Antiasthma Activities of Moringa oleifera Lam. Leaves Extract on the Eosinophil Count and Mast Cells in BALB/c Mice

Asthma is a chronic inflammation of the respiratory tract. This study aimed to prove Moringa oleifera Lam. leaves extract's effect on reducing the eosinophil count and mast cells in asthmatic mice. Twenty BALB/c mice that met the inclusion criteria were divided into four groups of standard control (K1), negative control (K2), dexamethasone (K3), and M. oleifera leaves extract (K4). On days 0 and 7, intraperitoneal ovalbumin (OVA) was sensitized. On days 14, 16, and 18, mice were re-sensitized by inhalation using 1% OVA in sterile 0.9% NaCl. On days 18 to 25, K1 and K2 groups were given Na-CMC, the K3 group was given dexamethasone 1.3 µg/day, and the K4 group was given M. oleifera leaves extract 3.9 mg/day. On the 25 th day, the mice were terminated to analyze the eosinophil count and stable bronchiolar mast cells. In conclusion, M. oleifera leaves extract was proven to decrease the eosinophil count with a p-value <0.05 and could stabilize bronchiolar mast cells with a p-value <0.05.


INTRODUCTION
Asthma is a disease with an increasing prevalence rate, and there is a strong interaction between airway epithelial cells and the immune system against the pathogenesis of asthma 1 . The most common immunopathology in asthma is inflammation of type 2 characterized by processes in the airway epithelium involving cytokines 2 . Symptoms of asthma include wheezing, shortness of breath, heavy chest, and coughing at night and early morning. There is muscle contraction in the bronchial walls, swelling of the bronchial mucosa, and increased mucus production in the secretory cells of the bronchial mucosa, resulting in increased resistance to airflow through the narrowed bronchi. This condition occurs in shortness of breath 3, 4 .
The increase in the eosinophil count in asthmatic patients correlated with the severity of the respiratory failure and was predicted to increase the risk of sequential subexacerbations. Immunoglobulin E (IgE) is involved early in the inflammatory cascade and may be considered a cause of allergic asthma 5, 6 .
Moringa oleifera Lam. is considered a miracle plant with many benefits for humans and is credible for providing good nutrition and healing and preventing many

Materials
The materials used in this study were M. oleifera Lam.

Preparation of test animals
The test animals used were female BALB/c mice weighing 20-30 g, which has been acclimatized for a week. All test animals were given drink and standard feed ad libitum. Female BALB/c mice were used because they showed a better response to OVA than males 19 .

Research design
This study used a laboratory experimental quantitative  Significance was defined at the p <0.05 level.

Examination of bronchiolar mast cells
Mast cell examination was carried out by histopathology of the mice's lungs with Toluidine blue staining and observed with a 400x magnification software microscope Image OptiLab Pro 6.1 21 . This variable size scale was a ratio. Data analysis was performed using one-way ANOVA and then analyzed by post hoc test to determine the difference between the two treatment groups. Data were presented as average ± SD. Significance was defined at the p <0.05 level.

Ethical considerations
This research was conducted after obtaining approval from the health research biotech commission of the

Normality and homogeneity analysis of the eosinophil count and mast cells
The data normality test for the eosinophil count was performed with Shapiro-Wilk, with a p-value >0.05, indicating that the data were normally distributed as presented in Table I. As for the homogeneity of the data, the post hoc test results were obtained with a p-value >0.05, indicating that the data obtained for the eosinophil count and mast cells were homogeneous, as shown in Table II.

Examination of eosinophil count
The statistical analysis of the test results for the average, SD, F value, and p-value of the eosinophil count in all groups were presented in Table III

Examination of bronchiolar mast cells
The test results for the average, SD, F value, and p-value of the mast cells in groups K1, K2, K3, and K4 were presented in

ACKNOWLEDGMENT
The authors would like to thank the laboratory technicians and staff of the department of pharmacology, Universitas Islam Sultan Agung, Semarang. We also thank STIKES Cendekia Utama Kudus for the permission for the research. All authors contribute to financing this work.