A prospective, open-label, three arm, single center, controlled study to assess the immune-boosting activity of Mulmina® Mango and Mulmina® Amla Orange health drink in asymptomatic, mildly symptomatic COVID-19 patients and healthcare workers posted to COVID wards

Background 

The immune system is the protective mechanism in a living organism to protect it against harmful pathogens like bacteria, fungi, viruses, harmful substances, germs, and environmental pollutants and helps in maintaining the health and homeostasis of an organism¹.

Since December 2019, the coronavirus SARS-CoV-2 (COVID-19) pandemic has created havoc throughout the world. The mutating nature of the virus has thrown a challenge to the entire scientific and medical field and made it difficult to develop an exact cure for this viral outbreak. Scientists worldwide are consistently trying out to develop an effective treatment, whether synthetic or natural. Recent trends in developing newer treatments include the new meta medicine, which focuses on curing the disease by considering it as the whole body. Hence, the body’s immunity should be strong. Nature is always healing, and so are the natural products and thus play an important role in maintaining and enhancing the natural immunity of every individual. In this context, natural immunity boosters have also gained interest throughout the world.

Due to the COVID-19 pandemic situation, different synthetic medicines, herbal medicines, traditional Chinese medicines, and natural moieties are being studied, and clinical studies are performed ². India is since long is facing the burden of undernutrition and immunity deficiency, mostly observed in the children who are consuming less than 80% of RDA (Recommended Daily Allowance) of vital vitamins like A, B, C, D, and E, minerals like calcium, magnesium, zinc, selenium, iron, iodine, and are more susceptible to the risk of developing different infections leading to poor health and overall retarded growth. Additionally, the consumption of food with less antioxidant value is acting as a catalyst for the development of most diseases. Hence, to enhance the body’s natural immunity using different supplements, health drinks containing the appropriate amount of vitamins, minerals, fibres, micronutrients, and antioxidants have become necessary ³. 

Due to the increased demand of the common public, the use of natural immunity boosters, health supplements, health drinks, energy drinks are gaining attention worldwide and are being studied. Various in vivo, in vitro, and clinical studies are being evaluated to screen the best effective treatment .and improving the overall immunity. Some clinical studies are discussed below.

Study design and Method

To better understand the effect of health drinks, 120 patients who were asymptomatic or mildly symptomatic were included in the study and divided into three groups. 

Group І (n=40): Patients received health drink A, twice a day (Treatment A)

Group II (n=40): Patients received health drink B twice a day (Treatment B)

Group III (n=40): Standard Treatment C).

The safety and efficacy of the health drinks were evaluated on 0 days (before the start of heath drink), 14th day, and 28th day of using the health drinks. The blood samples were collected from the three groups, and plasma analysis was done using ELISA kits for serum immune biomarkers and serum antioxidant markers estimations.

Results

Some of the parameters evaluated for serum immune biomarkers included serum amyloid A (SAA), Cytokine interleukin 4(IL-4), Cytokine interleukin 6 (IL-6), Interferon-gamma (IFN-γ), and C- reactive protein (CRP). Serum antioxidant markers included Antibody immunoglobulin A (IgA), Superoxide dismutase (SOD), and catalase (CAT): Glutathione (GSH) count, Malondialdehyde (MDA) count.

All the enrolled patients had increased SAA. A statistically significant increase (p<0.05) was observed between group A vs. C and betweengroup B vs. C. There was no statistically significant difference observed between groups A vs. B indicating no change in cytokine values from day 0 till day 28 in patients being treated with the health drinks. 

Levels of cytokine interleukin 4 (IL-4) showed no significant increase from day 0 to day 28 in both groups treated with health drinks. Levels of cytokine IL-6 were more on day 14, but the health drinks were effective in restoring them to baseline level by day 28. A mild reduction was observed in IFN-γ values in asymptomatic and mild symptomatic COVID-19 patients having health drink A. A sharp reduction in IFN-γ values was observed in mild asymptomatic patients by day 28. Inflammatory marker CRP values were reduced by day 14 as compared to the control group. This reduction was further reduced in mildly symptomatic patients in both the groups having the health drink till the 28th day. Antibody immunoglobulin A (IgA) increased in the three sub-groups being treated with the health drinks by the 28th day.

A significant increase was observed in SOD, CAT, and GSH values by day 28 with both the health drinks as compared with the control group with standard treatment in asymptomatic and mildly symptomatic patients. Malondialdehyde (MDA) value reduced in all the groups having the health drink.

Discussion

A higher IL‐6/IFN‐γ ratio relates to an enhanced cytokine storm favoring lung damage. The study showed a significant reduction in the IL‐6/IFN‐γ ratio in all patients proving their potential role in controlling the dysregulated or imbalanced immune system. Further, the reduction in the values OF C- reactive protein (CRP), suggesting that health drinks had excellent anti-inflammatory activity.

Oxidative stress is an imbalance between the production of reactive oxygen species (ROS) and the elimination of ROS from the body (antioxidant property) to maintain the body’s homeostasis and redox balance. Viral infection can affect and disturb this homeostasis by increasing ROS production, thus creating oxidative stress, which triggers an increase of redox signaling, activation of transcription factors, stimulation activity of pro-inflammatory and pro-fibrotic cascades DNA damage, and also stress-dependent kinases induction. Also, increased ROS production causes a destructive effect on the surrounding host cells of any organs, leading to cell damage. In the case of SARS-CoV-2-infected patients, the lung cells get the most affected ³. Hence, reduction in Malondialdehyde (MDA) value and a significant increase in the antibody immunoglobulin A (IgA), SOD, CAT, and GSH values indicate the effectiveness of the health drinks as an antioxidant. 

Conclusion

The discussed clinical trial study provides reliable and strong evidence that natural components of the health drinks exhibited strong antioxidant and immune-boosting properties in mildly symptomatic and asymptomatic COVID-19 patients, establishing that natural ingredients have the potential to promote better immunity to humans.

References:

1. Jayanthi CR. A prospective, open-label, three-arm, single-center, controlled study to assess the immune-boosting activity of Mulmina mango and mulmina amla orange health drink in asymptomatic, mildly symptomatic COVID 19 patients and health care workers posted to COVID wards.  Clinical trials registry India. CTRl/2020/06/025702 [Registered on: 08/06/2020]. Accessed on 24/06/2021 from http://www.ctri.nic.in/Clinicaltrials/pdf_generate.php?trialid=44376&EncHid=&modid=&compid=%27,%2744376det%27
2. Mohammad Asaduzzaman Chowdhury; Nayem Hossain , M. A. K., Md. Abdus Shahid , Ashraful Alam. Immune response in COVID-19: A review. Journal of Infection and Public Health, 2020, 13, 1619-1629.
3. Marina Darenskaya 1, L. K., Sergei Kolesnikov 1. The Association of Respiratory Viruses with Oxidative Stress and Antioxidants. Implications for the COVID-19 Pandemic. Curr Pharm Des 2021, 27, 1618-1627.
4. Evgenii M., E. I., Andrey V. Grechko, Kozlovand Alexander N. Orekhov,Wei-Kai Wu,Antonina V. Starodubova. Involvement of Oxidative Stress and the Innate Immune System in SARS-CoV-2 Infection. Diseases 2021, 9, 1-15.