Xu et al

Tropical Journal of Pharmaceutical Research April 2024; 23 (4): 691-696

ISSN: 1596-5996 (print); 1596-9827 (electronic)

Available online at http://www.tjpr.org

http://dx.doi.org/10.4314/tjpr.v23i4.3
Original Research Article

Biological activity and mechanism of action of Gotu kola

oil in skin wound repair
Biao Xu*, Yushun Duan, Deshun Yu, Panpan Jia
Department of Emergency (Trauma), Shulan (Hangzhou) Hospital Affiliated to Zhejiang Shuren University Shulan International
Medical College, Hangzhou, Zhejiang Province 310022, China

*For correspondence: Email: xianb979@163.com

Sent for review: 7 December 2023 Revised accepted: 25 March 2024

Abstract
Purpose: To investigate the biological activity and mechanism of action of Gotu Kola oil in skin wound
repair.
Methods: In this study, a total of 40 Sprague-Dawley (SD) rats were meticulously chosen to establish a
skin wound model. The rats were randomly allocated to a study group, which received topical
application of Gotu kola oil, and control group, which served as the model control. Each group consisted
of 20 rats to ensure an adequate sample size. The wound area was measured using precise digital
imaging software to quantitatively determine the rate of wound healing. Expression levels of hepatocyte
growth factor (HGF), vascular endothelial growth factor (VEGF) and transforming growth factor-beta 1
(TGF-β1) were determined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR)
while the extent of microvassel regeneration at the wound site was determined by immunohistochemical
method using CD34 as a marker.
Results: The wound healing rate in the study group was higher than in the control group. In the study
group, the transcription levels of HGF and VEGF were higher than in the control group on days 3, 7, and
14, while the transcription level of TGF-β1 was higher on days 7 and 14. Microvessel density (MVD) in
the study group was higher than in the control group on days 3 and 7 (p < 0.05).
Conclusion: Gotu kola oil facilitates wound healing by promoting the generation of capillaries at the
wound site. Harnessing this potential of Gotu kola oil may lead to the development of more efficacious
treatment strategies for skin wound repair, ultimately enhancing the quality of life of the patients.

Keywords: Gotu kola oil, Skin wound, Repair, Biological activity, Mechanism of action

This is an Open Access article that uses a funding model which does not charge readers or their institutions
for access and distributed under the terms of the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative
(http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly credited.

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INTRODUCTION Skin injuries, resulting from various causes, are

common and significant health concerns that
The skin is one of the largest organs in the profoundly affect patients' physical well-being
human body and its integrity is crucial for and quality of life. Skin trauma refers to damage
maintaining physiological functions and or harm inflicted upon the skin or underlying
defending against external environmental tissues due to physical trauma, irritation, or
threats. external factors. It encompasses a wide range of
injuries, such as cuts, tears, abrasions, burns,

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© 2024 The authors. This work is licensed under the Creative Commons Trop J Pharm4.0
Attribution Res, April 2024;License
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bruises, punctures, ulcers, and others. Different Animal modeling

