Rajeshkumar et al. World Journal of Pharmacy and Pharmaceutical Sciences. ANTICANCER ACTIVITY OF ANDROGRAPHIS PANICULATA. showing anticancer activity on HEp2 – Human larynx carcinoma cell lines in-vitro KEYWORDS: Andrographis paniculata,Herbal medicine. A refined extract of A. paniculata, also significantly reduces activities of lipid .. The anticancer activity was observed due to the presence of andrographolide.

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This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Salinity causes the adverse effects in all physiological processes of plants. The present study aimed to investigate the potential of salt stress to enhance the accumulation of the anticancer phytochemicals in Andrographis paniculata accessions.

For this purpose, day-old plants were grown in different salinity levels 0. After inducing a period of day salinity stress and before flowering, all plants were harvested and the data on morphological traits, proline content and the three anticancer phytochemicals, including andrographolide AGneoandrographolide NAGand deoxy,didehydroandrographolide DDAGwere measured.

The results indicated that salinity had a significant effect on the aforementioned three anticancer phytochemicals.

In other words, under salinity stress, the tolerant accessions were capable of accumulating the higher amounts of proline, AG, and NAG than the sensitive accessions. Salinity is a major abiotic stress that causes important alterations in the plant growth and development. It may lead to the accumulation or reduction of certain metabolites [ 1 — 3 ]. As an aerial organ, andrographjs are directly exposed to various biotic and abiotic stresses, which demand metabolic adaptations for survival.

Many of those metabolic changes such as sucrose, oligosaccharides, polyhydric alcohols, proline, and polyamines are observable in the leaves of plants exposed to abiotic stresses [ 4 ]. For example, wnticancer level of metabolites was increased in the pabiculata of salt-treated plants such as Arabidopsis thaliana [ 4 ], Oryza sativa [ 5 ], Hordeum vulgareDatura inoxiaGlycine maxTriticum aestivum [ 5 ], and Catharanthus roseus [ 6 ].

Activith in certain amino acids including prolinesugars including sucrose, fructose, and glucoseand polyols including inositols have been reported in some plant species like grapevine [ 7 ], Limonium latifolium [ 8 ], and Lotus japonicas [ 9 ]. Since metabolic adaptations towards environmental stress including salinity affect global metabolic fluxes, some beneficial secondary metabolites of therapeutic importance or so-called phytochemicals would also be affected [ 11 ].

Plant species react against the environmental changes by producing some secondary metabolites such as soluble flavonoids, which protect the cellular structures from oxidative damage and osmotic stress [ 12 ].

In fact, a positive correlation between metabolite enhancement flavonoid and proline and relative water content RWC was reported in rice [ 13 ], reiterating the notion that metabolic adaptations are part of a complex mechanism to overcome threads from stresses. Therefore, the stress conditions have a strong impact on the responsible metabolic pathways for the accumulation of the related natural products [ 11 ]. However, improving the content of some active compounds such as flavonoids could be achieved under abiotic stress condition as an agronomical approach [ 1214 ] and biotechnological approaches genetic transformation [ 15 ] on a large scale in plants.

The plant extract contains three major groups of phytochemical compounds, namely, diterpenes, flavonoids and stigmasterols [ 16 ]. Diterpene lactones are phytochemical structures, which could be found in different parts of this medicinal herb. Among these phytochemicals, three diterpene lactones aandrographis of andrographolide AGneoandrographolide NAGand deoxy,didehydroandrographolide DDAG possess the highest bioactivity in treating the hardly curable diseases [ 1718 ].

The herb exhibited a wide scope of pharmaceutical properties such as anti-HIV [ 19 ], anti-H1N1 [ 20 ], anticancer [ 21 ], antihepatitis [ 22 ], anti-inflammatory [ 23 ], blood purifier, and antidiarrhea [ 24 ]. These compounds are produced mainly from aerial organs of the plant, especially leaves.

However, od production of the three phytochemicals in A. It is assumed that the accumulation of these phytochemicals in A. Therefore, the aim of this study was to determine the changes in the contents of the three main anticancer phytochemicals AG, NAG, and DDAG and their physiological implications in salt tolerance of A. In the current study, phytochemical analysis was conducted on the four different accessions of A.


Here, we report how salt stress enhances the production of the key phytochemicals in the plant. Variations due to salinity levels SL and accessions AC were highly significant.

The reduction in sensitive accessions in terms of all studied morphological traits was higher than in tolerant accessions Figure 1. The decreasing trend of all studied morphological traits was linear Table 2. The dry weight was used to monitor the physiological response of each accession to salinity. After four weeks, a significant decrease in growth driven by salinity was apparent in all accessions. An obvious reduction in TDW was seen in accession and this was taken as a symptom of salt sensitivity of this accession Table 3.

Accordingly, the growth of this accession was stopped after three weeks. By contrast, among the accessions, the highest TDW The analysis of variance showed that different levels of salinity affected the proline and the three main phytochemicals of A. The results showed that proline, AG, and NAG contents were positively correlated to the substrate concentration of NaClwhile the total crude extract amount was negatively correlated to salinity levels Figure 2.

Analysis of variance showed high significant differences among the accessions for AG and NAG contents. The concentration of the AG, the most important cytotoxic principle in A. The broad-sense heritability was calculated for all studied characteristics.

The heritability of proline was the highest 0. This can be utilized as a potential in the next breeding programs to develop the salt-tolerant varieties in A. There were strong relationships between most studied traits. Interestingly, the correlation between proline as well as NAG with the other measured morphological characteristics was significant and positive in the salt-stressed plants.

