1.Upregulation of p16, Bax and Bcl-2 mRNA Expression Associated with Epithelial Apoptosis and Myofibroblast Proliferation in Kidney Fibrosis Model in Mice
Ike Sulistiyowati1 ; Junaedy Yunus ; Dwi Cahyani Ratna Sari ; Nur Arfian
Malaysian Journal of Medical Sciences 2020;27(2):37-44
Background: Cellular senescence may play a role in the development of kidney fibrosis,
but its specific association with apoptosis or proliferation have yet to be determined.
Objectives: This study aims to determine the effects of unilateral ureteral obstruction
(UUO) on proliferation, cellular senescence and apoptosis in kidney fibrosis.
Methods: A unilateral ureteral obstruction (UUO) procedure was performed to induce
kidney fibrosis in 24 Swiss mice (3 months old, 30 g–40 g). Mice were sacrificed on day 3 (UUO3,
n = 6), day 7 (UUO7, n = 6) and day 14 (UUO14, n = 6). Sham operation (SO) procedures were
performed on the control group. The expression of Bcl-2, p16 and Bax mRNA was quantified with
reverse transcription polymerase chain reaction (RT-PCR). Immunohistochemical (IHC) staining
with anti-Bcl-2 and p53 antibodies was used to determine the localisation of proliferation and
apoptosis. Data were analysed using one-way ANOVA followed by a post hoc least significant
difference (LSD) test (P < 0.05)
Results: RT-PCR analysis showed higher mRNA expression of Bcl-2, p16 and Bax in
the UUO groups compared with SO group (P < 0.05). Immunostaining showed that Bcl-2 and
p53 expression in tubular epithelium in the UUO groups, except Bcl-2 expression was found in
interstitial areas of UUO14 group.
Conclusion: Senescence in UUO might be associated with epithelial apoptosis and
myofibroblast proliferation.
2.The application of augmented reality for improving clinical skills: a scoping review
Mahmud MAHMUD ; Dwi Cahyani Ratna SARI ; Djayanti SAR ; Nur ARFIAN ; Muhammad Ary ZUCHA
Korean Journal of Medical Education 2024;36(1):65-79
Augmented reality technology had developed rapidly in recent years and had been applied in many fields, including medical education. Augmented reality had potential to improve students’ knowledge and skills in medical education. This scoping review primarily aims to further elaborate the current studies on the implementation of augmented reality in advancing clinical skills. This study was conducted by utilizing electronic databases such as PubMed, Embase, and Web of Science in June 2022 for articles focusing on the use of augmented reality for improving clinical skills. The Rayyan website was used to screen the articles that met the inclusion criteria, which was the application of augmented reality as a learning method in medical education. Total of 37 articles met the inclusion criteria. These publications suggested that using augmented reality could improve clinical skills. The most researched topics explored were laparoscopic surgery skills and ophthalmology were the most studied topic. The research methods applied in the articles fall into two main categories: randomized control trial (RCT) (29.3%) and non-RCT (70.3%). Augmented reality has the potential to be integrated in medical education, particularly to boost clinical studies. Due to limited databases, however, any further studies on the implementation of augmented reality as a method to enhance skills in medical education need to be conducted.
3.Imbalance between Endothelin-1 and eNOS Expression Associates with Tubular Injury in Mice with 5/6 Subtotal Nephrectomy
Muhammad Mansyur Romi ; Nur Arfian ; A.A.N Nata Baskara ; Dwi Cahyani Ratna Sari
Malaysian Journal of Medicine and Health Sciences 2020;16(Supp 3,June):82-85
Introduction: Chronic Kidney Diseases (CKD) leads to kidney fibrosis which characterized by tubular injury and atrophy with interstitial fibrosis. Endothelin-1 (ET-1) and endothelial Nitrite Oxide Synthase (eNOS) are known to play
role in CKD and kidney fibrosis, although their correlation with tubulo-interstitial injury have not been understood
clearly. Methods: 5/6 Subtotal Nephrectomy (SN) was performed in male Swiss Background mice to induce CKD.
