{"id":55844,"date":"2024-03-20T11:14:13","date_gmt":"2024-03-20T11:14:13","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=55844"},"modified":"2024-04-01T19:23:39","modified_gmt":"2024-04-01T19:23:39","slug":"assessment-of-anticancer-activity-of-crude-ethanolic-extracts-of-moringa-oleifera-pod-and-leaves-on-712-dimethylbenz-anthracene-induced-skin-cancer-in-mice","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol17no1\/assessment-of-anticancer-activity-of-crude-ethanolic-extracts-of-moringa-oleifera-pod-and-leaves-on-712-dimethylbenz-anthracene-induced-skin-cancer-in-mice\/","title":{"rendered":"Assessment of Anticancer Activity of Crude Ethanolic Extracts of Moringa Oleifera Pod and Leaves on 7,12 – Dimethylbenz Anthracene Induced Skin Cancer in Mice."},"content":{"rendered":"\n

Introduction<\/strong><\/p>\n\n\n\n

Cancer is one of the deadliest diseases and its treatment is always challenging.The incidence of Cancer is expected to rise by around 70%, 20 years from now. According to the Cancer facts and figures 2022, it is predicted that 1.9 million patients are to be newly diagnosed and 6,09,360 deaths due to cancer is likely to occur in the United States.1<\/sup><\/p>\n\n\n\n

Carcinoma of the skin is the commonest cancer in the world.2<\/sup> 1-2% of Cancers in India are skin cancers. There are various modalities of treatment available for the treatment of skin cancers such as surgery, chemotherapy, radiotherapy, immunotherapy but their cost and toxicity are the major concerns.<\/p>\n\n\n\n

To overcome these difficulties researchers are constantly studying the anti-cancer effects of various natural plant sources.3<\/sup>Moringa Oleifera<\/em> commonly called the Drumstick is traditionally used in ayurvedic medicine to treat several diseases. Moringa oleifera <\/em>being an Indian medicinal plant easily cultivatable in the southern Asian continentis a great source of nutritional content with high economic importance.<\/p>\n\n\n\n

Moringa oleifera<\/em> has medicinal values in its every part like leaves, pods, flower, bark and root. Leaves serve as medicine for asthma, dyslipidemia and flowerscan be used in treatment of arthritis.4<\/sup><\/p>\n\n\n\n

Moringa oleifera <\/em>has been found to have diverse medicinal activities and is studied in vitro<\/em> and in vivo<\/em> models of diabetes mellitus, hypertension, peptic ulcer, epilepsy, fever, inflammatory disorders, bacterial and fungal infections.5,6<\/sup><\/p>\n\n\n\n

This study is done to investigate the anti-cancer effect of Moringa oleifera<\/em> pod and leaf extracts in 7,12 \u2013diethylbenzanthracene (DMBA) induced skin cancer in mice.<\/p>\n\n\n\n

Materials and Methods<\/strong><\/p>\n\n\n\n

Animals<\/strong><\/p>\n\n\n\n

30 Swiss Albino mice, 4-6 weeks old and weighing approximately 20-35\ngrams were obtained from the animal house of the Institution. The animals were\nquarantined for one week before the study. Animals were housed in clean\npolypropylene cages availing free access to standard pellet diet and water at\nappropriate temperature with natural day-light cycle. The study procedures were\ndone as per the guidelines of CCSEA. <\/p>\n\n\n\n

Chemicals<\/strong><\/p>\n\n\n\n

7,12 – dimethylbenz anthracene (DMBA), was\npurchased from Sigma-Aldrich Ltd., Bangalore. Acetone was used as a solvent for\nDMBA.5-Fluoro uracil (0.5%), was used as the standard anticancer drug. Ethanol\nwas used for pod and leaf extraction. Normal saline was used as a placebo for\nmice in control group.<\/p>\n\n\n\n

Ethics<\/strong><\/p>\n\n\n\n

The study was conducted after being approved by the Institutional animal\nethics committee.(IAEC2\/Proposal:62\/A.Lr:45\/Dt:10.08.2021)<\/p>\n\n\n\n

