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Article

Review

J. Microbiol. Biotechnol. 2025; 35():

Published online January 15, 2025 https://doi.org/10.4014/jmb.2408.08028

Copyright © The Korean Society for Microbiology and Biotechnology.

A Systematic Review on the Antimicrobial Activity of Andrographolide

Gayus Sale Dafur1,2, Aiza Harun3, Tuan Noorkorina Tuan Kub1, Ruzilawati Abu Bakar4, and Azian Harun1,5*

1Department of Medical Microbiology and Parasitology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan 16150, Malaysia
2Department of Biology, Federal College of Education Pankshin, Plateau State 933105, Nigeria
3Faculty of Applied Sciences, Universiti Teknologi MARA, Bandar Jengka, Bandar Tun Razak 26400, Pahang
4Department of Pharmacology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan 16150, Malaysia
5Hospital Universiti Sains Malaysia, Jalan Raja Peremppuan Zainab II, Kubang Kerian, Kelantan 16150, Malaysia

Correspondence to:Azian Harun ,       azian@usm.my

Received: August 19, 2024; Revised: October 10, 2024; Accepted: November 5, 2024

Abstract

Andrographolide, a bioactive compound from Andrographis paniculata, has gained attention for its antimicrobial properties, which include antibacterial, antiviral, antifungal, and antiprotozoal effects. As an herbal extract used in traditional medicines, andrographolide also shows promise for developing new antimicrobial agents, especially in the fight against rising antimicrobial resistance. Following the PRISMA 2020 guidelines, 16 peer-reviewed studies published from 2010 to 2024 and focusing on andrographolide’s effects on bacteria, viruses, fungi, and protozoa were reviewed. The quality and bias risk of these studies were assessed using the In Vitro Quality Evaluation Instrument to ensure methodological rigor. The findings demonstrate that andrographolide is effective against bacteria such as Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus. However, its antifungal efficacy is limited, as it was ineffective against Candida albicans and Saccharomyces cerevisiae, but effective against Alternaria solani. It exhibited strong antiviral activity against 2019-nCoV, Dengue virus, and Enterovirus D68, and showed antiprotozoal effects against Plasmodium falciparum and Setaria cervi. Nonetheless, variations in its efficacy across different microorganisms were observed. The quality assessment revealed low bias risk in 11 out of 16 studies (78.57% to 92.86%), while the remaining five had medium bias risk (57.14% to 64.29%), indicating an overall acceptable quality of the studies. Information on andrographolide’s potential and effectiveness across various microorganisms is crucial. Therefore, the purpose of this review was to synthesize the existing data on andrographolide’s antimicrobial activity and assess its potential in combating antimicrobial resistance. This review highlights the need for further research on andrographolide’s antifungal activity, mechanisms of action, clinical safety, toxicity, and potential applications in antimicrobial resistance strategies.

Keywords: Antimicrobial activity, andrographolide, systematic review

Introduction

Medicinal plants are of vast importance to human health [1]. It is well known that herbal extracts can be used to treat a variety of infectious and non-infectious disorders [1, 2]. Novel antimicrobials can be found in plants [3], and those used in traditional medicines include natural and active antimicrobial compounds that are often affordable, safe, and efficient for treating common diseases with microbial origins [4, 5]. According to reports, medicinal plants are the most abundant source of modern medications, traditional medicines, folk medicines, ingredients for synthetic drugs, and pharmaceutical intermediates [6]. New compounds with antimicrobial characteristics are constantly being discovered in plants [7, 8]. Additionally, the discovery and characterization of active components in medicinal plants has tremendously aided in developing innovative medications with potent therapeutic effects to address a range of medical conditions [9]. Numerous plants contain various active ingredients with wide-ranging therapeutic and antimicrobial characteristics. It is therefore crucial to explore these plants and their constituents to develop new antimicrobial drugs, as well as to discover and publicize their therapeutic efficacy.

Andrographolide is a labdane diterpenoid bioactive compound primarily derived from the Andrographis paniculata plant. Due to its diverse biological activities, this plant is extensively utilized in conventional medicine to treat numerous illnesses or infections [10-14]. Andrographolide is shown in numerous studies to have efficacy against a variety of microbiological infections. It exerts antiviral [15-22], antibacterial [23-26], antifungal [27], and antiprotozoal activities [28, 29], making it a viable option for creating new antimicrobial drugs. However, there is little available information and no thorough analysis has been conducted on andrographolide's effectiveness as an antimicrobial agent, including its mode of action or molecular interactions with microorganisms.

