INTRODUCTION:

Medicinal plant must stay used as a lesions curative cure for centuries popular virtually and in the conduct of a number of diseases separately from lesions healing. These include diseases associated to the membranes, respirational tissues, peptic organs, imitation, the circulation, against tumor and in contradiction of infections¹. Plantago, a class in the family Plantaginaceae (order Lamiales) with around 260 species. The little plants typically must a heavy tassel of basal leaves and lengthy, leafless debris bearing a terminal spike of little blossoms². Plantago lanceolata historical underpinnings, Plantago gets from the Latin word planta, signifying “young plant” or “sole of the foot”. The seeds of this plant have laxative properties because of the presence of polysaccharides in it. The leaves of Plantago lanceolata have a bitter flavor and are astringent, demulcent, mildly expectorant and ophthalmic³.

The roots of Plantago lanceolata are used as a remedy for rattlesnakes, insect stings and bites. Plantago lanceolata looks near be actual multipurpose totaling to the medication breakfront existence antimicrobial and antioxiidant^4,5. The greeneries correspondingly are used to brand a drink those performances as an active coughing medication⁶. It can likewise diminish the dangers of metabolic conditions by controlling glucose levels in diabetic patients, lessening lipid levels in hyperlipidemics, and supporting weight control⁷. The plant is additionally used to treat issues of the stomach related framework, for example, constipation, dysentery “infection of the intestines subsequent in severe diarrhea with the presence of blood and mucus in the feces”, the husk of plant is utilized to treat irritable bowel syndrome and genitourinary tract problems. Also husk, seed and mucilage can be used super disintegrants^8,9.

MATERIAL AND METHODS:

1- Plantago lanceolata:

Leaves of Plantago lanceolata were supplied from the local Iraqi markets during September/2020 and recognized by Dr. Ibrahim S. Al-Jubouri, College of Pharmacy, Mustansiriyah University, Iraq.

2- Plant Extract:

Aqueous extract of Plantago lanceolata was prepared according to¹⁰ with a weight of 10g of herb, it was placed in a clean flask and 250ml of distilled water supplementary to it, and then it remained located in the shaking incubator at a temperature of 28C° for a period of 30 minutes, then filtered using medical gauze, and then the filtrate was distributed on plates. Dried the water in the oven at 37C° until the water evaporated completely to obtain the dry powder for the aqueous extract. Each sample was placed in sealed tubes, and placed in refrigerator at 4C° until use.

3- Dose of plant:

In albino male mice. A dose of 400mg/kg was tested depending on LD50 of Plantago lanceolata to 2,940 mg/kg¹¹.

4- Research laboratory Animals:

Albino male mice (Mus muscles) were the laboratory animals which supplied from the Biotechnology Research Centre (Al-Nahrain University). Their ages at the first of research were eight-ten weeks, and their weight was twenty three –twenty seven grams.

5- Experimental Design:

The experimental designs were divided into three groups, as shown below, in which each group contain 4 animals

· Group I: Mice were managed with Plantago lanceolata aqueous extract at dose of (400mg/kg).

· Group II: Mice were administrated with the MTX (7th days) at dose of (40mg/kg).

· Group III: Mice were administrated with the MTX (1st and 2ed days) + Plantago lanceolata aqueous extract (from 3rd to 7th day)

The mice were injected intraperitoneally (IP) as a single dose of plant extract (0.1ml) per a day for 7 days. In day 8, the mice sacrificed. Over all,12 mice were used.

6- Tissue Preparation for Histology Preparation:

The organs (testis and spleen) per mouse are set for histopathologically learning by way of termed¹². Sections remained stable in (10%) formalin about 24h, afterward dehydration through measured series of alcoholic liquid ranged from (30 to 100) % for (5) min apiece. Formerly the tasters cleared in two changes of xylene beforehand fixed in paraffin wax for sectioning. Cross sections of (5)µm fatness are ready and stained with hematoxylin (Harison) and eosin rendering to typical technique. Any variations in sections are detected by light microscope.

RESULT:

In mice treated with plant extract, the section of the testis showing normal developed and maturation of spermatogoni cells inside seminephrous tubules with presence sperms indie the lumen.

Fig (1) (testis): section of the testis showing normal developed and maturation of spermatogoni cells inside seminephrous tubules with presence sperms indie the lumen. (H & E) (X40).