Extract preparation<\/strong><\/p>\n\n\n\n

Moringa oleifera <\/em>leaves and pods were gathered from nearby locality and washed thoroughly\nand shade dried for a week. Dried specimen was crushed in a mixer grinder and\nextracted with ethanol for 72 hours. Extracts after filtration was evaporated\ncompletely on water bath. After extraction the residue obtained was dissolved\nin 65% ethanol and centrifuged at -4 degrees at 4000rotations per minute for 10\nminutes and supernatant was used.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 1: Preparation of Pod extract<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 2: Preparation of Leaf extract.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Experimental design<\/strong><\/p>\n\n\n\n

The study had 6 groups with 5 animals in each.<\/p>\n\n\n\n

Table 1: Experimental Design<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

Groups<\/strong><\/p>\n<\/td>\n

\n

Induction + Treatment<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

DMBA + Normal saline (10ml\/kg, p.o) for 21 days<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

DMBA + 0.5% 5-Fluoro uracil (topical application) for 21 days(7)<\/sup><\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

DMBA +Moringa Oleifera <\/em>ethanolic pod extract (MOPE 500mg\/kg p.o) for 21 days<\/p>\n<\/td>\n<\/tr>\n

\n

IV<\/strong><\/p>\n<\/td>\n

\n

DMBA +Moringa Oleifera <\/em>ethanolic pod extract (MOPE 1000mg\/kg p.o) for 21 days<\/p>\n<\/td>\n<\/tr>\n

\n

V<\/strong><\/p>\n<\/td>\n

\n

DMBA +Moringa Oleifera <\/em>ethanolic leaf extract (MOLE 500 mg\/kg p.o) for 21 days<\/p>\n<\/td>\n<\/tr>\n

\n

VI<\/strong><\/p>\n<\/td>\n

\n

DMBA +Moringa Oleifera <\/em>ethanolic leaf extract (MOLE 1000mg\/kg p.o) for 21 days<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

DMBA \u2013 7, 12DimethylBenzanthracene.p.o \u2013 per oral. MOPE \u2013 Moringa Oleifera Pod extract.  MOLE \u2013 Moringa Oleifera Leaf extract.<\/p>\n\n\n

Dose of the extracts in groups III to VI were calculated based on previous study.8<\/sup><\/p>\n\n\n\n

Tumour induction<\/strong><\/p>\n\n\n\n

Tumour was induced as per the standard method suggested in previous literature.9 <\/sup>The dorsal aspect of each mouse was shaved for about 1 inch. 7,12 – dimethylbenz anthracene (DMBA)100\u00b5g, dissolved in 100\u00b5L acetone was used to induce tumour. The solution was painted topically over the shaved area twice weekly for 4 weeks to all animals. The appearance of palpable skin tumour was considered as tumour induction. <\/p>\n\n\n\n

Following the 4th week, treatment was started to each group and continued for next 3 weeks. At the end of 7th week, the study was terminated. 0.3ml blood was collected under anaesthesia for estimation of haematological and biochemical parameters and all animals were euthanized and tumour tissue collected for histopathological examination.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 3: Mice with hair removed on dorsum. Mice with tumour.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

General parameters<\/strong><\/p>\n\n\n\n

Behaviour\nof the mice, movements, food and water intake was observed daily and body\nweight measured at weekly intervals. <\/p>\n\n\n\n

Tumour assessment<\/strong><\/p>\n\n\n\n

Measurement of tumour size was done using the digital vernier calliper of 0.01mm precision with the mice held in hand. Tumour area is body surface area occupied by the tumour calculated by the formula \u03c0\/4*length*width. Length of the tumour indicates the longest diameter along the vertical axis and width indicates the diameter along the horizontal axis.10 <\/sup>Measurements were taken at the end of 4th and 7th week of study. <\/p>\n\n\n\n

Haematological and biochemical parameters<\/strong><\/p>\n\n\n\n

 The Haemoglobin, RBC, WBC and platelet count,\nliver enzymes – SGOT, SGPT, blood urea and serum creatinine were evaluated for\nall animals before the start of the study(baseline) and at completion (at the\nend of 7th week).<\/p>\n\n\n\n