On the other hand, there was no significant correlation between the three anticancer phytochemicals and the other studied morphological traits, while correlations between AG, NAG, and DDAG were highly significant and positive Table 5. The correlative aspects of the three anticancer phytochemicals anticancerr as well as the accumulation of proline in the tolerant and sensitive A.

Accession showed a growth reduction of dry weight up to By this time, the tolerant accession also had significantly higher proline and phytochemical contents and less ot necrosis than sensitive accession Previous studies have demonstrated that the ability of A. The plants grown under salinity conditions performed different responses after four weeks of salt treatment. Under anyicancer low level of salinity, the salt ions inhibited the biosynthesis of phytochemicals as indicated by the decrease in their amounts in both tolerant and sensitive accessions.

There is a possibility that at the low levels of anticanceg, the plants might be in the adaptive process against salinity stress due to the osmotic reduction of water surrounding the root system. We noticed that the major difference among the tolerant and sensitive accessions was related to their ability in tolerating high salt stress.

At high salinity level, the tolerant plants were able to tolerate the salinity stress paniculaga to the accumulation of osmoregulation and compatible solutes such as proline and soluble flavonoids, while the sensitive accessions were unable to withstand the salinity stress due to the accumulation of sodium ion.

Generally, plants produce secondary metabolites in nature as defense mechanisms under different environmental stresses [ 27 ].

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In this regard, our results indicated that proline as well as the AG and NAG contents is positively correlated with the substrate concentration of salt, in which the tolerant accessions were higher in proline, AG, and NAG contents compared with the sensitive ones. However, we are totally aware that not any correlation does imply causation, yet we believe that in this particular case and according to the previous experiences, our observations are in agreement with the concept of a close relationship between the availability of the secondary metabolites and self-defense systems of the plant species against abiotic stresses.

Likewise, many researchers suggested that proline and soluble flavonoids are involved in osmotic adjustment, protecting the cellular structures from oxidative damage and osmotic stress by accumulation in the vacuole, and so play an important role in increasing the oxidative sctivity tolerance through protecting the chloroplast and photosynthetic systems versus solar radiation by absorbing UV [ 2829 ].

Consequently, the results suggested that increasing the biosynthesis of proline in the plant could protect the cellular structures andrograaphis oxidative damage and osmotic stress. This event complies with the hypothesis that the A. Reportedly, improving the content of some phytochemicals such as flavonoids as one of the most important pharmaceutical contents could be enhanced through salinity and drought stresses [ 121415 ]. As a matter of fact, a part of this enhancement anticancr be due to an obvious reduction panicupata the plant biomass.

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Enhancement of phenolic and flavonoid compounds in onion plant under salinity stress has been reported to improve the deleterious effect of salinity stress [ 34 ].

Accumulation of paniculataa solutes is nontoxic anticaner does not disturb cellular functions even when they are present in high concentrations.

These neutral organic compounds can also improve the inhibitory effects of high ion concentrations on enzymatic activity without interfering with protein structure and function [ 35 ].

Our results indicated a positive correlation between the three anticancer phytochemicals and proline accumulation, which might be closely related to tolerance abilities indicated by physiological performances. This finding matched up with the Chutipaijit et al.

To the best of our reference, the only conducted research related to salt stress in A. In spite of the mentioned trends, the differences between those two accessions were not statistically significant [ 36 ].

To be clearer, it seems that the accessions with high contents of the anticancer phytochemicals under normal condition would deal more successfully with salinity, as well. However, the use of salinity to enhance the biosynthesis of the phytochemicals must be prudentially regarded, and it should not be forgotten that the increased concentration of bioactive compounds such as AG, NAG, and DDAG by salt stress in general is associated with a reduction of biomass production.

Analysis of the Anticancer Phytochemicals in Andrographis paniculata Nees. under Salinity Stress

In other words, the increase of these phytochemicals under salinity condition would be partially compensated by a decline in total biomass. However, this should not be interpreted in the way that saline water and soil are suggested as an alternative solution to increase the production of A.

Fundamentally, it can be concluded that if panculata production of anticancer phytochemicals of A. Solvents AR and chromatography grade for isolation and purification of the compounds were used as supplied by Fisher Scientific Leicestershire, UK. According to Andrgraphis et al. The seeds were germinated as described by Talei et al. The germinated seeds at two initial leaf stages were transferred into the Jiffy media.

The day seedlings were transferred from jiffy media into the pot with sand medium. After 30 days of culturing almost in 70 days oldthe plants were placed in different salinity levels. The experiment was carried out with a split plot based on a randomized complete block design RCBD with two factors and three replicates. These salinities were applied using Since the highest amount of the active components is found just before the plant blooms [ 39 ], the day-old plants were subjected to different salinity levels.

Each plant was irrigated once a day with five levels of saline water. After every three salinity applications, plants were again irrigated with normal Hoagland nutrient solution. The dried materials were chopped into small pieces and ground into fine powder form and were then extracted with a mixture of dichloromethane and methanol DCM: ME at the ratio of 1: The materials were soaked for three days at room temperature.

The process was repeated several times with the same solvent system until the solvent portion becomes colorless. Whatman filter paper no. The solvent extract actiity concentrated under reduced pressure using a rotary evaporator. The concentrated extract was transferred into conical flasks and the remaining solvent was removed.

A final drying was done by placing the concentrated extract in an electric oven at room temperature. One mL of painculata sample was finally filtered into HPLC vials using disposable polypropylene syringe filters with 0. The stock solution was diluted with the same solvent to produce different concentration of working standards 0. One mL of each standard sample was finally filtered into HPLC vials using the same syringe filters in five replicates [ 4041 ].

Calibration curve was generated by linear regression based on peak areas [ 42 ].