In this review, we aimed to assess, synthesize, and summarize the available evidence on the antimicrobial activity of andrographolide against diverse microbes, such as viruses, bacteria, fungi, and protozoa to uncover the current evidence on this topic. Previously, the antimicrobial activity of andrographolide was the subject of a review that evaluated the methodological merits and conclusions of many original research publications and gave an overview of how each of these classes of microorganisms was affected by andrographolide. This review is important because andrographolide has shown promise traditionally in treating various illnesses, as well as in studies performed in vitro against some microorganisms. Unfortunately, the lack of related comprehensive data has prevented determining the spectrum of antimicrobial activity that the compound possesses against microbiological pathogens. In addition, our review offers important information about the effectiveness, spectrum of action, and possible uses of andrographolide against different microbial pathogens, which could help solve a major and pressing problem in public health and microbiology. This information might result in the creation of novel antimicrobial therapies, especially considering the growing concerns about antimicrobial resistance and global health. Finally, this review identifies knowledge gaps that should be filled to direct future studies and enhance both public health and microbiological research.

Characteristics and Advantages of Andrographolide

Andrographolide, a key bioactive compound sourced from Andrographis paniculata plants, serves as a natural alternative to synthetic antimicrobial agents. It offers the advantage of potentially fewer side effects and a reduced risk of promoting resistance. Its long history in traditional medicine underscores its safety. Andrographolide is a colorless, crystalline substance with a distinctly bitter taste and is noted for its diverse biological activities. Its traditional medicinal uses and promising antimicrobial properties, which include antiviral, antibacterial, antifungal, and antiprotozoal effects, make it a strong contender for developing new antimicrobial agents, particularly in combating the growing threat of antimicrobial resistance.

Review Approach and Scope

Design of the Study

The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 guideline [30] was adopted for this systematic review (Fig. 1). PRISMA 2020 is a framework designed to enhance transparency and completeness in systematic reviews by providing a structured checklist and flow diagram for reporting. In this review, PRISMA ensures the clear documentation of study selection, search strategy, risk of bias assessment, and data synthesis, making the review reproducible and comprehensive. Unlike other types of reviews, PRISMA emphasizes standardized reporting, detailed risk of bias assessment, and thorough exploration of heterogeneity among studies. Its application in this study ensures that the review process meets the highest standards of scientific rigor, promoting transparency, replicability, and credibility in the findings.

Figure 1. PRISMA flow diagram of articles screened for inclusion in the systematic review.

Inclusion Criteria

The review included studies that fulfilled the following criteria:

i. In vitro and ex vivo studies evaluating the antimicrobial activity or effect of andrographolide or diterpene lactone against viruses, bacteria, fungi, and protozoa.

ii. Publications with peer review that were published in the English language between January 2010 and January 2024.

iii. Original study findings published in scientific journals.

Exclusion Criteria

The following criteria were used to eliminate studies:

i. Studies that use animals.

ii. Studies not evaluating the antimicrobial activity or effect of andrographolide or diterpene lactone.

iii. Studies not involving bacteria, fungi, viruses, or protozoa.

iv. Studies that used derivatives or analogues of andrographolide.

Search Strategy

Using the terms "antimicrobial" OR "antifungal" OR "antibacterial" OR "antiviral" OR "antiprotozoal" AND "activity" OR "effect" AND "andrographolide" OR "diterpene lactone," we conducted an extensive electronic database search across PubMed, Web of Science, Scopus, Google Scholar, and ScienceDirect to look for articles or studies from January 2010 to January 2024. The search was limited to original, peer-reviewed articles that were written in English. Additionally, we manually reviewed the collected publications' reference lists for any additional pertinent studies.

Search Outcome

In all, 678 papers that discussed andrographolide's antimicrobial properties. A total of 580 peer-reviewed publications were screened after the selection criteria were applied. Then, after duplicates, articles under review, and articles discarded for other reasons were removed, 158 articles were sought for retrieval. Among these, 139 articles were evaluated for eligibility, and 19 articles could not be downloaded since their full texts were not available (Fig. 1). Out of the 139 publications evaluated, 16 peer-reviewed articles met the criteria for inclusion, and thus were included in this review. The title, abstract, and full text of each peer-reviewed paper were independently assessed by all authors to determine the research eligibility. Through group discussion, any discrepancies or conflicts in the authors' eligibility assessment were settled.