Histopathological examination<\/strong><\/p>\n\n\n\n

After\nstudy completion, euthanasia of animals was done and the tissues (skin with\ntumour, liver and kidney) were sent for histopathological examination. The\ntissues were processed and sectioned in microtome and stained with haematoxylin\nand eosin. Examination of the sectioned slides were done under light\nmicroscopy.<\/p>\n\n\n\n

Statistical Analysis<\/strong><\/p>\n\n\n\n

Statistical\nPackage for Social Sciences v-26was used for statistical analysis. Descriptive\nstatistics was done to obtain the basic characteristics of the data. For the\nintragroup comparison, parametric paired t<\/em> test was used to find the p<\/em>\nvalue. For weight, Graphical representation was used to find the difference\namong the groups.<\/p>\n\n\n\n

Results and Discussion:<\/strong><\/p>\n\n\n\n

Tumour size<\/strong><\/p>\n\n\n\n

There was progressive increase in tumour\nsize in the control group. There was a decrease in tumour size in the groups\nreceiving MOLE and MOPE at different concentration (500mg\/kg and 1000mg\/kg)\nwhich was comparable with the standard treated group. Table 2.<\/p>\n\n\n\n

Table 2: Tumour Size in mm<\/strong><\/p>\n\n\n\n\n\n\n\n
\n

 <\/strong><\/p>\n<\/td>\n

\n

DMBA<\/strong><\/p>\n<\/td>\n

\n

DMBA<\/strong><\/p>\n

+5-FU<\/strong><\/p>\n<\/td>\n

\n

DMBA+ MOPE<\/strong><\/p>\n

(500 mg\/kg)<\/strong><\/p>\n<\/td>\n

\n

DMBA+ MOPE<\/strong><\/p>\n

(1000 mg\/kg)<\/strong><\/p>\n<\/td>\n

\n

DMBA+ MOLE<\/strong><\/p>\n

(500 mg\/kg)<\/strong><\/p>\n<\/td>\n

\n

DMBA+ MOLE<\/strong><\/p>\n

(1000 mg\/kg)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

4th<\/sup> week<\/strong><\/p>\n<\/td>\n

\n

9<\/p>\n<\/td>\n

\n

8.5<\/p>\n<\/td>\n

\n

7.8<\/p>\n<\/td>\n

\n

8.6<\/p>\n<\/td>\n

\n

8.4<\/p>\n<\/td>\n

\n

9<\/p>\n<\/td>\n<\/tr>\n

\n

7th<\/sup> week<\/strong><\/p>\n<\/td>\n

\n

13<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

2<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

1.5<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Body Weight<\/strong><\/p>\n\n\n\n

The weight of mice\nmeasured over regular intervals during the study showed significant changes.\nThe control group showed marked decline in their weight suggesting the effect\nof carcinogenesis. The standard treated group gained weight during the\ntreatment after initial fall. The animals treated with ethanolic extracts of\nMoringa showed similar weight gain after treatment, preceded by loss after the\ninduction.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 4: Body Weight in Grams.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Haematological and Biochemical parameters<\/strong><\/p>\n\n\n\n

Haemoglobin (Hb), total erythrocyte\ncount (RBC), total leukocyte (WBC) count, platelet count, liver enzymes – SGOT,\nSGPT, blood urea and serum creatinine were assessed before tumour induction and\nafter the termination of the study. It was observed that there was a reduction\nin the Hb, RBC, WBC and platelet count and a rise in the liver enzymes and\nrenal parameters in the control group when the baseline (before tumour\ninduction) readings were compared with the week 7 values. The standard\nanti-cancer drug 5-FU and the MOPE and MOLE treated groups showed a rise in the\nhaematological parameters and near normal biochemical parameters at the end of\nthe study which were comparable with the baseline values.<\/p>\n\n\n\n

Table 3: Hb and Platelet Count<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