types of skin trauma can lead to varying degrees
of damage, spanning from minor abrasions to The experiment began after a one-week
severe burns or deep wounds [1,2]. Following acclimatization period in a standard animal
trauma, the skin initiates a self-repair mechanism laboratory environment. To induce circular skin
to expedite its restoration. Consequently, defects with a 10 mm diameter, a dermatological
exploring effective approaches to promote skin biopsy punch was employed. This standardized
wound repair and healing has been a paramount instrument allows for the precise and controlled
focus of clinical medicine and biomedical study. removal of tissue, ensuring consistent
dimensions in the circular wounds. The defects
In recent decades, extensive investigations have were created on both the left and right sides of
elucidated the molecular and cellular the skin on the back of the rats' second lumbar
mechanisms underlying skin wound repair, vertebra. Subsequently, all rats were randomly
aiming to identify potential therapeutic strategies. assigned to two groups using a random number
However, despite these efforts, an ideal table method.
treatment modality that simultaneously enhances
healing speed, improves healing quality and Treatment method
reduces complications remains elusive.
Consequently, the exploration of novel treatment Both groups underwent wound treatment twice
approaches and efficacious medications has daily at scheduled intervals. Study group
become a pivotal area of current study. received Gotu Kola oil administered as follows:
480 milligrams dissolved in 200 microliters of
Gotu Kola oil, derived from natural sources, has DMSO, thoroughly mixed to achieve a uniform
garnered significant study interest in recent and stable solution. Subsequently, 20 microliters
years. It has been widely utilized in traditional of the solution were extracted and combined with
medical systems, especially in Asian regions, for 16 grams of white Vaseline. The mixture was
the management of various skin conditions heated until melted, then cooled to produce a
including burns, ulcers, trauma and inflammatory blue, semi-transparent, viscous paste. The
skin diseases [3,4]. Concurrently, modern resulting ointment, containing 3 milligrams of
scientific study has gradually unraveled the blue oil per gram, was meticulously applied to
potential roles of Gotu Kola oil in skin wound cover the wound, ensuring complete coverage of
repair. Therefore, the objective of this study is to the wound site. In contrast, control group was
elucidate the biological activity and mechanisms treated with Vaseline ointment containing DMSO
of action of Gotu Kola oil in the context of skin applied to the wound.
wound repair. Through comprehensive
experimentation and molecular biology Evaluation of parameters/indices
methodologies, this study comprehensively
investigates the effects of Gotu Kola oil on
wound sites, encompassing its potential impact Wound healing parameters
on healing quality, vascular regeneration and
other relevant factors. This in-depth exploration Photographs and samples were obtained at
of these critical facets aims to provide scientific various time points, including the day of
evidence for the development of more effective successful modeling and at days 3, 7, 14 and 21.
strategies for skin wound repair treatments, ImageJ software was employed to measure the
ultimately enhancing patients' quality of life and wound area (WA). The healing rate (HR) was
expediting their recovery process. calculated using Eq 1.

EXPERIMENTAL HR = (WAp/WAi)100 …………….. (1)

Animals where WAp: wound area at the time of

photography; WAi : wound area (initial). This
Forty male Sprague-Dawley (SD) rats, weighing method allowed for an accurate assessment of
between 240 and 260 g were chosen for this the progression of wound healing over the
study. These rats demonstrated normal body specified time intervals.
temperature, respiration and pulse rates. After
the successful modeling process, they were Gene expression analysis
randomly allocated into two groups: study group,
consisting of 20 rats receiving Gotu Kola oil Transcription levels of HGF, VEGF and TGF-β1
treatment, and control group, comprising 20 rats were determined using quantitative Reverse
serving as the model control group. Transcription-Polymerase Chain Reaction (qRT-

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 Xu et al

PCR). Biological samples, comprising RNA considered statistically significant when the p-
extracted from tissue specimens, underwent a value was less than 0.05.
systematic processing method for gene
expression analysis. Following RNA extraction RESULTS
and purification, the RNA was reverse
transcribed into complementary DNA (cDNA). Healing rate
The resulting cDNA served as a template for
quantitative reverse transcription-polymerase At the 7th and 14th days, the wound healing rate
chain reaction (qRT-PCR). of study group rats was higher than that of
control group (p < 0.05) as presented in Figure 1.
Immunohistochemical assessment of CD34-
negative marked blood vessels
Control group
Tissue samples were dewaxed, dehydrated, Study group
incubated using specific antibodies, stained and
clarified. Dewaxing involved the removal of 150

Wound healing rate （%）

paraffin from the tissue sections using a xylene,
while dehydration ensured the elimination of
residual water content. Paraffin sections were 100 *
prepared accordingly. At least, three random
200x magnification fields were selected from *
each group of sections for photography. The 50
average number of blood vessels in these fields
was calculated as the Microvessel Density
(MVD) value, representing the number of blood
vessels per high-power field (typically expressed 0
as number/field). This method facilitated the Day 3 Day 7 Day 14 Day 21
quantitative analysis of blood vessel density
within the tissue samples. Figure 1: Comparison of wound healing rates
between the two groups. *P < 0.05 vs. control group
Statistical analysis
Transcription levels of relevant growth
Graphs in this study were processed using factors
GraphPad Prism 8, while data organization and
analysis were conducted with SPSS 21.0 In study group rats, the transcription levels of
software. Quantitative data was presented as HGF and VEGF were higher than those in control
mean ± standard deviation and intergroup group on the 3rd day, transcription level of TGF-
comparisons were performed using the t-test. β1 was higher than that in control group on the
Count data was expressed as n (%) and 7th day (p < 0.05) as shown in Figure 2.
intergroup comparisons were analyzed using the
χ² test. Differences between groups were