While the results of spleen tissue represented widening of white pulp and reduction of red pulp

Fig (2) (spleen) spleen tissue represented widening of white pulp and reduction of red pulp. (H & E) (X40).

While the results of interaction group for (testis and spleen) showing the ability of plant extract to counter act the damaged induced by methotrexate drug.

Fig (3) (testis): Section showing normal development and maturation of spermatogoni cells of seminephrous tubules with presence of sperms inside the lumen. (H & E) (X40).

Fig (4) (Spleen): Section showing widening of white pulp and reduction of red pulp with formation of germinal center. (H & E) (X40).

The results of mice treated with methotrexate indicated the ability of drug to cause damage to all organs tested

Fig (5) (testis): The destructive variations give the impression additional obviously that mains to nonappearance of spermatogenesis through tinny of the wall of seminiferous tube of the testes. (400X; H and E)

Fig (6) (spleen): section showing necrosis of splenic cells. (400X; H and E)

DISCUSSION:

Today, all the scientific are searching about ideal drug without un desirable effect for that reason uses of medical plants have been increasing highly because it contain numerous phytochemical compounds such as tannins, flavonoids, caffeic acids,etc that have been showed high potentially effect against harmful factors or pathogensis of disease without side effect^13,14 Morever, they are free toxic effect and present suitable folklore medicine¹⁵.

Plantain (Plantago lanceolata) is traditionally used for the treatment of the common cold and related symptoms such as cough¹⁶. Plantain is one of the perpetual herbs contain many bioactive compounds like caffeic acid derivatives (in particular acteoside), iridoid glycosides (are aucubin, catapol)¹⁷, which have anti-oxidative activity. Plantago lanceolata dry extract is a composite mixture of phytochemical ingredients¹⁸. Huge studies have been agreed in regard to the antioxidant properties of plantago, and in general their findings are in agreement with the present results where plantago leaves and seeds contain carbohydrates, lipids, alkaloids, caffeic acid derivatives, flavonoids, iridoid glycosides, vitamins and other organic substances remaining to its diverse medicinal properties. Furthermore, tannins exhibited antibacterial activity by many previous studies ^19,20. Some studies refer that tannins and flavonoid and phenol are obtain in aqueous extract of plant²¹, also, treatment with aqueous extract of Planago ovata in root of meristem cells of Vicia faba has been appeared mitotic -inhibition and lead to many chromosomal aberrations²².

Balance between the antioxidants and free radicals is playing a vital role for good health. The risk possibility of free radicals has been increasing gradually in our life. Many disease such as cancer, atherosclerosis inflammatory joint disease are a result of imbalance between the antioxidants and the free radicals^23,24. In addition Each of the constituents has unique medicinal property²⁵ and these compounds have activity as free radicals scavenger. Such belongings consider to be attained over numerous metabolic paths; for occurrences, prevent the establishment of free radicals, overturn sequence beginning and contravention sequence proliferation reactions²⁶, cumulative the action of antioxidant enzymes for instance superoxide dismutase and glutathione transferase and de novo anti-oxidant²⁷ and acclimation wherever the indication for the manufacture and scavenge free radicals formation and transfer it the anti-oxidant to the right sight²⁸. The flavonoid compounds assist for bio-functions, with special reference to anti-mutagenic activity²⁹. P. L. was grown up hydroponically in a water medium and stimulation by different concentration lead ion on different times and temperature regimes to evaluated lead detoxification effective by different parts of the plant (stems, leaves, roots and whole plant). Previous study declared that roots of the plant presented the highest elimination rate of lead than additional parts³⁰. All of above clarified that the protective role of plantago lanceolata is due to its antioxidant and scavenger capability of the bioactive compound.

ACKNOWLEDGEMENT:

The authors has been appreciated to Mustansiriyah University (www.uomustansiriyah.edu.iq) and Al-Nahrain University Baghdad, Iraq to their helpful to complete this research.

REFERENCES:

1. Al-Ezzy, R.M., Abood, K.W., Abdullah, J.M. and Rdeef, A.K. Evaluation of Sperm Head Abnormality Traits and Anti-bacterial Properties of Achillea millefolium Methanolic Extract on Methotrexate- induced Albino Male Mice IJDDT.2019, 9 (3).