 <\/strong><\/p>\n<\/td>\n

\n

Hb gm\/dL<\/strong><\/p>\n<\/td>\n

\n

Platelet Count 103<\/sup>\/Cu.mm<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

 <\/strong><\/p>\n<\/td>\n

\n

Baseline<\/strong><\/p>\n<\/td>\n

\n

Week 7<\/strong><\/p>\n<\/td>\n

\n

P value<\/strong><\/p>\n<\/td>\n

\n

Baseline<\/strong><\/p>\n<\/td>\n

\n

Week 7<\/strong><\/p>\n<\/td>\n

\n

P Value<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Control <\/strong><\/p>\n<\/td>\n

\n

13.80\u00b1.556<\/p>\n<\/td>\n

\n

11.14\u00b1.702<\/p>\n<\/td>\n

\n

.004*<\/p>\n<\/td>\n

\n

274.88\u00b135.60<\/p>\n<\/td>\n

\n

138.80\u00b128.83<\/p>\n<\/td>\n

\n

.000*<\/p>\n<\/td>\n<\/tr>\n

\n

Standard(5-FU)<\/strong><\/p>\n<\/td>\n

\n

11.00\u00b1.341<\/p>\n<\/td>\n

\n

14.00\u00b1.353<\/p>\n<\/td>\n

\n

.003*<\/p>\n<\/td>\n

\n

278.80\u00b137.70<\/p>\n<\/td>\n

\n

299.60\u00b122.60<\/p>\n<\/td>\n

\n

.010*<\/p>\n<\/td>\n<\/tr>\n

\n

MOPE-500<\/strong><\/p>\n<\/td>\n

\n

14.48\u00b1.334<\/p>\n<\/td>\n

\n

13.40\u00b1.367<\/p>\n<\/td>\n

\n

.000*<\/p>\n<\/td>\n

\n

264.20\u00b175.85<\/p>\n<\/td>\n

\n

272.40\u00b155.50<\/p>\n<\/td>\n

\n

.039*<\/p>\n<\/td>\n<\/tr>\n

\n

MOPE-1000<\/strong><\/p>\n<\/td>\n

\n

11.10\u00b1.514<\/p>\n<\/td>\n

\n

13.66\u00b1.511<\/p>\n<\/td>\n

\n

.018*<\/p>\n<\/td>\n

\n

272.00\u00b179.90<\/p>\n<\/td>\n

\n

296.60\u00b166.22<\/p>\n<\/td>\n

\n

.010*<\/p>\n<\/td>\n<\/tr>\n

\n

MOLE-500<\/strong><\/p>\n<\/td>\n

\n

14.58\u00b1.221<\/p>\n<\/td>\n

\n

12.22\u00b1.485<\/p>\n<\/td>\n

\n

.009*<\/p>\n<\/td>\n

\n

262.00\u00b112.28<\/p>\n<\/td>\n

\n

278.00\u00b151.20<\/p>\n<\/td>\n

\n

.015*<\/p>\n<\/td>\n<\/tr>\n

\n

MOLE-1000<\/strong><\/p>\n<\/td>\n

\n

13.50\u00b1.393<\/p>\n<\/td>\n

\n

14.44\u00b1.415<\/p>\n<\/td>\n

\n

.004*<\/p>\n<\/td>\n

\n

271.20\u00b171.92<\/p>\n<\/td>\n

\n

290.10\u00b166.69<\/p>\n<\/td>\n

\n

.013*<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

*<\/strong> P value <0.05 indicating Statistical significance<\/p>\n\n\n

 Table 4: RBC Count and WBC Count<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
 <\/td>\n\n

RBC Count million\/Cu.mm<\/strong><\/p>\n<\/td>\n

\n

WBC Count 103<\/sup>\/Cu.mm<\/strong><\/p>\n<\/td>\n<\/tr>\n

 <\/td>\n\n

Baseline<\/strong><\/p>\n<\/td>\n

\n

Week 7<\/strong><\/p>\n<\/td>\n

\n

P Value<\/strong><\/p>\n<\/td>\n

\n

Baseline<\/strong><\/p>\n<\/td>\n

\n

Week 7<\/strong><\/p>\n<\/td>\n

\n

P Value<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Control <\/strong><\/p>\n<\/td>\n

