induction was achieved from the surface of the callus after transferring onto shoot induction medium. The highest rate (80 %) of shoot multiplication was achieved on MS medium containing 5.0 μM kinetin. The developed shoots rooted best on half-strength MS medium supplemented with 0.5 μM indole-3-butyric acid (IBA). The in vitro raised plantlets with well developed shoot and roots were acclimatized successfully and grown in greenhouse.

Thidiazuron-induced high-frequency shoot organogenesis from leaf-derived callus of ia medicinal climber, Tylophora Indica (Burm. F.) merrill

Tylophora indica (Burm. f.) Merrill is a threatened medicinal climber distributed in the forests of northern and peninsular India. An efficient and reproducible protocol for high-frequency callus regeneration from immature leaf explants of T. indica was developed. Organogenic callus formation from immature leaf pieces was obtained by using Murashige and Skoog (MS) medium supplemented with 7 μM 2,4-dichlorophenoxyacetic acid and 1.5 μM 6-benzyladenine. On this medium 92% explants produced callus. The optimal hormone combination for plantlet regeneration was 8 μM thidiazuron, at which shoot regeneration was obtained from 100% of the cultures, with an average of 66.7 shoots per culture. Histological studies of the regenerative callus revealed that shoot buds were originated from the outermost regions. For root formation, half-strength MS medium supplemented with 3 μM indole-3-butyric acid was used. Plants were transferred to soil, where 92% survived after 3 mo. of acclimatization.

Genetic transformation of Tylophora indica with Agrobacterium rhizogenes�A4: growth and tylophorine productivity in different transformed root clones

We have developed an efficient transformation system for Tylophora indica, an important medicinal plant in India, using Agrobacterium rhizogenes strains LBA9402 and A4 to infect excised leaf and stem explants and intact shoots at different sites. The induction of callus and transformed roots was dependent on the bacterial strain, explant type and inoculation site used. Transformed roots were induced only in explants infected with A. rhizogenes strain A4, while an optimal transformation frequency of up to 60% was obtained with intact shoots inoculated at the nodes. The presence of the left-hand transferred DNA (T(L)-DNA) in the genome of T. indica roots induced by A. rhizogenes was confirmed by PCR amplification of the rooting locus genes of A. rhizogenes. Root growth and the production of tylophorine, the major alkaloid of the plant, varied substantially among the nine root clones studied. Both parameters increased over time in liquid cultures, with maximum biomass and tylophorine accumulation occurring within 4-6 weeks of growth in fresh medium. Interestingly, in liquid culture, the culture medium also accumulated tylophorine up to concentrations of 9.78+/-0.21 mg l(-1).

The root: a potential new source of competent cells for high-frequency regeneration in Tylophora indica.

The purpose of this study was to develop a new micropropagation system for Tylophora indica, an important medicinal plant in India, using root explants as starting material. Root explants cultured on MS medium supplemented with 6-benzyladenine (BA) or 2-isopentyladenine (2iP) produced organogenic nodular meristemoids (NMs) within 4 weeks. NMs induced from the cut ends of root segments showed two types of organogenic response--direct shoot bud formation and somatic embryogenesis--when maintained on induction medium. In 42% of the explants, NMs developed shoot buds directly in the presence of 10.72-26.80 microM BA. On average, 18.5 +/- 0.7 shoots per gram of NM tissue were obtained after each 4-week subculture. Elongation of microshoots and root initiation were correlated with the auxin used, with the optimal response occurring in the presence of 28.54 microM indole-3-acetic acid. In 39% of the explants, NMs dedifferentiated into friable embryogenic callus (FEC) in the presence of BA or 2iP after 12 weeks of culture. Of the different treatments, MS medium supplemented with 10.72 microM BA was the most effective in inducing FEC and somatic embryogenesis: at this concentration 64% of the cultured NMs developed FEC and, on the same medium, 89% of the FEC produced globular somatic embryos (SEs). FEC biomass increased nearly five-fold with every 4-week subculture, and about 30 SEs were recovered per gram of FEC during this period. The best conversion of mature SEs to complete plantlets was obtained on basal MS medium-42%. Plantlets derived via somatic embryogenesis and shoot organogenesis were successfully hardened (88-96%) and transferred to the field.

Rapid mass propagation of Tylophora indica Merrill via leaf callus culture

An efficient protocol has been developed for rapid mass propagation of Tylophora indica from leaf derived callus. Optimal callus was developed from leaf explants on Murashige and Skoog (MS) basal medium supplemented with 10 μ 2,4,5-T. Adventitious shoots were regenerated (85%) from the surface of the callus on MS medium supplemented with 5 μM Kinetin. Individual elongated shoots were rooted on half-strength MS medium containing 0.5 μM IBA. Regenerated plantlets with well developed shoots and roots were successfully transferred to soil. The study demonstrated a dedifferentiated callogenic propagation route via adventitious shoot development in T. indica, which could be useful for large scale multiplication of this endangered medicinal plant.

Plant regeneration through somatic embryogenesis and rapd analysis of regenerated plants in Tylophora indica (Burm. F. Merrill.)

A procedure for the regeneration of complete plantlets of Tylophora indica from cultured leaf callus via somatic embryogenesis is described. Callus induction from leaf explants was on Murashige and Skoog (MS) medium with different concentrations of 2,4-dichlorophenoxyacetic acid (2.4-D; 0.03–3 mg l−1; 0.0–13.56 μM) and kinetin (Kn; 0.01 mg l−1; 0.05 μM). The best response for callus induction was obtained on MS medium containing 2 mg l−1 (9.04 μM) 2.4-D and 0.01 mg l−1 (0.05 μM) Kn. After two subeultures on the same medium the embryogenic callus was transferred to MS medium with different concentrations of the cytokinin, 6-benzyladenine (0.5–3 mg l−1; 2.22–13.32 μM) and 2-isopentenyladenine (2ip; 0.53 mg l−1; 2.46–14.76 μM) along with 0.01 mg l−1 (0.05 μM) indole-3-butyric acid (IBA) for somatic embryo development and maturation. MS medium with 2 mg l−1 (9.84 μM) 2ip produced the maximum number of mature somatic embryos. The mature embryos were bipolar and on transfer to MS basal medium produced complete plantlets. After hardening the regenerants were planted in the Gudalur forests of Western Ghats. Total DNA was extracted from 14 regenerants and the mother plant. Random amplified polymorphic, DNA (RAPD) analysis was carried out using 20 arbitrary oligonucleotides. The amplification products were monomorphic among all the plants revealing the genetic homogeneity and true-to-type nature of the regenerants.