ABSTRACT

The observation, identification, description, experimental investigation of the

ingredients and the therapeutic effects of indigenous drugs are all interdisciplinary
field of research. Hence, present research work was aimed at an interdisciplinary
study starting with Pharmacognosy, Phytochemistry and Pharmacological studies.
Based on the extensive medicinal claims of the plants the current research work
was carried out in order to scientifically evaluate the folklore claims and to explore
the unexplored phytochemical constituents since no scientific claim has been made
on anticancer activity in these two plants but evidence of possible use in free
radical mediated diseases has been implicated.
The main objectives of the present study were to collect, identify and confirm the
authenticity of the plant species. To establish pharmacognostical standards and to
perform chemical tests for various phytochemical constituents present in the plant
extracts.To select therapeutically active extract by in-vitro pharmacological studies
and to isolate and identify the structure of the phytoconstituents from the most
active extract.
On the basis of exhaustive literature survey, two plant drugs were selected for the
study: Digera muricata, Amaranthaceae and Cordia dichotoma, Boraginaceae are
ethanobotanically important plant species traditionally used against various
disorders.
The pharmacognostical standards have been established involving both the
botanical parameters as well as physiochemical parameters which include
macroscopy, microscopy, foreign matter, extractive values, ash values, loss on
drying, T.L.C fingerprints of different extractives,Fluorescence analysis,pH value
and Identification of Flavonoids.
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The various parameters of phytochemical analysis for each plant drug

have been done which include preliminary phytochemical screening. The
methanolic extract and fractions obtained were subjected to qualitative chemical
tests for identification of various plant constituents present in the crude drug. The
extract was subjected to preliminary phytochemical investigation for detection of
alkaloids, carbohydrates, glycosides, phenolic compounds, flavonoids, saponins
and sterols. The quantative estimation of phytoconstituents such as Phenolic
content, Flavonoid content, total saponin and total alkaloidal content have been
studied. The phytochemical analysis both the plants resulted in isolation of one
compound each from the methanolic extract. To assess the antioxidant activity of
the methanolic extract and its fractions for the plants under research, an in vitro
assay have been used namely DPPH assay while Cytotoxic potential was
investigated using methanolic extract and fractions of both the plants against HeLa
and A549 cell lines.
Crude methanolic extracts of Digera muricata and Cordia dichotoma were
prepared in methanol by continuous hot soxhlation technique. Crude extract was
fractionated

into

two

organic

and

one

aqueous

fraction.MTT,3-(4,5-

dimethylthiazol-2yl)-2,4 diphenyltetrazolium bromide assay was used to evaluate

the reduction of viability of the cancer cell lines. Morphological analysis was done
by phase contrast microscopy and percentage apoptosis was calculated. Apoptosis
assays using nucleic acid stains namely Propidium iodide (PI) exclusion assay and
Hoestch/PI assay were performed by the help of fluorescence microscopy.
Microscopic fluorescence imaging was used to study the ROS generation in
A549cells after exposure to different concentrations of the methanolic extracts of
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both the plants. The methanolic extract was tested against A549 cells for ROS assay.
The qualitative and quantitative analysis of ROS generation suggests methanol
extract induced reactive oxygen species mediated apoptosis induction in A549 cells.
It is suggested that the further isolation of extract and its fractions may give
a potential compound which may prove to be a world widely acceptable anticancer
remedy. The overall results indicate the promising baseline information for the
potential uses of methanolic extract of both the plants as an anticancer agent.

ACKNOWLEDGEMENTS
On completion of my dissertation work, I would like to express my
wholehearted gratitude to all those who assisted me to reach this
destination.

Firstly,

bow

down

my

head

to

the

Almighty

whose

omnipresence has always guided me and made me energized to carry out

this research work.
Next, I express my deep sense of gratitude and reverence to my Supervisor,
Dr. Arshad Hussain, Associate Professor, Faculty of Pharmacy, Integral
University, Lucknow for his excellent guidance, constant and untiring
supervision, help and encouragement throughout the investigation and
preparation of this manuscript.
I am extremely grateful and indebted to my Co- Supervisor Dr. A.H.A
Farooqui, Professor, Faculty of biotechnology, Integral University, Lucknow for
inspiring suggestions and valuable advice for this investigation.
I wish to extend my sincere thanks to Committee Chairman Prof. (Dr.) H. H.
Siddiqui, Advisor to VC, Faculty of Pharmacy, Integral University, Lucknow for
his guidance and support on this project.
I would also like to acknowledge Lucknow University and Central Drug
Research Institute (CDRI) for providing their help and support in performing
the lab work and characterization of samples.
Special appreciation is extended to my colleagues Ms. Shabana Khatoon, Dr.
Poonam Kushwaha, Dr. Mehnaz Kamal, Mr. Mohd. Arif , Dr. Anuradha Mishra
for their assistance, advice, and encouragement.

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Besides, I cannot forget the favors of Ms.Barnali, Shiva and Mr. Ahmad for
their moral support, help and encouragement throughout the course of this
work.
Lastly, I acknowledge with highest sense of regards, the support, blessings
and love and affection of my parents, my husband, my brother, sister and
other members of my family without which the present investigation would
not have been successful.

SHAZIA USMANI

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