\n

8.28\u00b1.030<\/p>\n<\/td>\n

\n

7.93\u00b1.035<\/p>\n<\/td>\n

\n

.000*<\/p>\n<\/td>\n

\n

5.90\u00b1.578<\/p>\n<\/td>\n

\n

3.66\u00b1.498<\/p>\n<\/td>\n

\n

.006*<\/p>\n<\/td>\n<\/tr>\n

\n

Standard(5-FU)<\/strong><\/p>\n<\/td>\n

\n

8.81\u00b1.087<\/p>\n<\/td>\n

\n

8.94\u00b1.038<\/p>\n<\/td>\n

\n

.039*<\/p>\n<\/td>\n

\n

6.16\u00b1.089<\/p>\n<\/td>\n

\n

6.86\u00b1.378<\/p>\n<\/td>\n

\n

.023*<\/p>\n<\/td>\n<\/tr>\n

\n

MOPE-500<\/strong><\/p>\n<\/td>\n

\n

8.76\u00b1.014<\/p>\n<\/td>\n

\n

8.81\u00b1.032<\/p>\n<\/td>\n

\n

.041*<\/p>\n<\/td>\n

\n

6.44\u00b1.181<\/p>\n<\/td>\n

\n

6.56\u00b1.240<\/p>\n<\/td>\n

\n

.051<\/p>\n<\/td>\n<\/tr>\n

\n

MOPE-1000<\/strong><\/p>\n<\/td>\n

\n

8.82\u00b1.038<\/p>\n<\/td>\n

\n

8.93\u00b1.043<\/p>\n<\/td>\n

\n

.023*<\/p>\n<\/td>\n

\n

6.28\u00b1.286<\/p>\n<\/td>\n

\n

6.64\u00b1.240<\/p>\n<\/td>\n

\n

.036*<\/p>\n<\/td>\n<\/tr>\n

\n

MOLE-500<\/strong><\/p>\n<\/td>\n

\n

8.57\u00b1.043<\/p>\n<\/td>\n

\n

8.64\u00b1.045<\/p>\n<\/td>\n

\n

.051<\/p>\n<\/td>\n

\n

6.65\u00b1.556<\/p>\n<\/td>\n

\n

6.75\u00b1.129<\/p>\n<\/td>\n

\n

.069<\/p>\n<\/td>\n<\/tr>\n

\n

MOLE-1000<\/strong><\/p>\n<\/td>\n

\n

8.85\u00b1.034<\/p>\n<\/td>\n

\n

8.96\u00b1.040<\/p>\n<\/td>\n

\n

.032*<\/p>\n<\/td>\n

\n

6.54\u00b1.867<\/p>\n<\/td>\n

\n

6.88\u00b1.311<\/p>\n<\/td>\n

\n

.047*<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

*<\/strong><\/a> P value <0.05 indicating Statistical significance<\/p>\n\n\n

Table 5: Liver Enzymes: SGOT, SGPT<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
 <\/td>\n\n

SGOT U\/L<\/strong><\/p>\n<\/td>\n

\n

SGPT U\/L<\/strong><\/p>\n<\/td>\n<\/tr>\n

 <\/td>\n\n

Baseline<\/strong><\/p>\n<\/td>\n

\n

Week 7<\/strong><\/p>\n<\/td>\n

\n

P value<\/strong><\/p>\n<\/td>\n

\n

Baseline<\/strong><\/p>\n<\/td>\n

\n

Week 7<\/strong><\/p>\n<\/td>\n

\n

P value<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Control <\/strong><\/p>\n<\/td>\n