Sham operation (SO, n=5) procedure was performed on mice as control. The mice were sacrificed in day 7 (1N, n=5)
and day 28 (4N, n=5) after operation. We measured creatinine serum to assess renal function. Tubular injury score
was quantified based on Periodic-Acid Schiff (PAS) staining. Prepro-ET-1 and eNOS were quantified using RT-PCR.
Results: SN_1N and SN_4N groups had significant higher of serum creatinine and tubular injury from SO group. Densitometry analysis of RT-PCR revealed up-regulation of prepro-ET-1 mRNA expression in SN_1N and SN_4N (p<0.05
vs SO). Meanwhile, we found a significant increase of eNOS expression in SN_1N, and then it reduced significantly
in SN_4N. We found significant parallel correlation between ET-1 and tubular injury expression (r: 0.768;p<0,05),
meanwhile there were insignificant inverse correlation between eNOS and tubular injury (r: -0.354;p>0.05). Conclusion: eNOS might play role as a counterbalance in the up regulation of ET-1 in acute condition after SN. However, it
failed in chronic condition. These lead to deterioration of renal function and tubular injury. An imbalance between
ET-1 and eNOS expression in chronic CKD model might play role in profound renal damage.
4.Ethanol Extract of Centella asiatica (Gotu Kola) Attenuates Tubular Injury Through Inhibition of Inflammatory Cytokines and Enhancement of Anti-Fibrotic Factor in Mice with 5/6 Subtotal Nephrectomy
Nur Arfian ; Wiwit Ananda Wahyu Setyaningsih ; Nungki Anggorowati ; Muhammad Mansyur Romi ; Dwi Cahyani Ratna Sari
Malaysian Journal of Medical Sciences 2019;26(5):53-63
Background: Chronic kidney disease (CKD) leads to inflammation, fibrosis and
destruction of the renal architecture. Centella asiatica (CeA) is an herbaceous plant with antiinflammatory
effects. We aimed to elucidate the effect of CeA on inflammation, fibrosis, vascular
remodelling and antifibrotic substances in a 5/6 subtotal nephrectomy (SN) model in mice.
Methods: Mice were divided into three groups: sham operation (SO, n = 6), 5/6 SN for
seven days (SN7, n = 7) and SN7 with oral CeA treatment (SN7-CeA, n = 7). At day 7, mice were
euthanised, kidneys were harvested and stained with periodic-acid Schiff (for tubular injury
and glomerulosclerosis) and sirius red (for fibrosis and vascular remodeling) staining. mRNA
expression of prepro-endothelin-1, nephrin, E-cadherin, bone morphogenic protein-7 (BMP-7),
toll-like receptor 4 (TLR4), tumour necrosis factor-α (TNFα) and hepatocyte growth factor (HGF)
were quantified using reverse transcriptase-PCR.
Results: SN group demonstrated significant higher interstitial fibrosis, vascular
remodeling, tubular injury and glomerulosclerosis (P < 0.01) compared to SO group. Meanwhile,
in SN7-CeA demonstrated attenuation of vascular remodeling as shown by significant higher lumen
area with lower Wall/Lumen area ratio compared to SN7. RT-PCR analysis showed up-regulation
of nephrin, BMP-7 and E-cadherin mRNA expression (P < 0.05) and down-regulation of ppET-1 in
SN7-CeA group compared to SN7 group (P < 0.05).
Conclusion: CeA may ameliorate renal injury in the SN model in mice.