Quality Assessment of Included Studies

The quality and bias risk of each included study were assessed using the 12 items or criteria of the In Vitro Quality Evaluation Instrument (QUIN TOOL) of [31], as shown in Table 1. Three reviewers independently evaluated each study, while two reviewers provided confirmation for each assessment. The reviewers discussed any differences or disagreements, and reached a consensus as a group. Each study was evaluated by assigning one of the following scores to every one of the twelve items: 2 for adequately stated, 1 for inadequately stated, 0 for unspecified, and criteria were removed from computation for unapplicable items. To determine the total rating for a specific study, the scores were added together and used to determine the final percentage (%) score, according to the formula: Final percentage (%) score = (overall score / 2 × number of relevant criteria) × 100. Final percentage (%) scores obtained were utilized to categorize each of the studies as low risk of bias (70% and above), medium risk of bias (between 50% and 70%), or high risk of bias (below 50 %).

Table 1 . The score distribution and evaluation parameters for bias risk in the included studies..

S/NCriteria/Check list
Adequately stated (Score = 2)Inadequately stated (Score = 1)Unspecified (Score = 0)Unapplicable (Excluded)
1Clearly defined aims or objectives
2A thorough description of the sample size calculation
3Detailed description of sampling method
4Information about the comparison group
5Detailed description of the methodology
6Operator details
7Randomization
8Measurement of results method
9Details of the outcome assessor
10Blinding
11Statistical analysis
12Results presentation


The assessment of bias risk determines the degree to which a study's design and methodology are free from any potential bias that could have an impact on the study's findings. Thus, determining the genuine impact of the test carried out from a specific application requires evaluating the risk of bias in a study [32].

Data Extraction and Synthesis

The eligibility of the publications was evaluated independently by three reviewers by looking at their titles, abstracts, and complete texts. Any differences or disagreements were discussed or settled with the assistance of the other two reviewers. A typical data extraction form (Table 2) with the following details was used to extract the research findings that were included:

Table 2 . Information extracted from the included studies..