\n

58.40\u00b12.31<\/p>\n<\/td>\n

\n

65.40\u00b12.30<\/p>\n<\/td>\n

\n

.000*<\/p>\n<\/td>\n

\n

36.80\u00b14.65<\/p>\n<\/td>\n

\n

40.00\u00b14.74<\/p>\n<\/td>\n

\n

.000*<\/p>\n<\/td>\n<\/tr>\n

\n

Standard<\/strong><\/p>\n<\/td>\n

\n

59.61\u00b12.22<\/p>\n<\/td>\n

\n

60.80\u00b14.32<\/p>\n<\/td>\n

\n

.071<\/p>\n<\/td>\n

\n

35.23\u00b16.49<\/p>\n<\/td>\n

\n

35.21\u00b13.42<\/p>\n<\/td>\n

\n

.063<\/p>\n<\/td>\n<\/tr>\n

\n

MOPE-500<\/strong><\/p>\n<\/td>\n

\n

58.23\u00b13.20<\/p>\n<\/td>\n

\n

59.13\u00b15.24<\/p>\n<\/td>\n

\n

.062<\/p>\n<\/td>\n

\n

37.81\u00b16.64<\/p>\n<\/td>\n

\n

38.82\u00b14.49<\/p>\n<\/td>\n

\n

.059<\/p>\n<\/td>\n<\/tr>\n

\n

MOPE-1000<\/strong><\/p>\n<\/td>\n

\n

57.29\u00b12.96<\/p>\n<\/td>\n

\n

58.52\u00b15.24<\/p>\n<\/td>\n

\n

.061<\/p>\n<\/td>\n

\n

35.44\u00b14.72<\/p>\n<\/td>\n

\n

35.71\u00b14.49<\/p>\n<\/td>\n

\n

.051<\/p>\n<\/td>\n<\/tr>\n

\n

MOLE-500<\/strong><\/p>\n<\/td>\n

\n

59.25\u00b11.88<\/p>\n<\/td>\n

\n

60.72\u00b16.05<\/p>\n<\/td>\n

\n

.050*<\/p>\n<\/td>\n

\n

36.25\u00b12.50<\/p>\n<\/td>\n

\n

37.25\u00b16.18<\/p>\n<\/td>\n

\n

.052<\/p>\n<\/td>\n<\/tr>\n

\n

MOLE-1000<\/strong><\/p>\n<\/td>\n

\n

58.43\u00b12.20<\/p>\n<\/td>\n

\n

59.60\u00b15.41<\/p>\n<\/td>\n

\n

.063<\/p>\n<\/td>\n

\n

35.80\u00b12.38<\/p>\n<\/td>\n

\n

35.57\u00b15.35<\/p>\n<\/td>\n

\n

.049*<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

*<\/strong> P value <0.05 indicating Statistical significance<\/p>\n\n\n

Table 6: Renal Function Tests: BUN, Serum creatinine<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
 <\/td>\n\n

BUN mg\/dl<\/strong><\/p>\n<\/td>\n

\n

Creatinine mg\/dl<\/strong><\/p>\n<\/td>\n<\/tr>\n

 <\/td>\n\n

Baseline<\/strong><\/p>\n<\/td>\n

\n

Week 7<\/strong><\/p>\n<\/td>\n

\n

P value<\/strong><\/p>\n<\/td>\n

\n

Baseline<\/strong><\/p>\n<\/td>\n

\n

Week 7<\/strong><\/p>\n<\/td>\n

\n

P value<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Control <\/strong><\/p>\n<\/td>\n

