This report summarized all the result of this project, especially in the investigation of the immunomodulating effect of Gynura divaricata, which I hope can give more knowledge to future researchers who have interest to open further research about Gynura divaricate. I also want to thank Sir Fandi Sutanto, S.Farm., M.Si for his help during this internship period and also Miss Putri who is always ready to prepare our needs. Effect of Gynura divaricata extract compared to control on phagocytic activity through carbon scavenging test.
The immunomodulatory effect can be defined as changes in the immune response to a normal level. Traditional herbal medicine is receiving a lot of attention in the field of human health these days, one of the reasons being that they are believed to have an immunomodulatory effect. Gynura divaricata is a popular Asian herb widely used as an alternative medicine for its bioactive compounds such as phenolic acid and flavonoid.
These compounds are believed to have an immunomodulatory effect in previous research, while no previous research has been done to verify the potential immunomodulatory activity of Gynura divaricata. Therefore, the aim of this study is to investigate the potential immunomodulatory activity of Gynura divaricata using a carbon clearance assay. But interestingly, the immunomodulatory activity can be observed by calculating the ratio of liver to spleen and body weight.
We found that there was an immunomodulatory activity, as shown by the proliferation of macrophages and lymphocytes (B and T cells) in the liver and spleen, respectively.
INTRODUCTION
Host Institution/Company
- Description about the company
- Description of Department
- Product of the Host Institution/Company
Management, International Business Management in Creative and Digital Marketing, and International Business Management in International Applied Accounting. This institute publishes a journal in the field of life sciences called Indonesian Journal of Life Science. I3L also carries out community service activities to share knowledge about science as a contribution to society.
I3L is also active in implementing a blood donation program which aimed to contribute to the need for blood in the health sector.
PROJECT DESCRIPTION
Internship Project
- Project Background
- Scope of the project
- Objective / aims
- Problem Formulation and Proposed Solutions
Thus, further research needs to be conducted to investigate the potential immunomodulatory effect of Gynura divaricata. Characterization of the extract was also carried out to check the concentration of flavonoid and phenolic content. To study and investigate the potential immunomodulatory activity of Gynura divaricata using carbon clearance assay.
Immunomodulatory aims to bring the body's immune response back to normal levels by using both the innate and adaptive arms of the immune response. Dried leaves of Gynura Divaricata were weighed and then ground into powder using a crusher, followed by reflux extraction with 60% ethanol to extract the powder into liquid form. The aqueous extract of the leaves was then filtered using Oil-less Piston Vacuum Pump Filtration.
The samples were prepared with 30 mg of the crude extract and then diluted with the mixture of methanol and water (2:8). The concentration of the flavonoid content was determined and the value was expressed in the form of mgQE/g. The phenolic content was measured with Folin-Ciocalteu Assay with gallic acid as the standard since gallic acid is one of the natural and stable phenols, also quite cheap compared to others.
Samples were prepared using 100 mg of crude extract, then diluted with a mixture of methanol and water (1:4). The content of phenols was then calculated and the value was expressed in the form of mgGAE/g. Group C as Stimuno 24 was treated with a dose of Stimuno forte 8.8 mg/kg body weight every 24 hours.
Group D as extract 12 was given Gynura divaricata extract at a dose of 8.8 mg/kg body weight every 12 hours. Finally, group E as a 24-hour extract was given Gynura divaricata extract at a dose of 8.8 mg/kg body weight every 24 hours. The mice were sacrificed, the liver and spleen were removed and weighed, then the weight and appearance of the organs were compared.
FINDINGS
RESULT
- Extraction and Characterization of Gynura divaricata
- Survival Rate
- Carbon Clearance Assay
Based on statistical analysis, the survival rate of the stimulus and extract groups increased significantly (p<0.0001) (Figure 1a). While the survival rate of Stimuno 24 hours and extract 24 hours was the same as the control group. Based on statistical analysis, the survival rate of the untreated group was significantly increased compared to the control group, while the survival rate of the 12-hour stimulant and 12-hour extract was significantly decreased compared to the control group (Figure 1b).
Although the 12-hour stimulation group has the highest phagocytic index, but the standard deviation was high. Therefore, no significant difference was observed between the control group and all treatment groups for group I (Figure 2a). While for group II, the control group has the highest survival rate compared to the other groups, but the standard deviation was also high, so no significant difference was observed between the control group and the other treatment groups (Figure 2b ).