Authors (year)Research typeType(s) of microorganismsEffective concentrationEvaluation methodResults
Zaid et al. (2015) [29]In vitroProtozoan: Plasmodium falciparumDrug sensitivity assay: IC50 =1039.3 ± 45.76 nM, IC90 = 2102.4 ± 56.2 nM Merozoite invasion assay: > 125 nMConventional malaria drug sensitivity assay and merozoite invasion assayAndrographolide's anti-plasmodi um action was less pronounced in the drug sensitivity assay, although it significantly (p < 0.05) decreased the proportion of RBCs with plasmodium ring infection in the merozoite invasion experiment.
Komaikul et al. (2023) [16]In vitroVirus: Human coronavirus (HCoV-OC43)Ranges between 0.92 mg/ml - 2.34 mg/mlCell viability and in-cell ELISA testsThe anti-HCoV-OC43 action of the 2.34 mg/ml andrographolidecontaining methanolic extract was less compared to DES extracts with less andrographolide contents of 0.92 to 1.46 mg/ml.
Yadav et al. (2022) [28]Ex vivoProtozoan: Bovine filarial parasite (Setaria cervi)IC50 value of 24.80 μMMTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] assayWith an IC50 value of 24.80 μM determined by the MTT experiment, andrographolide demonstrated potential antifilarial activity.
Paemanee et al. (2019) [18]In vitroVirus: Dengue virus (DENV)50 μMCells lines culture and standard plaque assayA significant reduction in the virus titer in response to andrographolide treatment was revealed with 50 μM concentration.
Arifullah et al. (2013) [24]In vitroBacteria/Fungi: Escherichia coli, Bacillus subtilis, Mycobacterium smegmatis, Staphylococcus aureus, Streptococcus thermophilus Pseudomonas aeruginosa, and Klebsiella pneumoniae Fungi (Yeasts): Candida albicans, Saccharomyces cerevisiae50 μg/ml- 350 μg/mlModified agar-disc diffusion method and broth microdilution methodAndrographolide demonstrated a broad range of growth inhibition activity with MIC values range between 50 μg/ml to 350 μg/ml against the bacteria Bacillus subtilis, Staphylococcus aureus, Streptococcus thermophilus, Escherichia coli, Mycobacterium smegmatis, Klebsiella pneumoniae, and Pseudomonas aeruginosa. When tested on the yeasts Candida albicans and Saccharomyces cerevisiae, it proved ineffective.
Panraksa et al. (2017) [19]In vitroVirus: Dengue virus (DENV)21.304 μM and 22.739 μMCells lines culture and standard plaque assay.Both the 21.304 μM and 22.739 μM 50% effective doses (EC50) of andrographolide demonstrated anti-DENV activity.
Bassey et al. (2021) [26]In vitroBacteria: Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Streptococcus pyogenes, and Staphylococcus aureus.60 μg/ml - 250 μg/mlBroth microdilution methodNeisseria gonorrhoeae was shown to be markedly susceptible to andrographolide (CF6) with 60 μg/ ml (MIC value), whereas S. pyogenes and S. aureus displayed 125 μg/ml MIC value each. At a concentration of 250 μg/ml, bacteria that are gram-negative, including E. coli, K. pneumonia, and P. aeruginosa were all suppressed.
Wang et al. (2018) [15]In vitroVirus: Enterovirus D68 (EV-D68)EC50 = 3.45 μMCell culture, cell viability assay, and endpoint dilution assay (EPDA).With an EC50 of 3.45 μM, andrographolide greatly reduced EV-D68 RNA replication and had antiviral efficacy against EV-D68 infection.
Nidiry et al. (2015) [27]In vitroFungi: Alternaria solani500 mg/lSpore germination inhibition study.At a 500 mg/l concentration, andrographolide inhibits Alternaria solani spore germination by 64.8%.
Theerawata nasirikul et al. (2022) [21]In vitroVirus: Foot-and-mouth-disease virus (FMDV)52.18 ± 0.01 μMCell culture, antiviral activity assays, and RTqPCR.Effective concentration (EC50) value of 52.18 ± 0.01 μM was observed for andrographolide's inhibitory activity as measured by RT-qPCR.
Lee et al. (2014) [17]In vitroVirus: Hepatitis C virus (HCV)6.0 ± 0.5 μMCell culture, western blotting, and qRT-PCR analysis.With an effective concentration (EC50) value of 6.0 ± 0.5 μM, andrographolide considerably decreased HCV RNA levels.
Ali and Ahmad Mir (2020) [23]In vitroBacteria: E. coli MTCC16793.0 mg/mlAgar well diffusion method for antibacterial testing.The bacterial pathogen (Escherichia coli MTCC1679) was inhibited by the andrographolide compound with a 12 ± 1.0 mm zone of inhibition at a concentration of 3.0 mg/ml.
Gopalakris hnan et al. (2016) [33]In vitroProtozoan: Theileria equiHighest concentrati on was 100 μMIn vitro growth inhibition assay.Andrographolide was ineffective in inhibiting the growth of T. equi parasites even at the highest concentration of 100 μM used.
Shi et al. (2020) [20]In vitroViruses: 2019 novel corona virus (2019-nCoV) and severe acute respiratory syndrome coronavirus (SARS-CoV)2019-nCoV: 15.05 ± 1.58 μM SARS-CoV: 5.00 ± 0.67 μMProtease activity assay.The 2019-nCoV Mpro protease activity was inhibited by andrographolide at an IC50 of 15.05 ± 1.58 μM, while SARS-CoV Mpro was suppressed at an IC50 of 5.00 ± 0.67 μM.
Ativui et al. (2022) [25]In vitroBacteria: Salmonella Paratyphi B, Enterococcus faecalis, S. pyogenes, E. coli, Vibrio cholerae, Pseudomonas aeruginosa, Staphylococcus aureus, Proteus mirabilis, Klebsiella pneumoniae Fungus (Yeast): C. albicans1.71 μg/ml and 875 μg/mlHighthroughput spot culture growth inhibition (HTSPOTi) technique (anti-infective assay).While resistance was observed in Staphylococcus aureus, Proteus mirabilis, Enterococcus faecalis, and Candida albicans even at 875 μg/ml concentration, andrographolide exhibited bactericidal activity against Klebsiella pneumoniae, Streptococcus pyogenes, Salmonella paratyphi B, with the maximum inhibitory activity of 1.71 μg/ml against Vibrio cholerae.
Wintachai et al. (2015) [22]In vitroVirus: chikungunya virus (CHIKV)EC50 value of 77 μMCell lines culture, virucidal and standard plaque assays.With an effective concentration (EC50) of 77 μM, andrographolide showed good suppression of CHIKV infection and decreased virus production by about 3 log10.


i. Author(s) and publication year.

ii. Type of research (in vitro or ex vivo).

iii. Type of microorganism (bacteria, fungi, viruses, or protozoa).

iv. Effective concentration.

v. Method of evaluation and results (i.e., the antimicrobial activity or effect).