5.Effect of High-Fat Diet on SOD2, GPx, NeuN and BDNF Expression on Frontal Lobe of Obese Rats
Mohammad Ariq Nazar ; Dwi Cahyani Ratna Sari ; Agung Putra ; Nur Arfian
Malaysian Journal of Medicine and Health Sciences 2021;17(No.4):162-165
Introduction: Obesity has been demonstrated to induce oxidative stress and inflammation processes that lead to
senescence in brain cells. Obesity-induced cellular senescence in the brain is still widely investigated. This study
aimed to investigate the expression of antioxidant and neuronal markers in the frontal lobes of obese rats. Methods:
Eighteen adult rat Sprague Dawley divided into three groups: Control (SO), Obese-2 (DIO2), and Obese-4 (DIO4)
were observed. Control rats were fed with a standard diet AIN 76A for two month. In contrast, DIO2 and DIO4 rats
were fed with a high-fat diet daily for two and four months, respectively. After being sacrificed, the rats’ brains were
dissected out then the frontal lobes were used for RNA extraction. Reverse transcriptase PCR of SOD2, GPx, BDNF,
NeuN and beta-actin was performed to investigate the relative expression of the antioxidant and neuronal markers.
Results: DIO2 and DIO4 groups had significantly increased body Weights, blood glucose level and triglyceride level
after being fed with a high-fat diet for two and four months, respectively. The DIO4 group had the significantly lowest
mRNA expressions of SOD2, GPx, BDNF and NeuN. Conclusion: Decreased antioxidant and neuronal markers in
the rats frontal lobes were observed as the chronic effect of obesity.
6.Uric Acid Induces Inflammation, Hepatocyte Apoptosis and Deterioration of Liver Function
Dwi Cahyani Ratna Sari ; Rina Nofrienis ; Muhammad Mansyur Romi ; Untung Tranggono ; Eryna Ayu Nugra Desita ; Nur Arfian
Malaysian Journal of Medicine and Health Sciences 2020;16(Supp 3,June):49-55
Introduction: Uric acid is a common cause of liver tissue damage due to its hepatotoxic effect. This study is aimed to
investigate: (1) the effect of uric acid on liver damage which can be seen from the serum levels of SGOT and SGPT,
(2) the inflammatory response demonstrated by TLR-4 and MCP-1 mRNA expression, and (3) the proportion of hepatocytes apoptosis in mice. Methods: A total of 25 adult male Swiss-Webster mice were divided into five groups:
one control group and four uric acid groups (AU7, AU14, AU21 and AU28). The uric acid groups were administered
with 125 mg/kgBW uric acid for 7, 14, 21, and 28 days. Following the treatment, mice were terminated and the liver
was harvested. Blood sample was taken from retro-orbital vein to assess serum uric acid, SGOT, and SGPT levels.
RT-PCR was performed to examine the mRNA expressions of TLR-4 and MCP-1. TUNEL staining was used to assess
the proportion of apoptotic hepatocytes. Results: Induction of uric acid caused hyperuricemia, increased expression
of TLR-4 and MCP-1 mRNA significantly (p<0.05) which indicated an inflammatory reaction. The levels of SGOT
and SGPT were elevated significantly (p<0.05), as well as the number of hepatocyte apoptosis (p<0.05). Conclusion: Hyperuricemia affected the inflammatory response by increasing the mRNA expression of TLR-4 and MCP-1.
An increased number of apoptotic hepatocytes was likely caused by the ongoing inflammatory reaction during the
induction of uric acid.