2. Hammami S., Debbabi, H., Jlassi, I., Joshi, R. K., and El Mokni, R. Chemical composition and antimicrobial activity of essential oil from the aerial parts of Plantago afra L.(Plantaginaceae) growing wild in Tunisia. South African Journal of Botany, 2020 132: 410-414. DOI:10.1016/j.sajb.2020.05.012

3. Abajy, M. Y., Nayal, R., and Takla, S. Investigation of the Effect of Potentilla erecta and Plantago lanceolata on Bleeding Time in Mice. Res. J of Aleppo univ, Medicine Sience Series. 2015

4. Bahadori, M. B., Sarikurkcu, C., Kocak, M. S., Calapoglu, M., Uren, M. C., and Ceylan, O. Plantago lanceolata as a source of health-beneficial phytochemicals: Phenolics profile and antioxidant capacity. Food Bioscience.2020, 34: 100536. doi.org/10.1016/j.fbio.2020.100536

5. Oliva, A. I., Monleon, G. A., Rada, F. D. J. D., Oteiza, L. L., Rodriguez, G. S., Garromendiola, G. J. K., and Martin, V. J. A pilot study of efficacy and safety of Plantago lanceolata and Primula veris, in the treatment of the common cold. Archives of Pharmacy Practice.2020, 11(3): 1-7.

6. Khan, F. A., Bhat, S. A., and Narayan, S. Wild edible plants as a food resources: Traditional knowledge. Division of post-harvest technology sher_e_Kashmir 2017 university of Agricultural sciences and technology of Kashmir. India.

7. Díez, R., García, J. J., Diez, M. J., Sierra, M., Sahagun, A. M., and Fernández, N. Influence of Plantago ovata husk (dietary fiber) on the bioavailability and other pharmacokinetic parameters of metformin in diabetic rabbits. Biomedcentral complementary and alternative medicine.2017, 17(1): 298. doi.org/10.1186/s12906-017-1809-x

8. Shailesh Sharma, Sudhir Bharadwaj, G. D. Gupta. Fast Dissolving Tablets of Promethazine Theoclate by Using Natural Superdisintegrants. Research J. Pharm. and Tech. 1(3): 2008; 218-224.

9. Girish kumar, R. K. Khar, Tarun Virmani, Vikash Jogpal, Reshu Virmani. Comparative Evaluation of Fast Dissolving Tablet of Atorvastatin Calcium using Natural and Synthetic Super Disintegrating Agents. Research J. Pharm. and Tech 2018; 11(11): 5001-5007. doi: 10.5958/0974-360X.2018.00912.5

10. Velasco-Lezama, R., Tapia-Aguilar, R., Román-Ramos, R., Vega-Avila, E., and Pérez-Gutiérrez, M. S. Effect of Plantago major on cell proliferation in vitro. Journal of Ethnopharmacology, 2006;,103(1), 36-42. doi.org/10.1016/j.jep.2005.05.050

11. Rios, J. L., Recio, M. C., and Villar, A. Screening methods for natural products with antimicrobial activity: a review of the literature. Journal of Ethnopharmacology, 1988,;23(2-3), 127-149. doi.org/10.1016/0378-8741(88)90001-3

12. Niran A. Ibrahim, Hanady S. Al-Shmgani, Ruqaya MI Al-ezzy. Cytarabine induced reproductive histopathological changes in albino male mice. Journal of Biotechnology Research Center. 2017;11(1):6-12. doi.org/10.24126/jobrc.2017.11.1.489

13. Shanmuga Sundaram C., Subramanian S. Biochemical Evaluation of Hypoglycemic Activity of Musa paradisiaca (Plantain) Flowers in STZ- induced Experimental Diabetes in Rats. Asian J. Research Chem. 2011;4(5):827-833.

14. T. Sheela Rani, V. Gopal, S. Seethalakshmi, K. Chitra. Quantification and Screening of Phytoconstituents in Selected Medicinal Plant. Research J. Pharm. and Tech. 2013; 6(11): 1235-1236. DOI: 10.5958/0974-360X

15. Sirisha, N., et al. A Review on Herbal Diuretics. Research Journal of Pharmacy and Technology, 2011; 4(3): 335-348.‏

16. Abdalah M.E., Al-Ezzy R.M., Okhti Z.A Protective role of Viola odorata against hepatotoxicity induced by methotrexate in albino male mice . J. Pharm. Sci. & Res.2018;10(11): 2775-2785.