Overview of the Included Studies

This comprehensive review included 16 studies overall (15 studies conducted in vitro and one ex vivo study). The studies evaluated andrographolide's antimicrobial action against a range of microorganisms, such as viruses, bacteria, fungi, and protozoa (Table 2). Of these studies, two assessed andrographolide's antimicrobial efficacy against bacteria [23, 26], two examined its antimicrobial activity against both bacteria and fungi [24, 25], one study evaluated its antimicrobial activity against only a fungus [27], eight studies investigated its antimicrobial activity against viruses [15-22], and three studies evaluated its antimicrobial activity against protozoa [28, 29, 33].

Efficacy of Andrographolide against Different Microorganisms

This review evaluated the antimicrobial activity of andrographolide against a broad variety of microbes, such as viruses, protozoa, bacteria, and fungi. Most of the studies (Table 2) reported that andrographolide exhibited significant antimicrobial activity against various strains of bacterial species, including Staphylococcus aureus, Streptococcus thermophilus, Bacillus subtilis, Mycobacterium smegmatis, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Neisseria gonorrhoeae, Streptococcus pyogenes, Salmonella paraty-phi B, and Vibrio cholerae [23-26].

However, Proteus mirabilis, Enterococcus faecalis, and Staphylococcus aureus were found to be resistant to it even at high concentration as established in the study conducted by [25]. Andrographolide was also found to be effective against the fungus Alternaria solani [27], but was found ineffective against Saccharomyces cerevisiae and Candida albicans [24, 25]. In addition, andrographolide showed antiviral activity against several viruses, including Human Coronavirus, Dengue virus, Enterovirus D68, Foot-and-Mouth-Disease virus, Hepatitis C virus, Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV), 2019 Novel Coronavirus (2019-nCoV), and Chikungunya virus [15-22]. Furthermore, andrographolide exhibited antiprotozoal activity against certain protozoa, including Plasmodium falciparum and Setaria cervi [28, 29], but was found to be ineffective against Theileria equi [33]. Depending on the microbial strain and the experimental settings, andrographolide's antimicrobial effectiveness varies against several types of microbes.

Quality of the Included Studies

The quality of each study was assessed using the Quality Evaluation Instrument for In Vitro Studies [31]. This tool was utilized to evaluate the bias risk of individual studies included in the current review. Out of the sixteen (16) studies that make up this review (Table 3), 11 (68.75%) were evaluated to have less bias risk with the percentage score range of 78.57%-92.86% [15-22, 24, 28, 29], whereas 5 (31.25%) studies had medium risk of bias with the percentage score of between 57.14% and 64.29% [23, 25-27, 33]. There was no significant bias risk associated with any of the examined studies. This suggests that the overall caliber of the research covered in this review was acceptable.

Table 3 . Quality assessment and risk of bias detection for the included studies..

AuthorsCriteria/check lists
123456789101112Overall scoreScore (%)Risk of bias
Zaid et al. (2015) [29]2--12--2-1121178.57Low
Komaikul et al. (2023) [16]2--12--2-1221285.71Low
Yadav et al. (2022) [28]2--12--2-1221285.71Low
Paemanee et al. (2019) [18]2--22--2-1221392.86Low
Arifullah et al. (2013) [24]2--22--2-1221392.86Low
Panraksa et al. (2017) [19]2--22--2-0221285.71Low
Bassey et al. (2021) [26]0--12--2-112964.29Medium
Wang et al. (2018) [15]1--12--2-1221178.57Low
Nidiry et al. (2015) [27]1--12--2-012964.29Medium
Theerawatanasirikul et al. (2022) [21]1--22--2-1121178.57Low
Lee et al. (2014) [17]1--22--2-1221285.71Low
Ali and Ahmad Mir (2020) [23]0--22--2-002857.14Medium
Gopalakrishnan et al. (2016) [33]1--22--2-002964.29Medium
Shi et al. (2020) [20]2--02--2-1221178.57Low
Ativui et al. (2022) [25]2--02--2-002857.14Medium
Wintachai et al. (2015) [22]0--22--2-1221178.57Low

Unapplicable items not included in the computation (-)..


Summary and Overall Outcome of the Review

Andrographolide has been shown in numerous studies to have antibacterial action against a variety of harmful bacteria. One study [24] revealed that at minimum inhibitory concentration (MIC) ranges of 100-350 μg/ml, andrographolide demonstrated a broad range of growth inhibition activity against Bacillus subtilis, Escherichia coli, Mycobacterium smegmatis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus thermophilus. Another study [26] uncovered the antibacterial activity of andrographolide against Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus pyogenes at MIC values ranging between 125 μg/ml and 250 μg/ml. These studies also showed that Neisseria gonorrhoeae was significantly susceptible to andrographolide at an MIC value of 60 μg/ml. In addition, a study by [23] confirmed that at a dosage of 3.0 mg/ml, andrographolide displayed antibacterial activity against Escherichia coli. Conversely, the study of [25] established that Proteus mirabilis, Enterococcus faecalis, and Staphylococcus aureus were resistant to andrographolide, even at the highest concentration of 875 μg/ml. However, they also noted in the same investigation that andrographolide demonstrated bactericidal and inhibitory effects on Salmonella enterica ser. Paratyphi B, Streptococcus pyogenes, Klebsiella pneumoniae, and Vibrio cholerae.

