The present study was undertaken to test the chemopreventive effects of one herbal medicinal plant, Indigofera aspalathoides, on chemically. The treatment by I. aspalathoides on fibrosarcoma bearing rats has improved the levels Indigofera aspalathoides (I. aspalathoides) treatment for rat arthritis is. Abstract: Civarvembu/ Sivanimba is botanically equated to Indigofera aspalathoides Vahl ex. DC. (Leguminosae). The plant is prescribed for eczema, psoriasis.
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The present study was undertaken to test the chemopreventive effects of one herbal medicinal plant, Indigofera aspalathoideson chemically induced carcinogenesis in rats. A well-known polyaromatic hydrocarbon, namely, methylcholanthrene, which is a known carcinogenic substance, was used to induce fibrosarcoma in Wistar strain of male albino rats. Fibrosarcoma rats were treated with aqueous extracts of Indigofera aspalathoides.
The indigoffra were divided into four groups, each consisting of six animals. Group I served as normal control, Group II served as fibrosarcoma-induced animals, Group III were fibrosarcoma-bearing animals treated with aqueous extracts of Indigofera aspalathoidesand Group IV animals, which were normal healthy animals treated with Indigofera aspalathoides aqueous extract, served as drug control set.
The fibrosarcoma was proved by pathological examinations. The activity levels of nucleic acids such aspapathoides total DNA and RNA and hexose, hexosamine, and sialic acid in liver and kidney of treated rats were used to monitor the chemopreventive role of the plant extract. The observed increase in the levels of DNA, RNA, hexose, hexosamine, and sialic acid in liver and kidney tissues of fibrosarcoma-bearing animals reached near normal state after the treatment with aqueous extracts of Indigofera aspalathoidessuggesting that Indigofera aspalathoides does have a chemotherapeutic role.
The disease cancer will become the leading cause of mortality among human population [ 1 ]. Chemical carcinogenesis due to a number of agents is a proven fact. Consistent evidence shows carcinogenesis to be caused by certain chemicals such as polycyclic aromatic hydrocarbons PAH like methylcholanthrene or 3-methylcholanthrene. They are readily detected in the air we breathe or the tobacco that we smoke [ 3 ].
The specific type of cancer produced varies with the root of administration and include, tumors of the skin, soft aspalathojdes, and breast. PAHs are metabolized by cytochrome P -dependent mixed-function oxidases to give electrophilic epoxides.
The epoxides either get detoxified or bind covalently to the DNA. The resultant abnormal DNA induces abnormal cell division and produce malignancy [ 3 ].
Carcinogenesis is a multistage disease process. The events leading to cancer is sequential, involving both intrinsic and extrinsic factors. At the molecular level, cancer is caused by abnormal gene expression. This occurs through a number of mechanisms including a direct damage to DNA leading to abnormal transcription of genome.
Majority of chemical carcinogens cause mutations presumably in critical genes and alter the cell growth and differentiation through epigenetic factors.
The development of fully malignant tumor is to involve the activation of altered expression of proto-oncogenes to oncogenes and the loss or inactivation of tumor suppression gene, the function of which is aspalathoided control normal cellular activity [ 4 ]. In the present study, MCA was used as a fibrosarcoma inducing agent in rats. Fibrosarcoma is a tumor composed of aspwlathoides fibers forming mesenchymal cells of fibroblasts aspalathoidess they arise from subcutaneous fibrous tissues [ 5 ].
The tumor is a inditofera, firm, and pale mass without any capsule and infiltrates into the surrounding tissues. The tumor occurs near fibrous tissues and mostly in the region of thigh and knee. The peak age for fibrosarcoma incidence is 30 to 55 years.
Most studies have reported a slightly higher incidence of this tumor in men than in women [ 6 ]. Fibrosarcomas are assigned grades, from 1 to 3, the higher grade sarcomas grade 2 or 3 pose a significant threat and present problems in local control [ 7 ]. Histological grading of fibrosarcoma is mainly based on the degree of cellularity, degree of cellular differentiation, number of mitotic figures, the amount of collagen produced by the tumor cells, and the extent of necrosis.
It is a fact that both surgery and aspalahtoides may fail to cure cancer chiefly because the tumors might have already decimated to other parts of the body. For this reason, chemotherapy is considered as one of the best alternative. Chemotherapy, although being a major treatment modality used in the control of advanced stages of malignancies, exhibits severe toxicity on normal tissues [ 8 ].
Plant products which have antioxidant and anticancer aspalathoises could be considered indivofera good chemopreventive agents. Recent researches revolve around the urgency to evolve suitable chemotherapy with less or no toxic side effects. Plants have played a major role in treatment of various diseases of animals and humans since time immemorial. A large number of active principles from traditional medicinal plants have been indlgofera to have chemopreventive properties [ 10 ].
Most of the studies on chemoprevention are based on individual chemicals with well-defined mechanisms. Since carcinogenesis is a infigofera and multistage process, the individual active compounds or active principles may not always be effective to control all the stages and, therefore, it is of interest to investigate the chemopreventive effect of crude extracts of plants which might control indibofera different stages of cancer etiology as compounds like saponins, tannins, steroids, alkaloids present therein might act holistically to effect a proper cure for cancer.