17. SILVEIRA, Dâmaris, et al. COVID-19: is there evidence for the use of herbal medicines as adjuvant symptomatic therapy?. Frontiers in Pharmacology, 2020, 11: 1479.‏doi.org/10.3389/fphar.2020.581840

18. Ishiguro, K.; Yamaki, M.;TAKAGI, S. Studies on the iridoid related compounds. I. On the antimicrobial activity of aucubigenin and certain iridoid aglycones. Yakugaku zasshi: Journal of the Pharmaceutical Society of Japan, 1982;102.8: 755-759.‏ DOI: 10.1248/yakushi1947.102.8_755

19. Saradhai, P., et al. Effect of Tannins of Herpestis monneira against Selective UTI Pathogens. Research Journal of Pharmacy and Technology, 2010; 3.3: 832-834.

20. RANI, T. Sheela, et al. Quantification and Screening of Phytoconstituents in Selected Medicinal Plant. Research Journal of Pharmacy and Technology, 2013; 6.11: 1235-1236.‏

21. Krishnamoorthy, B. S., Et Al. Preliminary Studies on Phytochemicals of Solvent Extracts of Eichhornia Crassipes. Research Journal of Pharmacy and Technology, 2014; 7.6: 635-637.‏

22. Yadav, Himshikha; KUMAR, Sushil. An Assessment of Cytotoxic Potentiality of Plantago ovata Husk Aqueous Suspension on the Root Meristem Cells of Vicia faba L. Research Journal of Pharmacy and Technology, 2017; 10.7: 2053-2057.‏ DOI: 10.5958/0974-360X.2017.00358.4

23. Suresh, Preeti K.; SAH, Abhishek K.; DAHARWAL, Sanjay J. Role of free radicals in ocular diseases: An overview. Research Journal of Pharmacy and Technology, 2014, 7.11: 1330-1344.‏

24. Preeti Tiwari. Phenolics and Flavonoids and Antioxidant Potential of Balarishta Prepared by Traditional and Modern Methods. Asian J. Pharm. Ana.2014; 4(1): 05-10.

25. Budzianowska, Anna; Budzianowski, Jaromir. A New Flavonoid, A New Phenylethanoid Glycoside and Related Compounds Isolated From The Inflorescences Of Plantago Lanceolata L. Natural Product Research, 2021, 1-12.‏ DOI: 10.1080/14786419.2021.1888289

26. Ibraheema R.M., Abbood K.W., Ad'hiah A.H Anti-inflammatory Effects of Hypericum triquetrifolium Turra Methanol Extract in Albino Male Mice. Journal of Ethnobiology and Traditional Medicine2015,(125):1039-1045.

27. Al-Ezzy, R.M. Immunological and Cytogenic Effect of Saga (Salivia officinalis) Leaves Extracts on Albino Male Mice and Acute Lymphoid Leukemic Cells. M.Sc. Thesis. Al-Nahrain University /college of science.2007

28. Ali, Akram A.; Al-Homaidan, Ali A. Removal Of Lead Ions From Polluted Water Using Plantago Major L. And Phaseolus Vulgaris L. Aust J Basic Appl Sci, 2007, 1.4: 467.‏

29. Al-Ezzy, R.M., Mhawesh, A,A. and Abbood K.W . Estimation of the whole flavonoid, antioxidant, antibacterial challenge concerning viola odorata (banafsha) methanolic extract. Iraqi Journal of Agricultural Sciences –1027:49(4): 2018566- 551. DOI: https://doi.org/10.36103/ijas.v49i4

30. AlChalabi, R., R. Mohammed, R. Adnan, and S. H. Al-Bayati. The Antibacterial Effect of Ethanolic Extract of Allium Sativum on Biofilm Forming Staphylococcus Aureus Which Cause folliculitis. Int. J. Adv. Res. 5(5): 2017 1175-1182. DOI: 10.21474/IJAR01/4228

Received on 05.10.2021 Modified on 15.12.2021

Research J. Pharm. and Tech. 2022;15(7):3241-3244.

DOI: 10.52711/0974-360X.2022.00543