According to the review's findings, there is a dearth of research on andrographolide's antifungal activity since the compound's effectiveness against fungi has only been looked at in a limited number of studies. However, [24] reported in their investigation that andrographolide had no effect on the yeasts Candida albicans and Saccharomyces cerevisiae. This was supported by [25], who also established that C. albicans was resistant to andrographolide, even at the concentration of 875 μg/ml. In a contrary report, andrographolide was found to exhibit inhibitory activity against Alternaria solani spore germination by 64.8% at a concentration of 500 mg/l [27]. This implies that more research is needed on the antifungal activity of andrographolide.

Andrographolide was found to exhibit antiviral activity against several viruses. It suppressed the protease activities of 2019-nCoV Mpro and SARS-CoV Mpro at the half-maximal inhibitory concentration (IC50) values of 15.05 ± 1.58 μM, and 5.00 ± 0.67 μM, respectively [20]. This was corroborated by the findings of [16], who also reported the anti-Human coronavirus (anti-HCoV-OC43) activity of andrographolide at concentrations between 0.92 mg/ml and 2.34 mg/ml. Moreover, a study by [19] recorded that andrographolide demonstrated anti-dengue virus (anti-DENV) activity at the half-maximal effective concentration (EC50) values of 21.304 μM and 22.739 μM. This was also recorded by [18], who found a significant reduction in dengue virus (DENV) titer in response to andrographolide treatment at 50 μM concentration. In addition, andrographolide was found to greatly reduce Enterovirus D68 (EV-D68) RNA replication and had antiviral efficacy against EV-D68 infection at the EC50 value of 3.45 μM [15]. The research of [21] reported that andrographolide demonstrated inhibitory action against the virus causing foot-and-mouth disease (FMDV) at the median effective dose value (ED50) of 52.18 ± 0.01 μM. Also reported by [17], andrographolide considerably decreased hepatitis C virus (HCV) RNA levels at an EC50 value of 6.0 ± 0.5 μM. Furthermore, a study by [22] established that the compound demonstrated good suppression of infection with chikungunya virus (CHIKV) and decreased the virus production at an EC50 value of 77 μM.

In this review, andrographolide has also been associated with antiprotozoal activity against a few protozoan parasites. As reported by [29], andrographolide was found to significantly (p < 0.05) decrease the proportion of red blood cells (RBCs) with Plasmodium ring infection despite its less pronounced anti-plasmodium action. Also, in corroboration of the antiprotozoal activity of andrographolide, [28] established that the compound demonstrated potential anti-filarial activity against Setaria cervi (a bovine filarial parasite) at an IC50 value of 24.80 μM. On the other hand, andrographolide was found to be ineffective against Theileria equi parasites, even at the highest concentration of 100 μM [33].

According to the findings of this thorough review, andrographolide may be useful as a natural antimicrobial agent against various microorganisms, despite some differences recorded in the results of the included studies. These differences could be due to a variety of factors, such as strain variances, compound potency, methodological approach, and other variables.

Conclusion

Andrographolide appears to have strong antimicrobial activity against a variety of microbial strains, as we have reported in this review according to the available information. This compound is a viable candidate for the development of new therapeutic drugs due to its ability to reduce and hinder the growth and activity of bacterial pathogens, some fungal and protozoal pathogens, and the replication and proliferation of viral pathogens. However, additional study is required to assess and clarify andrographolide’s efficacy against a wide variety of fungal pathogens, such as Aspergillus species, Trichophyton species, Microsporum species, among others, as only a few studies were found on the antifungal activity of the compound, and these were limited to only C. albicans, S. cerevisiae, and Alternaria solani. Additionally, more studies are needed to investigate the mechanisms of action, toxicity, and safety of andrographolide in clinical trials, as well as its potential side effects and drug interactions.

Acknowledgments

We thank the Malaysia Ministry of Higher Education for funding this research work through the Fundamental Research Grant Scheme under Grant No. FRGS/1/2019/SKK11/USM/02/3.

