Abstract

Background

   Rhizomes of Curcuma wanenlueanga Saensouk, Thomudtha & Boonma have been
   used in Thai traditional medicine and are included as an ingredient in
   a Thai traditional liniment formula listed by the Ministry of Public
   Health of Thailand for treating symptoms related to joints and muscle
   inflammation. However, anti-inflammatory activity and bioactive
   constituents of C. wanenlueanga have not yet been investigated. Thus,
   this study aimed to investigate the anti-inflammatory activity and
   underlying mechanism of C. wanenlueanga rhizome extract and its
   responsible bioactive components.

Methods

   The fractionation of bioactive compounds from C. wanenlueanga extract
   was guided by antioxidant activity on DPPH and Griess assays, and
   anti-inflammatory activity on LPS-induced RAW 264.7 cells. The
   biological activities of isolated compounds were first predicted by
   network pharmacology and further confirmed in cell-based assay with
   LPS-induced RAW 264.7 cells and enzyme-linked immunosorbent assay
   (ELISA) of tumor necrosis factor alpha (TNF-α) and interleukin-6
   (IL-6).

Results

   The ethanolic extract of C. wanenlueanga rhizomes was proved to show
   anti-inflammatory activity on LPS-induced RAW 264.7 cells. Three
   curcuminoids including curcumin (1), demethoxycurcumin (2),
   dihydrodemethoxycurcumin (3), and two sesquiterpenoids, namely
   curcumenone (4), and zedoarondiol (5) were separated from
   anti-inflammatory fractions of C. wanenlueanga extract. The
   anti-inflammatory activity of these compounds to attenuate the
   productions of TNF-α and IL-6 was predicted by pharmacological network.
   The inflammatory assays, including the Griess assay for NO and ELISA
   for TNF-α and IL-6, confirmed that all isolated compounds reduced the
   production of these inflammatory mediators.

Conclusion

   The present study shows the accordance between the results from
   pharmacological network and cell-based assays, which indicate the
   anti-inflammatory activity of C. wanenlueanga rhizomes and their
   bioactive constituents. This suggests the potential of bioactive
   compounds 1–5 to be used for quality assessment of C. wanenlueanga
   extract in the development of herbal products.

Supplementary Information

   The online version contains supplementary material available at
   10.1186/s12906-025-04884-0.

   Keywords: Curcuma wanenlueanga, Zingiberaceae, Anti-inflammatory
   activity, Bioassay-guided isolation, Network pharmacology, Bioactive
   marker

Background

   Inflammation is a defense mechanism of the human immune system in
   response to irritants, typically leading to heat, swelling and redness
   of the affected area [[36]1]. Acute inflammation occurs after injury
   and normally lasts for a few days to a week. If it does not resolve in
   time, it will progress to chronic inflammation in around six weeks,
   leading to the tissue damage and change in tissue integrity [[37]2]. In
   an inflammation process, inflammatory cells such as neutrophils and
   macrophages migrate to the site of inflammation [[38]3], with a
   secretion of key cytokines including interleukin-1 (IL-1),
   interleukin-2 (IL-2), interleukin-6 (IL-6), tumor necrosis factor alpha
   (TNF-α) and interferon gamma (IFN-γ) [[39]4]. Inflammation can occur in
   every part of the body, and some related conditions, such as diabetes,
   cardiovascular disease and allergies, can be life-threatening.
   Additionally, muscle and joint inflammatory diseases, including
   myositis, gout, and rheumatoid arthritis (RA), cause discomfort, severe
   pain, permanent loss of functions, and critically affect all aspects of
   patient’s quality of life. Several approaches are applied to treat
   muscle and joint inflammatory diseases, such as physical therapy,
   medication and surgery [[40]5]. The first method is safe but takes a
   long time and is sometimes not medically effective. Surgery is an
   extreme method to deal with terminal conditions. Though, this method
   can lead to irreversible adverse effects. Nonsteroidal
   anti-inflammatory drugs (NSAIDs) are effective drugs that reduce pain,
   inflammation, and fever. Nevertheless, they have some potential adverse
   effects, including ulcers and bleeding in the gastrointestinal tract
   [[41]6], elevated blood pressure [[42]7], risk of heart attack [[43]8],
   kidney and liver damages [[44]9].

   Recently, herbal medicine has gained attention as an alternative way to
   alleviate muscle and joint inflammation. Several medicinal plants in
   the curcuma genus, such as turmeric (Curcuma longa L.) [[45]10],
   Curcuma aromatica Salisb [[46]11]. and Curcuma latifolia Roscoe
   [[47]12], have been proved to relieve inflammation. As the growing use
   of herbal medicine for treatment of various diseases [[48]13], studies
   on medicinal plants to deal with inflammatory diseases remain an
   interesting area of research worldwide. Providentially, network
   pharmacology has been established to explain the relationship among
   chemical constituents in plants, drug targets, signaling pathways and
   diseases in holistic way [[49]14]. For instance, Shi and colleagues
   explored the anti-inflammatory and antioxidant constituents of Solanum
   melongena L. and investigated their mechanisms using network
   pharmacology [[50]15]. Limcharoen and coworkers utilized network
   pharmacology to disclose the ability of (−)-dendroparishiol from
   Dendrobium parishii to manage bacterial meningitis-associated
   neuroinflammation [[51]16]. These support the potential of network
   pharmacology, which helps scientists open a new possibility to discover
   useful medicinal herbs and their bioactive constituents.

   Curcuma wanenlueanga Saensouk, Thomudtha & Boonma (Fig. [52]1A) is a
   plant species in the Zingiberaceae family. It distributes and is
   cultivated through northern part of Thailand. This perennial herb could
   be up to 150 centimeters tall and has ovoid rhizome with internal
   yellow with darker core. The specific epithet of this plant was derived
   from its Thai vernacular name “Wan-en-lueang”, which mean “yellow herb
   related to tendons” [[53]17]. In Thailand, C. wanenlueanga rhizomes
   have been recognized in several documents for their traditional use
   (Fig. [54]1B), including treatment of beriberi related to tendon,
   paralysis, sprains, bruise, swelling, backache, joint pain and
   tendonitis [[55]18]. C. wanenlueanga is also listed as an ingredient in
   a Thai traditional liniment formula certified by the Ministry of Public
   Health of Thailand [[56]19]. There is a previous study revealed
   antioxidant, anti α-glucosidase activities C. wanenlueanga [[57]20].
   However, preliminary investigation of anti-inflammatory activity and
   chemical constituents that are responsible for the bioactivity of C.
   wanenlueanga has not been investigated. Thus, this study aimed to
   investigate the anti-inflammatory activity of C. wanenlueanga rhizomes.
   Bioassay-guided fractionation was used to obtain bioactive
   constituents. The targets and mechanisms of action of isolated
   compounds were then forecasted by network pharmacology and confirmed by
   enzyme-linked immunosorbent assay (ELISA).

Fig. 1.

   [58]Fig. 1
   [59]Open in a new tab

   Curcuma wanenlueanga Saensouk, Thomudtha & Boonma and its
   ethnobotanical references. (A) Morphological features of a fresh