\n

15.04\u00b1.05<\/p>\n<\/td>\n

\n

18.35\u00b1.03<\/p>\n<\/td>\n

\n

.000*<\/p>\n<\/td>\n

\n

.60\u00b1.02<\/p>\n<\/td>\n

\n

.93\u00b1.03<\/p>\n<\/td>\n

\n

.000*<\/p>\n<\/td>\n<\/tr>\n

\n

Standard<\/strong><\/p>\n<\/td>\n

\n

15.01\u00b1.60<\/p>\n<\/td>\n

\n

15.10\u00b1.14<\/p>\n<\/td>\n

\n

.251<\/p>\n<\/td>\n

\n

.62\u00b1.03<\/p>\n<\/td>\n

\n

.64\u00b1.04<\/p>\n<\/td>\n

\n

.085<\/p>\n<\/td>\n<\/tr>\n

\n

MOPE-500<\/strong><\/p>\n<\/td>\n

\n

16.51\u00b1.73<\/p>\n<\/td>\n

\n

16.76\u00b1.01<\/p>\n<\/td>\n

\n

.167<\/p>\n<\/td>\n

\n

.61\u00b1.02<\/p>\n<\/td>\n

\n

.62\u00b1.02<\/p>\n<\/td>\n

\n

.293<\/p>\n<\/td>\n<\/tr>\n

\n

MOPE-1000<\/strong><\/p>\n<\/td>\n

\n

15.66\u00b1.49<\/p>\n<\/td>\n

\n

15.76\u00b1.35<\/p>\n<\/td>\n

\n

.272<\/p>\n<\/td>\n

\n

.65\u00b1.05<\/p>\n<\/td>\n

\n

.67\u00b1.03<\/p>\n<\/td>\n

\n

.138<\/p>\n<\/td>\n<\/tr>\n

\n

MOLE-500<\/strong><\/p>\n<\/td>\n

\n

16.06\u00b1.11<\/p>\n<\/td>\n

\n

16.17\u00b1.12<\/p>\n<\/td>\n

\n

.136<\/p>\n<\/td>\n

\n

.64\u00b1.03<\/p>\n<\/td>\n

\n

.67\u00b1.06<\/p>\n<\/td>\n

\n

.037*<\/p>\n<\/td>\n<\/tr>\n

\n

MOLE-1000<\/strong><\/p>\n<\/td>\n

\n

15.21\u00b1.03<\/p>\n<\/td>\n

\n

15.30\u00b1.14<\/p>\n<\/td>\n

\n

.121<\/p>\n<\/td>\n

\n

.67\u00b1.06<\/p>\n<\/td>\n

\n

.66\u00b1.05<\/p>\n<\/td>\n

\n

.257<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

*<\/strong> P value <0.05 indicating Statistical significance<\/p>\n\n\n

Histopathological examination<\/strong><\/p>\n\n\n\n

The skin section of the control group showed\natypical squamous cells with keratin pearls on histopathological examination at\nthe end of the study. There was also evidence of hyperkeratosis and chronic\ngranulomatous tissues. The standard (5-FU) treated group exhibited signs of\nhealing with significant scarring at the dermis. MOPE treated group showed\ninfiltration of tissues with fibrosis signifying the process of healing from\nthe inflammation. The MOLE treated group also showed fibrosis and signs of\nhealing in skin tissues. The section taken from the kidney and liver from the\ncontrol group showed marked congestion owing to the effect of DMBA. The\nsections of control kidney showed thickened glomerular basement membrane. The control\nhepatic cells exhibited hydropic degeneration associated with congestion. The\nstandard and test groups had normal kidney and liver morphology in\nhistopathological analysis. Based on the above findings it is suggestive that\nthe carcinoma induced have produced marked changes in the skin, kidney and\nliver of mice in control group while the changes in the standard and test\ngroups were less pronounced and showed signs suggestive of healing.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 5: Control – Skin with tumour showing atypical squamous cells with hyper keratosis.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 6: Standard – Skin with tumour showing dermis scarring.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 7: MOPE 500mg – Skin with tumour showing atypical squamous cells with minimal scarring.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 8: MOPE 1000mg \u2013Skin with tumour showing scarring of dermis.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 9: MOLE 500mg – Skin with tumour showing  fibrosis of dermis with minimal infiltration.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 10: MOLE 1000mg – Skin with tumour showing scarring of dermis.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 11: Control – Liver Tissue showing hydropic degeneration.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 12: Control -Kidney tissue showing inflammatory infiltrates.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
\"\"<\/td>\n\n

Figure 13: Site of tumour after treatment for each group.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

The invitro <\/em>antitumor effects of Moringa\noleifera<\/em> leaf extract by Methyl Thiazolyl Tetrazolium bromide (MTT) assay\nhave already been studied. It was observed that chloroform fraction showed\nhigher anticancer activity. The fractions exhibited a dose dependent inhibition\nof cancer cells with maximum activity at 416 \u00b5g mL-1.11<\/sup><\/p>\n\n\n\n