Conflict of Interest

The authors have no financial conflicts of interest to declare.

Fig 1.

Figure 1.PRISMA flow diagram of articles screened for inclusion in the systematic review.
Journal of Microbiology and Biotechnology 2025; 35: https://doi.org/10.4014/jmb.2408.08028

Table 1 . The score distribution and evaluation parameters for bias risk in the included studies..

S/NCriteria/Check list
Adequately stated (Score = 2)Inadequately stated (Score = 1)Unspecified (Score = 0)Unapplicable (Excluded)
1Clearly defined aims or objectives
2A thorough description of the sample size calculation
3Detailed description of sampling method
4Information about the comparison group
5Detailed description of the methodology
6Operator details
7Randomization
8Measurement of results method
9Details of the outcome assessor
10Blinding
11Statistical analysis
12Results presentation

Table 2 . Information extracted from the included studies..

Authors (year)Research typeType(s) of microorganismsEffective concentrationEvaluation methodResults
Zaid et al. (2015) [29]In vitroProtozoan: Plasmodium falciparumDrug sensitivity assay: IC50 =1039.3 ± 45.76 nM, IC90 = 2102.4 ± 56.2 nM Merozoite invasion assay: > 125 nMConventional malaria drug sensitivity assay and merozoite invasion assayAndrographolide's anti-plasmodi um action was less pronounced in the drug sensitivity assay, although it significantly (p < 0.05) decreased the proportion of RBCs with plasmodium ring infection in the merozoite invasion experiment.
Komaikul et al. (2023) [16]In vitroVirus: Human coronavirus (HCoV-OC43)Ranges between 0.92 mg/ml - 2.34 mg/mlCell viability and in-cell ELISA testsThe anti-HCoV-OC43 action of the 2.34 mg/ml andrographolidecontaining methanolic extract was less compared to DES extracts with less andrographolide contents of 0.92 to 1.46 mg/ml.
Yadav et al. (2022) [28]Ex vivoProtozoan: Bovine filarial parasite (Setaria cervi)IC50 value of 24.80 μMMTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] assayWith an IC50 value of 24.80 μM determined by the MTT experiment, andrographolide demonstrated potential antifilarial activity.
Paemanee et al. (2019) [18]In vitroVirus: Dengue virus (DENV)50 μMCells lines culture and standard plaque assayA significant reduction in the virus titer in response to andrographolide treatment was revealed with 50 μM concentration.
Arifullah et al. (2013) [24]In vitroBacteria/Fungi: Escherichia coli, Bacillus subtilis, Mycobacterium smegmatis, Staphylococcus aureus, Streptococcus thermophilus Pseudomonas aeruginosa, and Klebsiella pneumoniae Fungi (Yeasts): Candida albicans, Saccharomyces cerevisiae50 μg/ml- 350 μg/mlModified agar-disc diffusion method and broth microdilution methodAndrographolide demonstrated a broad range of growth inhibition activity with MIC values range between 50 μg/ml to 350 μg/ml against the bacteria Bacillus subtilis, Staphylococcus aureus, Streptococcus thermophilus, Escherichia coli, Mycobacterium smegmatis, Klebsiella pneumoniae, and Pseudomonas aeruginosa. When tested on the yeasts Candida albicans and Saccharomyces cerevisiae, it proved ineffective.
Panraksa et al. (2017) [19]In vitroVirus: Dengue virus (DENV)21.304 μM and 22.739 μMCells lines culture and standard plaque assay.Both the 21.304 μM and 22.739 μM 50% effective doses (EC50) of andrographolide demonstrated anti-DENV activity.
Bassey et al. (2021) [26]In vitroBacteria: Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Streptococcus pyogenes, and Staphylococcus aureus.60 μg/ml - 250 μg/mlBroth microdilution methodNeisseria gonorrhoeae was shown to be markedly susceptible to andrographolide (CF6) with 60 μg/ ml (MIC value), whereas S. pyogenes and S. aureus displayed 125 μg/ml MIC value each. At a concentration of 250 μg/ml, bacteria that are gram-negative, including E. coli, K. pneumonia, and P. aeruginosa were all suppressed.
Wang et al. (2018) [15]In vitroVirus: Enterovirus D68 (EV-D68)EC50 = 3.45 μMCell culture, cell viability assay, and endpoint dilution assay (EPDA).With an EC50 of 3.45 μM, andrographolide greatly reduced EV-D68 RNA replication and had antiviral efficacy against EV-D68 infection.
Nidiry et al. (2015) [27]In vitroFungi: Alternaria solani500 mg/lSpore germination inhibition study.At a 500 mg/l concentration, andrographolide inhibits Alternaria solani spore germination by 64.8%.
Theerawata nasirikul et al. (2022) [21]In vitroVirus: Foot-and-mouth-disease virus (FMDV)52.18 ± 0.01 μMCell culture, antiviral activity assays, and RTqPCR.Effective concentration (EC50) value of 52.18 ± 0.01 μM was observed for andrographolide's inhibitory activity as measured by RT-qPCR.
Lee et al. (2014) [17]In vitroVirus: Hepatitis C virus (HCV)6.0 ± 0.5 μMCell culture, western blotting, and qRT-PCR analysis.With an effective concentration (EC50) value of 6.0 ± 0.5 μM, andrographolide considerably decreased HCV RNA levels.
Ali and Ahmad Mir (2020) [23]In vitroBacteria: E. coli MTCC16793.0 mg/mlAgar well diffusion method for antibacterial testing.The bacterial pathogen (Escherichia coli MTCC1679) was inhibited by the andrographolide compound with a 12 ± 1.0 mm zone of inhibition at a concentration of 3.0 mg/ml.
Gopalakris hnan et al. (2016) [33]In vitroProtozoan: Theileria equiHighest concentrati on was 100 μMIn vitro growth inhibition assay.Andrographolide was ineffective in inhibiting the growth of T. equi parasites even at the highest concentration of 100 μM used.
Shi et al. (2020) [20]In vitroViruses: 2019 novel corona virus (2019-nCoV) and severe acute respiratory syndrome coronavirus (SARS-CoV)2019-nCoV: 15.05 ± 1.58 μM SARS-CoV: 5.00 ± 0.67 μMProtease activity assay.The 2019-nCoV Mpro protease activity was inhibited by andrographolide at an IC50 of 15.05 ± 1.58 μM, while SARS-CoV Mpro was suppressed at an IC50 of 5.00 ± 0.67 μM.
Ativui et al. (2022) [25]In vitroBacteria: Salmonella Paratyphi B, Enterococcus faecalis, S. pyogenes, E. coli, Vibrio cholerae, Pseudomonas aeruginosa, Staphylococcus aureus, Proteus mirabilis, Klebsiella pneumoniae Fungus (Yeast): C. albicans1.71 μg/ml and 875 μg/mlHighthroughput spot culture growth inhibition (HTSPOTi) technique (anti-infective assay).While resistance was observed in Staphylococcus aureus, Proteus mirabilis, Enterococcus faecalis, and Candida albicans even at 875 μg/ml concentration, andrographolide exhibited bactericidal activity against Klebsiella pneumoniae, Streptococcus pyogenes, Salmonella paratyphi B, with the maximum inhibitory activity of 1.71 μg/ml against Vibrio cholerae.
Wintachai et al. (2015) [22]In vitroVirus: chikungunya virus (CHIKV)EC50 value of 77 μMCell lines culture, virucidal and standard plaque assays.With an effective concentration (EC50) of 77 μM, andrographolide showed good suppression of CHIKV infection and decreased virus production by about 3 log10.