Control group Control group Control group

Study group Study group Study group
6 5 10
* *
*
4 8
TGF-β1 mRNA
VEGF mRNA
HGF mRNA

4
3 6

2 4
2
1 2

0 0 0
Day 0 Day 3 Day 7 Day 14 Day 0 Day 3 Day 7 Day 14 Day 0 Day 3 Day 7 Day 14

Figure 2: Comparison of transcription levels of HGF, VEGF, and TGF-β1 between the two groups. *P < 0.05 vs.
control group

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 Xu et al

Wound microvascular density promote the healing of skin wounds. It stimulates

the synthesis of collagen, and accelerates the
In study group rats, the microvascular density repair and regeneration of damaged tissues.
(MVD) was higher than in control group on the Additionally, Gotu kola oil aids in increasing cell
3rd and 7th day and lower on the 14th day, while proliferation, thus facilitating the speedy recovery
the differences were insignificant (p > 0.05) as of wounds [8]. Some studies suggest that Gotu
depicted in Figure 3. Kola oil has antibacterial and antifungal activity,
which helps prevent or treat wound infections.
Control group
The antioxidant properties of Gotu Kola oil and
Study group its promotion of collagen make it a potentially
anti-aging compound. It helps maintain skin
150
elasticity and a youthful appearance. Gotu Kola
oil thus exhibits multiple biological activities,
including anti-inflammatory, antioxidant, wound-
100 healing promotion, antibacterial and anti-aging
effects. These characteristics have garnered
MVD

significant attention in the fields of skin health

and wound healing, and Gotu kola oil is widely
50 used in skincare products and medical
treatments [5,9].

The results indicate that Gotu kola oil possesses

0
significant biological activity and contributes to
Day 3 Day 7 Day 14
the promotion of skin wound healing. It was
observed that in groups treated with Gotu kola
Figure 3: Comparison of wound microvascular density oil, wound healing rate was significantly higher
between the two groups than in control group. This result is consistent
with previous study, demonstrating the positive
DISCUSSION role of Gotu kola oil in wound healing. This may
be attributed to the polysaccharide components
Gotu Kola oil (Centella asiatica), also known as of Gotu kola oil [8,10], which promote the
Asiatic pennywort or Indian pennywort, is a proliferation and repair of skin cells, as well as its
common herbaceous plant widely distributed in anti-inflammatory and antioxidant properties that
Asia, Africa and Latin America. It has been used alleviate inflammation at the wound site.
in traditional herbal medicine to treat various
diseases and symptoms, and has gained Additionally, this study delves into the
extensive study interest in recent years, mechanisms of action of Gotu kola oil. First, Gotu
particularly in the context of skin health and kola oil accelerates the repair of damaged tissue
wound healing [5,6]. Gotu kola oil, at room by promoting cell proliferation and collagen
temperature, is a dark blue viscous liquid, also synthesis at the wound site [8,11]. Growth factors
known as Gotu kola extract. Its chemical name is are pivotal in orchestrating the intricate process
madecassoside, with a molecular formula of of wound healing. Among them, TGF-β1 plays a
C15H18. It is a widely used topical medication crucial role and exerts its influence throughout
commonly employed for treating burns and the various stages of wound healing via the
frostbite in everyday life. Gotu kola oil contains smad2/3 signaling pathway [12,13]. In addition,
various bioactive compounds, including TGF-β1 not only governs the proliferation of
triterpenoids and polysaccharides, which exhibit keratinocytes, facilitating epidermal regeneration,
significant anti-inflammatory properties. These but also regulates fibroblast proliferation, thereby
compounds mitigate inflammatory responses by promoting collagen synthesis and the formation
inhibiting the production of inflammatory of granulation tissue. These mechanisms are
mediators and the activation of inflammatory believed to contribute to the wound healing-
signaling pathways, helping to alleviate skin promoting effects observed in the context of Gotu
inflammation and discomfort [7]. Gotu kola oil kola oil. Furthermore, the anti-inflammatory effect
contains polyphenolic compounds such as of Gotu kola oil reduces inflammation at the
catechins and anthocyanins, which possess wound site by inhibiting the release of
potent antioxidant properties. These antioxidants inflammatory mediators, contributing to the
help neutralize free radicals, reduce oxidative healing process. It also reduces the adverse
stress, and consequently decrease cellular and effects of oxidative stress on wound healing
tissue damage. Gotu kola oil has been shown to through antioxidant mechanisms [14,15]. These