7.Expression of Senescence Markers Increased in the Frontal Lobes of Obese Rats
Dwi Cahyani Ratna Sari ; Mawaddah Ar Rochmah ; Wiwit Ananda Wahyu Setyaningsih ; Josephine Debora ; Dhite Bayu Nugroho ; Nur Arfian
Malaysian Journal of Medicine and Health Sciences 2020;16(Supp 3,June):62-66
Introduction: Obesity has been demonstrated to induce oxidative stress and inflammation processes that lead to
senescence. Obesity-induced cellular senescence in the brain is still widely investigated. This study aimed to investigate the expression of senescence markers in the frontal lobes of obese rats. Methods: Three groups of rats: control,
Obese-2 (Ob-2), and Obese-4 (Ob-4) were observed. Control rats were fed with a standard diet for one month. In
contrast, Ob-2 and Ob-4 rats were fed with a high-fat diet daily for two and four months, respectively. After being
sacrificed, the rats’ brains were dissected out then the frontal lobes were used for RNA extraction. Reverse transcriptase PCR of p-16, p-21, and beta-actin was performed to investigate the relative expression of the senescence
markers. Results: Ob-2 and Ob-4 groups had significantly increased body weight after being fed with a high-fat diet
for two and four months, respectively. The mRNA expressions of p-16 and p-21 in the frontal lobes of three groups
showed similar patterns. The ob-4 group had the highest mRNA expressions of both p-16 and p-21. In comparison
to control and Ob-2 groups, the mRNA expressions of p-16 and p-21 were markedly increased. Furthermore, the
mRNA expressions of p-16 and p-21 between control and Ob-2 groups were comparable. Conclusion: Increased
senescence markers in the rats’ frontal lobes were observed as the chronic effect of obesity.
8.Ethanol Extract of Yacon Leaves (Smallanthus sonchifolius) Attenuates Fibroblast and Myofibroblast Expansion in Association With Downregulation of TGF β1 and Snail mRNA Expression in 5/6-Subtotal Nephrectomy Model in Mice
Ermawati ; Nur Arfian ; Setyo Purwono ; Muhammad Mansyur Romi ; Dwi Cahyani Ratna Sari
Malaysian Journal of Medicine and Health Sciences 2022;18(No.3):85-91
Introduction: Chronic kidney disease (CKD) is characterized by fibroblast activation, myofibroblast formation, and
up-regulation of transforming growth factor-β1 (TGF-β1) that may activate Snail in fibroblast to myofibroblast transition. Ethanol extract of Yacon leaves is known to have a renoprotective effect on diabetic nephropathy but its effect
in the CKD model is unknown. This experimental study aimed to elucidate the effect of ethanol extract from Yacon
leaves in attenuating renal failure in a CKD mice model. Methods: Male Swiss-Webster mice (3 months, 30–40
grams, n=25) underwent 5/6 subtotal nephrectomy (SN) to induce CKD. The mice were divided into five groups: SN,
SN mice with oral treatment of Yacon leaves ethanol extract with doses 0.735 μg/kg (SN+YK1), 1.47 μg/kg (SN+YK2),
and 2.94 μg/kg (SN+YK3), and a Sham operation (SO) group with aquadest 0.1% supplementation. Mice were euthanized on day 14 after the operation and kidneys were harvested. Paraffin sections were used for histological analysis.
Immunostaining was done for quantifying fibroblasts and myofibroblasts. We performed RT-PCR to measure TGF-β1
and Snail mRNA expressions. Results: The SN group had significantly higher fibroblast number, myofibroblast fraction area, TGF-β1 and Snail mRNA expressions compared to the SO. The fibroblasts number (p<0.001) and myofibroblast fraction areas (p<0.001) were significantly lower in Yacon treated-groups compared to the SN group. RT-PCR
analysis showed lower mRNA expressions of TGF-β1 and Snail, but no significant differences were found among the
various Yacon treated-groups. Conclusion: Ethanol extracts of Yacon leaves improved kidney damage in male mice
with 5/6 subtotal nephrectomy model.