The plant Indigofera aspalathoides is used as idnigofera potent drug for many diseases in the traditional Indian system. This plant belongs to the family Papilionaceae and it is commonly known as Sivanar Vembu or Iraivanar Vembu in Tamil language of India. The leaves, flowers, and tender shoots are said to be cooling and demulcent [ 11 ]. This plant is used for the treatment of leprosy, syphilis, and various skin diseases [ 12 ]. The plant mainly contains saponins, tannins, steroids, alkaloids, flavonoids, and reducing sugars.
The present study was aimed at finding out the curative role of the aqueous extract of Indigofera aspalathoides on MCA-induced fibrosarcoma in rats.
Some reports of its anticancer and antioxidant properties are already been studied by us [ 13 — 16 ]. The present study is to further know this plant’s chemopreventive role on some more parameters like total DNA and RNA content and on parameters like hexose, hexosamine, and sialic acid.
The extract was filtered and concentrated on a water bath. The inorganic material was precipitated and filtered off. The filtrate was again concentrated in a China dish and dried in vacuum.
This was stored in refrigerator for further and future use.
In case of the death, the limit test was terminated and main test was conducted. If the animal survived, four additional animals were dosed sequentially so that five animals could be tested. However, if three animals died, the limit test was terminated and the main test was performed. If an animal died unexpectedly late in the study and there were other survivors, it was appropriate to stop dosing and observing all animals to see if other animals also die during a similar observation period.
AEIA was found to be safe even at a higher concentration and, based on this, the dose for the chemopreventive activity was chosen. The animals were fed with normal pellet diet rat chew and water ad libitum. The animals were sacrificed by cervical decapitation at the end of the experimental period and the liver and kidney were dissected out and known weight of liver and kidney were homogenized in 0.
Animals were starved overnight before sacrifice. Tumors which appeared in about 4 weeks after implantation were highly localized and were maintained by serial transplantation. The tumor was minced and suspended in normal saline. The transplanted tumor became palpable in 4—6 days time. The liver and kidney of control and experimental animals were used for histopathological analysis. From the figures, it is clear that the treatment of this drug has restored the liver and kidney architecture.
The rats were divided into four different groups, each group consisting of six animals. Tumor measurements were made using a vernier calipers, and tumor diameter Td was calculated using the formula stated elsewhere.
The experiments were repeated twice. The Tukey’s multiple comparison method was used to compare the means of different groups and the significance was denoted by P value.
The method of Burton [ 19 ] was followed for DNA estimation. A reagent blank and standards were also carried out. The estimation was carried out by the method of Rawal et al. The tubes were heated in a water bath for 20 minutes.
Reagent blanks and standards were also treated in the same way. The method of Niebes, [ 22 ], was followed for estimating hexose. The level of Hexosamine was assayed by the method of Wagner, [ 23 ].
Standard indigodera was made up to 1 mL.
File:Indigofera – Wikimedia Commons
The method of Warren, [ 24 ], was used to estimate sialic acid. The hydrolysed materials were neutralized and used for estimation of sialic acid.
After the incubation, the reaction was arrested by the addition of 0. The tubes were shaken well. Once settled, the butanol phase was separated. Standard N-acetylneuraminic acid was also treated similarly. The lack of difference in the results in Group IV and Group I normal control animals clearly demonstrates that Indigofera aspalathoides extracts do not have any toxic side effects which justifies the choice of this drug. Nucleic acids play a vital role during neoplastic transformation.
DNA and RNA contents were studied in order to determine the effects of Indigofera aspalathoides on macromolecular synthesis in fibrosarcoma. It has been observed that the tumor growth corresponds with the elevated levels of DNA and RNA synthesis in the liver and kidney.
But after the treatment with Indigofera aspalathoides, there was a marked decrease in the content of these two nucleic acids suggesting that the plant extract might have controlled their biosynthesis indicating the tumoricidal effect during cancer therapy. Table 2 depicts the levels of hexose, hexosamine, and sialic acid in liver and kidney of control and experimental animals.
But in Group III drug-treated animals, these protein levels were reverted back to near normal condition as compared with Group II fibrosarcoma animals. No significant changes of these levels were observed in Group IV drug-alone-treated animals when compared to normal control group I animals. The levels of hexose, hexosamine, and sialic acid in liver and kidney of control and experimental animals. Heidelberger, [ 25 ], has sited the reduced levels of proteins in neoplastic tissues.
The liver is an important site of protein metabolism and it has the highest rate of protein synthesis. Major protein mass of the organism is severely affected in cancer cachexia. Protein waste implies the underlying metabolic imbalance which is being expressed by an elevation in the apparent protein degradation rate with no change in the apparent synthesis rate [ 26 ].
Reduced liver protein in Moris-hepatoma-bearing animals, in Walker carcinoma, and in some other reports suggests increased protein degradation [ 27 ]. There is decrease in the recycling of amino acids in tumor conditions resulting in enhanced efflux of these amino acids from the tissues.
Thus the host response to the tumor load results in tissue protein breakdown. Treatment with Indigofera aspalathoides might have prevented this protein breakdown thus bringing the protein levels to near normal condition.