Table 3 . Quality assessment and risk of bias detection for the included studies..

AuthorsCriteria/check lists
123456789101112Overall scoreScore (%)Risk of bias
Zaid et al. (2015) [29]2--12--2-1121178.57Low
Komaikul et al. (2023) [16]2--12--2-1221285.71Low
Yadav et al. (2022) [28]2--12--2-1221285.71Low
Paemanee et al. (2019) [18]2--22--2-1221392.86Low
Arifullah et al. (2013) [24]2--22--2-1221392.86Low
Panraksa et al. (2017) [19]2--22--2-0221285.71Low
Bassey et al. (2021) [26]0--12--2-112964.29Medium
Wang et al. (2018) [15]1--12--2-1221178.57Low
Nidiry et al. (2015) [27]1--12--2-012964.29Medium
Theerawatanasirikul et al. (2022) [21]1--22--2-1121178.57Low
Lee et al. (2014) [17]1--22--2-1221285.71Low
Ali and Ahmad Mir (2020) [23]0--22--2-002857.14Medium
Gopalakrishnan et al. (2016) [33]1--22--2-002964.29Medium
Shi et al. (2020) [20]2--02--2-1221178.57Low
Ativui et al. (2022) [25]2--02--2-002857.14Medium
Wintachai et al. (2015) [22]0--22--2-1221178.57Low

Unapplicable items not included in the computation (-)..


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