9.Ethanolic Extract of Centella asiatica Ameliorates Kidney Ischemia/reperfusion Injury Through Inhibition of Inflammatory Process
Nur Arfian ; Arindira Maharani ; Elida Fadhilatul Latifa ; Indhah Kusumaningtyas ; Monica Agnes Witono ; Kemala Athollah ; Ratih Kemalasari ; Gita Mumtarin Dara ; Wiwit Ananda Wahyu Setyaningsih ; Dwi Cahyani Ratna Sari
Malaysian Journal of Medicine and Health Sciences 2020;16(Supp 3,June):71-77
Introduction: Kidney ischemic-reperfusion injury (IRI) is the main cause of acute kidney injury (AKI) which leads to
the inflammation epithelial apoptosis and interstitial fibrosis as the chronic consequenses. Centella asiatica (CeA)
has been known to have various pharmacological effects such as, anti-inflammatory, antioxidant, anti-fibrosis, and,
anti-apoptosis. We aimed to elucidate the role of CeA in inhibiting kidney injury and infammatory mediators due to
kidney IRI. Methods: Kidney IRI were performed with bilateral renal pedicles clamping in Swiss background mice (3
months-old, 30-40 grams) for 30 minutes (IR group, n=6), then terminated at day 7 after operation. At the next day,
the mice that have been underwent bilateral kidney IRI were administered per-orally with ethanolic extract of CeA
(210 mg/kg of BW, CeA1 group, n=6, and 420 mg/kg of BW, CeA2 group, n=6). The Sham Operation (SO group,
n=6) was used as control. At the day 7 after the surgery, the mice were sacrificed and the kidneys were harvested.
The kidney was used to assess tubular injury, interstitial fibrosis, and macrophage number, and another kidney was
used to assess the mRNA expression of TLR4. Data were quantified using SPSS 22. Results: Kidney IRI produced significantly higher tubular injury, interstitial fibrosis and macrophage number (p<0.05) compared to SO with upregulating TLR4 mRNA expression (p<0.05). CeA treatment attenuated the tubular injury, interstitial fibrosis, macrophage
number, and TLR4 mRNA expression which obviously shown in higher-dose of CeA (p<0.05). Conclusion: CeA
ameliorates tubular injury, kidney fibrosis, and inflammatory mediators due to kidney IRI.
10.Upregulation of ppET-1/ETBR/eNOS mRNA Expression After Calcitriol Treatment in Chronic Kidney Diseases Model in Rats
Anak Agung Ngurah Nata Baskara ; Nur Arfian ; Wiwit Ananda Wahyu Setyaningsih ; Muhammad Mansyur Romi ; Dwi Cahyani Ratna Sari
Malaysian Journal of Medicine and Health Sciences 2022;18(No.5):23-29
Introduction: Myofibroblast formation in the interstitial area is the hallmark of chronic kidney disease (CKD). Endothelin signalling has been known to play role in physiology and pathophysiology in the kidney. Vitamin D has a
reno-protective effect through inhibiting inflammation and fibrosis. However, the interaction between vitamin D and
endothelin signalling in the CKD model has not been elucidated yet. Therefore, we aimed to check the difference
impact of endothelin (ET) receptor in CKD. Methods: Sprague Dawley rats (3-months-old, 150-250grams) underwent
5/6 subtotal nephrectomy (SN) to induce CKD. Then, it was divided into 4 groups (each contains 6 rats): sham operation (SO), 5/6 subtotal nephrectomy (SN), calcitriol groups (0.01µg/100grBW/day (SN-D1), and 0.05µg/100grBW/day
(SN-D2). Calcitriol was administered for 14 days after the surgery. The Sham Operation (SO) group was injected with
NaCl. At the specified date, the rats were sacrificed and the kidneys were harvested. Fibrosis was quantified based
on Sirius Red staining. Immunostaining was done for localizing fibroblast (PDGFRβ). The mRNA expressions of prepro-ET-1, endothelin receptor A (ETAR), endothelin receptor B (ETBR), and endothelial nitrite oxide synthase (eNOS)
were quantified using reverse-transcriptase PCR (RT-PCR). Results: The CKD promotes an elevation of prepro-ET-1,
ETBR, and eNOS, and reduction of ETAR (p<0.05) mRNA expression compared to the SO group. Administration of
calcitriol (SN-D1 and SN-D2) showed the vice versa effects. However, only SN-D2 group consistently showed statistically significant differences whenever compared to either SO or SN groups. Conclusion: Calcitriol might attenuate
interstitial fibrosis in CKD model via ET-1/eNOS signalling.