According to statistical analysis, there was no significant difference in liver to body weight ratio between control and treatment groups for both batch I and batch II. The ratio of spleen to body weight was also analyzed between groups for both batch I and II. Negative control group has the lowest ratio of spleen to body weight (0.85%) compared to the other groups and extract 12 h has the highest ratio (1.52%) for batch I.
According to statistical analysis, the survival rate of stimulo 12 hours was significantly increased compared to control groups (Figure 4b).
DISCUSSION
Based on the result for batch I, which is now referred to as the diseased population, negative control group has the lowest survival rate because there were few mice died that may be caused by the use of sodium carboxymethylcellulose (CMC-Na) that can promote inflammation. Another possible reason is also due to the presence of hydrocephalus observed in few mice, which is a brain disease caused by the accumulation of cerebrospinal fluid in the brain cavity (Zou et al, 2020). Meanwhile, the survival rate of stimulo and extract groups was higher compared to negative control groups, this may be due to the immunostimulatory effect of both stimulo and Gynura divaricata.
Based on the result for group II, which is now referred to as the healthy population, the condition of the mice was much better compared to group I, as seen in the high survival rate of the untreated group and the high survival rate of the of control, which was comparable to stimuno. and bring out groups. The survival rate of all groups in group II was better compared to group I, this may be due to the absence of hydrocephalus in group II. It can also be suggested that the Gynura divaricata extract works in diseased mice by stimulating the immune response, but it does not really affect healthy mice, therefore, no significant effect is observed in group II, as seen in the survival rate.
Therefore, phagocytic index of mice treated with Gynura divaricata extract should be higher compared to control, or can be similar or more compared to stimulo group. Although on the result, for both group I and group II, no significant difference was observed between the treatment and negative control groups, this may be caused by several factors, including the carbon that did not reach the bloodstream when the blood was collected . . Despite the fact that the liver is not categorized as a primary or secondary lymphoid organ, it plays a role in the immune response due to the presence of resident macrophages.
Macrophages are crucial to the body's defense mechanism at every level, both innate and acquired immunity. In batch I, the liver to body weight ratio of stimuno 12 h, stimuno 24 h, extract 12 h and extract 24 h showed an increase although there was no significant difference compared to the negative control group. For batch II, all other treatment groups also had a higher liver to body ratio compared to the control group, although there was no significant difference based on the statistical analysis.
No significant difference in spleen-to-body ratio was observed in series I, although spleen weight was greater in the treated groups compared to the control. While for Series II, the spleen-to-body weight ratio at the 12-hour stimulus was significantly different compared to the control groups, indicating that there was lymphocyte proliferation in the spleen. The fact that the flavonoid in the extract could increase the number of lymphocytes and stimulate spleen proliferation suggested that the general process of lymphocyte cell proliferation occurred when antigens bound to the surface of T and B cells were combined with IL-1 from APCs.
CONCLUSION AND RECOMMENDATION
SELF REFLECTION
The result of one-way ANOVA test against the significant difference in survival rate of mice Batch I Dunnet's Multiple Comparison Test Mean Difference Significant Summary P-value. The result of one-way ANOVA test against the significant difference in survival rate of mice Batch II Dunnett's multiple comparison test Mean difference Significant summary P-value. The result of one-way ANOVA test against the significant difference in phagocytic index of mice Batch I Dunnet's multiple comparison test Mean difference Significant summary P-value.
The result of ANOVA one-way test after the significant difference in phagocytic index of mice Batch II Dunnet's multiple comparison test Mean difference Significant Summary P-value. The result of ANOVA one-way test to the significant difference in liver index of mice Batch I Dunnet's multiple comparison test Mean difference Significant Summary P-value. The result of ANOVA one-way test to the significant difference in liver index of mice Batch II Dunnet's multiple comparison test Mean difference Significant Summary P-value.
The result of one-way ANOVA test against the significant difference in spleen index of mice Batch I Dunnett's multiple comparison test Mean difference Significant summary P-value. The result of one-way ANOVA test against the significant difference in mouse spleen index Batch II Dunnett's multiple comparison test Mean difference Significant summary P-value. IMMUNOTIMULATORY EFFECT OF METHANOL EXTRACT OF FLAMBOYANTE LEAF [Delonix regia (Boj. ex Hook.) Raf.] IN MICE.
BIOACTIVE COMPONENTS OF GYNURA DIVARICATA AND THEIR POTENTIAL USES IN HEALTH, FOOD AND MEDICINE: A MINI-REVIEW.
