AUTHOR=Sari Dwi Cahyani Ratna , Budiharjo Santosa , Afifah Husnari , Jasmin Destantry , Kokasih Orisativa , Putri Tiara Gitami , Arifiani Karina , Setyaningsih Wiwit Ananda Wahyu , Arfian Nur 

TITLE=Centella asiatica Extract Attenuates Kidney Fibrosis Through Reducing Mesenchymal Transition and Inflammation in Ureteral Ligation Model in Mice

JOURNAL=Frontiers in Pharmacology

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2021.621894

DOI=10.3389/fphar.2021.621894

ISSN=1663-9812

ABSTRACT=<p><bold>Background:</bold> Kidney fibrosis is the common final pathway of chronic kidney disease (CKD)<italic>,</italic> and is characterized by inflammation, mesenchymal transition with myofibroblast formation and epithelial to mesenchymal transition (EMT). <italic>Centella asiatia</italic> (CeA) is an herb that has a reno-protective effect. However, its mechanism of action in kidney fibrosis has not been elucidated.</p><p><bold>Aim:</bold> To elucidate the effect of CeA in amelioration of kidney fibrosis in a unilateral ureteral obstruction (UUO) model and focus on mesenchymal transition and inflammation.</p><p><bold>Methods:</bold> Unilateral ureteral obstruction was performed in male Swiss-background mice (age: 2–3 months, weight: 30–40 g, UUO group <italic>n</italic> = 6) to induce kidney fibrosis. Two doses of CeA extract with oral administration, 210 and 840 mg/kg body weight were added in UUO (U+C210 and U+C840 groups, each <italic>n</italic> = 6). The sham operation procedure was performed for the control group (SO, <italic>n</italic> = 6). The mice were euthanized at day-14 after operation. Tubular injury and interstitial fibrosis area fractions in kidney tissues of the mice were quantified based on periodic acid-Schiff (PAS) and Sirius Red (SR) staining. Immunostaining was performed for examination of fibroblast (PDGFR-β), myofibroblast (α-SMA), Monocyte Chemoattractant Protein-1 (MCP-1) and macrophage (CD68), meanwhile double immunofluorescence was performed with PDGFR-β and α-SMA. Reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to examine mRNA expression of TGF-β, Collagen-1, Snail, E-cadherin, vimentin, fibroblast-specific protein 1 (FSP-1), CD68, toll-like receptor 4 (TLR4), and MCP-1.</p><p><bold>Results:</bold> We observed a significantly higher interstitial fibrosis area fraction and tubular injury (<italic>p</italic> &lt; 0.001) with fibroblast expansion and myofibroblast formation in the UUO group than in the SO group. These findings were associated with higher mRNA expression of TGF-β, Collagen-1, Snail, vimentin, FSP-1, CD68, TLR4, and MCP-1 and lower mRNA expression of E-cadherin. The U+C840 group had a significantly lower tubular injury score and interstitial fibrosis area fraction, which associated with downregulation of mRNA expression of TGF-β, Collagen-1, Snail, vimentin, FSP-1, CD68, TLR4, and MCP-1, with upregulation of mRNA expression of E-cadherin. Immunostaining observation revealed the U+C840 group demonstrated reduction of macrophage infiltration and myofibroblast expansion.</p><p><bold>Conclusion:</bold> CeA treatment with dose-dependently ameliorates mesenchymal transition and inflammation in kidney fibrosis in mice.</p>