It has earlier been observed that the\npolyphenolic fraction of Moringa Oleifera<\/em> leaves had potent antioxidant\neffect and demonstrated the ability to scavenge free radicals, metal chelation\nproperties and offered protection against DNA nicking. It was also evident that\nleaf extracts could prevent oxidative stress in rat tissues.12<\/sup><\/p>\n\n\n\n

The anticancer effects of Moringa Oleifera<\/em>\nleaf and bark extracts in cancer cell lines of breast and colon have been\nexplored and the extracts have found to have anticancer potential.13<\/sup><\/p>\n\n\n\n

The anti-tumour effect of Moringa oleifera<\/em>\nleaf extract in immunodeficient nude mice has been assessed against human\nhepatocellular carcinoma (Hep G2) cells using a hollow fibre assay. Hollow\nfibres containing HepG2 cells were implanted subcutaneously in mice and leaf\nextract was given orally for four days. The recovered fibres were analyzed by\ntrypan blue exclusion assay and it was observed that HepG2 cells decreased by\n60% at a dose of 200mg\/kg of moringa leaf extract.14<\/sup><\/p>\n\n\n\n

The cytotoxic effect of ethyl acetate leaf\nextract in lymphoma ascites model of Dalton has been evaluated. Lymphoma was\ninduced in mice by intraperitoneal injection of DLA (Dalton\u2019s Lymphoma ascites)\nand moringa ethyl acetate extract treatment given orally for ten days. It was\nobserved that the treated mice had significantly longer survival rate when\ncompared with the untreated mice.15<\/sup><\/p>\n\n\n\n

The active substances such as phenolic acid,\nflavonoids and niazimicin in M.oleifera<\/em> prevent the progress of cancer\nby down regulation of genes controlling tumour progression and induction of\napoptosis.5<\/sup><\/p>\n\n\n\n

Moringa oleifera <\/em>flower is found to have trypsin inhibitors that has shown potent antitumor and antiangiogenic effects on sarcoma cells.16 <\/sup>Probably this could have been the cause of antitumor effect of Moringa oleifera<\/em> pod and leaf, though in the current study the mechanisms of anti-tumour effect were not studied.<\/p>\n\n\n\n

It is evident from our study that ethanolic\nextracts of Moringa Oleifera<\/em> pod and leaf have anticancer effect against\n7,12-DMBA induced skin cancer in mice. The tumour size reduced to zero, the\nbody weight showed an improvement after the initial fall due to cancer\ninduction. The haematological and biochemical parameters exhibited a response\nsimilar to the standard drug when treated with the pod and leaf extracts of Moringa\nOleifera<\/em>. The histopathological examination of the skin tissue after\ntreatment revealed that the pod and leaf extracts had shown a complete tumour\nregression. <\/p>\n\n\n\n

Moringa Oleifera<\/em> pod and leaf extracts have shown reduction in\nskin tumour size and improvement in haematological, biochemical parameters and\nhistopathological examination in the current study.<\/p>\n\n\n\n

Conclusion<\/strong><\/p>\n\n\n\n

Skin carcinoma is one of the most common\ntumours in humans. Our study exhibits a dose dependent anticancer effect of Moringa\noleifera<\/em> pod and leaf extracts in mice which were comparable with the\nstandard drug 5-Fluorouracil. Hence Moringa Oleifera<\/em> pod and leaf can be\nconsidered as an adjuvant therapy for skin cancers along with the standard\ntreatment if further studies are conducted on human subjects.  <\/p>\n\n\n\n

Acknowledgement<\/strong><\/p>\n\n\n\n

All the authors are thankful to\nChettinad Academy of Research and Education for providing the facilities for\nthe conduct of this research.<\/p>\n\n\n\n

Conflict of Interest<\/strong><\/p>\n\n\n\n

The authors declare no conflict of interest.<\/p>\n\n\n\n

Funding Sources<\/strong><\/p>\n\n\n\n

 This\nresearch was not funded by any external agency.<\/p>\n\n\n\n

References<\/strong><\/p>\n\n\n\n

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