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 Xu et al

mechanisms interact synergistically to accelerate Firstly, it is important to note that this study
the rapid healing of skin wounds. primarily focused on a laboratory mouse model
and further studies should be conducted in
Furthermore, it is widely acknowledged that settings that closely resemble clinical conditions.
vascular regeneration plays a pivotal role in the Secondly, additional investigations are required
process of wound repair [12]. Vascular to determine the optimal usage and dosage of
regeneration not only facilitates the Gotu kola oil for the most effective treatment
establishment of new tissue structures but also regimen. Lastly, a more profound elucidation of
provides essential materials and nutrients for the molecular mechanisms underlying the effect
repair while aiding in waste removal. Notably, of Gotu kola oil is necessary to identify additional
growth factors such as HGF and VEGF assume pathways and potential targets.
crucial roles in promoting the formation of blood
vessels, significantly augmenting the population Previous studies suggested that its notable anti-
of endothelial progenitor cells, facilitating their inflammatory effect is achieved through the
mobilization and differentiation, and thereby inhibition of inflammatory factor release and the
facilitating vascular regeneration. Additionally, regulation of immune cell activity, which
VEGF also exerts a significant impact on contributes to reducing inflammation at the
vascular permeability and anti-apoptotic wound site and expediting the healing process.
mechanisms, thus promoting the formation of However, this particular study did not specifically
new blood vessels [12,13]. address this aspect. Therefore, future study
could consider expanding the sample size,
In the present study, immunohistochemical conducting large-scale multicenter clinical
labeling utilizing CD34 antibodies was employed studies to validate the findings of this study, and
to identify newly formed blood vessels, comprehensively evaluate the practical
characterized by brown circular or strip-like application value of Gotu kola oil.
structures, followed by subsequent counting and
comparative analysis. On the 3rd and 7th day, CONCLUSION
study group exhibited higher levels of VEGF
expression compared to control group, which Gotu kola oil shows great potentials for skin
aligns with the observed trend of microvessel wound repair. A comprehensive understanding of
density changes. Although the differences were the healing bioactivity and mechanisms of action
not significant, this may be due to the uneven of Gotu kola oil has been achieved, providing
distribution of identified vascular markers. evidence for the advancement of more
efficacious strategies in skin wound repair.
Gotu kola oil may promote the generation of Subsequent investigations will continue to
capillaries at the wound site. However, on the broaden this field, aiming to ultimate advance to
14th day, the microvessel density in the study achieve patient recovery and improved overall
group was lower than that in the control group. quality of life.
This may be because the wound healing rate in
the study group was faster, entering the stage of DECLARATIONS
granulation tissue filling the wound earlier, at
which point capillaries cease to regenerate. In Acknowledgements
contrast, the wound healing rate in control group
was slower and was still in the phase of rapid None provided.
new blood vessel formation. The disparity in
microvessel density could potentially be
Funding
attributed to varying stages of the healing
process. Gotu kola oil which comprises a diverse
range of bioactive constituents such as None provided.
polysaccharides, alkaloids, and polyphenols, has
demonstrated multiple bioactivities in laboratory Ethical approval
investigations. These activities encompass anti-
inflammatory, antioxidant, antimicrobial, and cell None provided.
proliferation-stimulating properties [16,17].
Availability of data and materials
Study limitations
The datasets used and/or analyzed during the
Although this study provides evidence to support current study are available from the
the potential application of Gotu kola oil in skin corresponding author on reasonable request.
wound repair, there are still some limitations.

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 Xu et al

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T. A systematic review of the effect of Centella asiatica
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19(6): 3266.
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