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Phytochemical, pharmacological evaluation and standardization of selected medicinal plants for anti-inflammatory and anti-arthritic activity

Phytochemical, pharmacological evaluation and standardization of selected medicinal plants for anti-inflammatory and anti-arthritic activity
Rakesh Kumar

2015

Chitkara College of Pharmacy, Chitkara University, Rajpura 140 401, INDIA.

ABSTRACT

The present thesis entitled “Phytochemical, Pharmacological Evaluation and Standardization of Selected Medicinal Plants for Anti-inflammatory and Antiarthritic activity”. Materials and Methods: The Murraya koenigii roots collected from Chitkara University and Ficus lacor aerial roots collected from Panchkula, Haryana plants and identified and authenticated by Dr. H. B. Singh, Head, raw material and herbarium, NISCAIR, Delhi. Results and Discussion: The phytochemical and pharmacognostical evaluation of Murraya koenigii roots were performed. The root is a typical root and in transverse section it shows the features of a dicot root i.e. Epidermis, Cortex, Endodermis, Phloem and Xylem. The microscopy of the powder revealed the presence of xylem vessel, fibre, parenchymatous cell and cork cells. Total ash, acid insoluble ash, water insoluble ash and sulphated ash were 11.25%, 10.57%, 8.75%, 7.025% respectively. The extractive values i.e. petroleum ether, chloroform, ethyl acetate, ethanol and aqueous extract were3.5 %, 5.3 %, 3.2%, 8.5%, 4.5%. The fibre content was 7.53%. The plant can be used as bitter as its bitterness was found to be 2.5 unit/g. The plant possesses haemolytic activity. The plant extracts were good to be free of microbial contamination. The tannin content was 19. The alcoholic and aqueous extracts were screened for presence of amino acid and carbohydrates. The extracts showed the presence of two amino acids viz. Phenylalanine and glycine and two carbohydrates i.e. galactose, ribose and fructose. The preliminary phytochemical screening of Pet. Ether extract, Ethyl acetate extract, Chloroform extract, Ethanol extract, Aqueous extract was performed. The presence of alkaloids, flavonoids, carbohydrates, and sterol in various extracts were observed. This is first ever pharmacognostical study carried out on the aerial roots of Murraya koenigii. Three compounds isolated (i.e. 9-carbethoxy-3-methyl carbazole, 3-methyl carbazole and Koenoline) using different solvent in column chromatography, but these were already reported compounds. MKPE, MKEA and MKCF extracts from Murraya koenigii, both MKPE, MKCF and MKAF-2 were found to be significant in reducing inflammation similar to standard drug in different animal models of inflammation. MKEA was however no significant results showed. MKPE, MKCF and MKAF-2 showed statistically significant inhibition of arthritic lesions (p<0.05) from day 16, (p<0.01) from day 20 and (p<0.001) from day 21 onwards. The extracts administered in higher doses reduced the lesions to a greater extent showing a dose dependent decrease in lesions. The extracts MKPE and MKCF showed significant increase in body weight (p<0.001) as compared to arthritic control group and increase in liver weight (p<0.01), decrease in liver weight (p<0.001) and increase in spleen weight (p<0.001) in arthritis control. MKEA not showed any significant result in body and organ weight estimation. The extracts MKPE and MKCF showed significant decrease in WBC count (p<0.001), increase in hemoglobin contents and RBC counts as compared to control group. The extracts MKPE and MKCF showed significantly (p<0.001) decreased the level of acid phosphatase in blood, whereas they also significantly (p<0.001) decreased the level of acid phosphatase in liver and kidney. MKEA was not able to produce a significant effect. There was significant difference between model group and control group in IL-1 and TNF-α level. After treatment with MKPE, MKEA and MKCF, the level of IL-1 and TNF-α in serum was lower than model group in MKPE and MKCF extracts. In the histological studies, the joints showed the destruction of inflamed joints and continued migration into the synovium and joint fluid of polymorphonuclear leukocytes, lymphocytes and monocytes/macrophages, all of which produce inflammatory cytokines. Thus, pharmacological inhibition of this leukocyte migration and accumulation in arthritis may have beneficial effects for joint preservation. In this study, histological interpretation supported that the administration of MKPE, MKCF (dose100mg/kg body weight) may be protective by decreasing the leukocyte migration. The alcohol extract of roots have shown very significant total phenolic content, total flavonoids content and free radical scavenging activity by DPPH and NO methods.

The pharmacognostical investigations i.e. morphological study of the Ficus lacor aerial root of the plant was performed. The aerial roots are typical roots and in transverse section it shows the features of a dicot root. The microscopy of the powder revealed the presence of annular xylem vessel, lignified fibre, parenchymatous cell and cork cells. Total ash, acid insoluble ash, water insoluble ash and sulphated ash were 14.15%, 8.57%, 10.75%, 6.00% respectively. The extractive values i.e. petroleum ether, chloroform, ethyl acetate, ethanol and aqueous extract were 5.7%, 10%, 5.5%, 4.5%, 10.5%. The fiber content was 9.45%. The plant can be used as bitter as its bitterness was found to be 1.9 unit/g. The foaming index was 124.6. The plant possesses haemolytic activity. The tannin content was 22. The alcoholic and aqueous extracts were screened for presence of amino acid and carbohydrates. The extracts showed the presence of three amino acid viz. alanine, methionine, ornithine and tyrosine and three carbohydrates i.e. galactose, lactose and sucrose. The preliminary phytochemical screening of Pet. Ether extract, Ethyl acetate extract, Chloroform extract, Ethanol extract, Aqueous extract was performed. The presence of flavonoids, carbohydrates, saponins, phenolic compounds, and sterol in various extracts were observed. The three triterpenoidal phytoconstituents isolated (i.e. β- amyrin palmitate, β-amyrin stearate and Myristyl tetradecanoate) using different solvent in column chromatography. These copounds are first time reported in this plant. FLPE, FLEA, FLCF and FLET extracts from Ficus lacor aerial roots, both FLPE, FLET and FLTP-2 were found to be significant in reducing edema as similar to standard drug. Standard drugs produced more significant inhibition of edema. FLPE, FLET and FLTP-2 showed statistically significant inhibition of arthritic lesions (p<0.05) from day 16, (p<0.01) from day 20 and (p<0.001) from day 21 onwards. The extracts administered in higher doses reduced the lesions to a greater extent showing a dose dependent decrease in lesions. The terpenoidal fraction (FLTP-2) showed maximum inhibition as compared with control groups. The extracts FLPE and FLET showed significant increase in body weight (p<0.001) as compared to arthritic control group and increase in liver weight (p<0.01), decrease in liver weight (p<0.001) and increase in spleen weight (p<0.001) in arthritis control. FLEA did not show any significant result in body and organ weight estimation. The extracts FLPE and FLET showed significant decrease in WBC count (p<0.001), increase in hemoglobin contents and RBC counts as compared to control group. The extracts FLPE and FLET showed significantly (p<0.001) decreased the level of acid phosphatase in blood, whereas they also significantly (p<0.001) decreased the level of acid phosphatase in liver and kidney. FLEA and FLCF were not able to produce a significant effect. In the histological studies, interpretation supported that the administration of FLPE and FLET (dose 100mg/kg body weight) may be protective by decreasing the leukocyte migration. The alcohol extract of Ficus lacor aerial roots have shown very significant total phenolic content, total flavonoids content and free radical scavenging activity by DPPH and NO methods.

On the basis of above studies, it may be concluded that both plants are used empirically for the treatment of inflammation and arthritis in folk medicine. There is no data in literature about pharmacognostical, phytochemical and pharmacological potential of plants fractions.