Andrographis paniculata Leaves Extract Inhibit TNF-α and Caspase-3 Expression of Septic Rats’ Intestinal Tissues

Ryco Giftyan Ardika, Bernardus Parish Budiono, Nyoman Suci Widiastiti, Nani Maharani, Neni Susilaningsih, Ferry Sandra


BACKGROUND: Microcirculation and cellular disturbances caused by sepsis might trigger significant intestinal damage. Andrographis paniculata extract decreases inflammatory intestinal epithelial cells with its role as an antiparasitic and anti-inflammatory agent. However, A. paniculata extract’s effect on sepsis have not been commonly studied, especially in the intestinal tissues. Therefore, this study was conducted to determine A. paniculata leaves extract (APLE) effect in sepsis-induced intestinal tissues of rats by examining the expression of inflammatory cytokines involved in sepsis, namely tumor necrosis factor (TNF)-α and Caspase-3.

METHODS: Rats were divided into five groups; two groups received no pretreatment and the other three groups received 200, 400, and 500 mg/kg BW/day APLE, respectively. Three pretreated groups and one group with no pretreatment were then injected with 1 mg/200 g BW lipopolysaccharides (LPS) intraperitoneally to create septic rat models. Three days after the LPS-induction, rats were euthanized and the expression of TNF-α and Caspase-3 were assessed based on the immunohistochemical staining of rats’ intestinal tissues.

RESULTS: Compared with NaCl (sham), LPS significantly (p<0.001) induced TNF-α expression from 6.60±1.36 to 25.37±1.74. Pretreatment of 200, 400, and 500 mg/kg BW/day APLE could significantly (p<0.001) inhibit the LPS-induced TNF-α expression (18.82±1.36, 11.45±1.18, and 6.89±1.90, respectively). Similar with TNF-α, compared with NaCl (sham), LPS significantly (p<0.001) induced Caspase-3 expression from 6.92±1.66 to 23.59±2.25. Pretreatment of 200, 400, and 500 mg/kg BW/day APLE could significantly (p<0.001) inhibit the LPS-induced Caspase-3 expression (17.47±1.68, 12.99±1.51, and 5.59±1.51, respectively).

CONCLUSION: The pretreatment of APLE could inhibit the LPS-induced TNF-α and Caspase-3 expression, therefore APLE could be suggested as a potential sepsis-preventing agent.

KEYWORDS: Andrographis paniculata, sepsis, TNF-α, Caspase-3, lipopolysaccharide


Full Text:



World Health Organization. Global Report on the Epidemiology and Burden of Sepsis: Current Evidence, Identifying Gaps and Future Directions. Geneva: World Health Organization; 2020, article.

Yealy DM, Mohr NM, Shapiro NI, Venkatesh A, Jones AE, Self WH. Early care of adults with suspected sepsis in the emergency department and out-of-hospital environment: A consensus-based task force report. Ann Emerg Med. 2021; 78(1): 1–19, CrossRef.

Kevin M, Widyastiti NS, Budijitno S, Prajoko YW, Susilaningsih N. The protective effect of Andrographis paniculata against lipopolysaccharide-induced sepsis in lung tissues of a rat model through the decrease of ICAM-1 and E-selectin expression. Indones Biomed J. 2023; 15(6): 411–9, CrossRef.

Cecconi M, Evans L, Levy M, Rhodes A. Sepsis and septic shock. Lancet. 2018; 392(10141): 75–87, CrossRef.

Al-Sadi R, Guo S, Ye D, Rawat M, Ma TY. TNF-α modulation of intestinal tight junction permeability is mediated by NIK/IKK-α axis activation of the canonical NF-κB pathway. Am J Pathol. 2016; 186(5): 1151–65, CrossRef.

Alvarez S, Blanco A, Fresno M, Muñoz-Fernández MÁ. TNF-α contributes to caspase-3 independent apoptosis in neuroblastoma cells: Role of NFAT. PLoS One. 2011; 6(1): e16100, CrossRef.

Freedman JC, Navarro MA, Morrell E, Beingesser J, Shrestha A, McClane BA, et al. Evidence that Clostridium perfringens enterotoxin-induced intestinal damage and enterotoxemic death in mice can occur independently of intestinal caspase-3 activation. Infect Immun. 2018; 86(7): e00931–17, CrossRef.

Billmeier U, Dieterich W, Neurath MF, Atreya R. Molecular mechanism of action of anti-tumor necrosis factor antibodies in inflammatory bowel diseases. World J Gastroenterol. 2016; 22(42): 9300–13, CrossRef.

Marshall JC, Leligdowicz A. Gaps and opportunities in sepsis translational research. eBioMedicine. 2022; 86: 104387, CrossRef.

Sandra F. Role of herbal extract in stem cell development. Mol Cell Biomed Sci. 2018; 2(1): 19–22, CrossRef.

Qurrotuaini SP, Wiqoyah N, Mustika A. Antimicrobial activity of ethanol extract of Centella asiatica leaves on Proteus mirabilis, Proteus vulgaris, and Yersinia enterocolitica in vitro. Mol Cell Biomed Sci. 2022; 6(3): 135–40, CrossRef.

Suastika LOS, Oktaviono YH, Soemantri D, Sandra F. Purple sweet potato extract and vitamin C increase the proliferation of endothelial progenitor cells from stable coronary artery disease patients. Bali Med J. 2021; 10(1): 243–8, CrossRef.

Andari D, Khan FI, Jakfar SI. Methanol extract of katuk (Sauropus androgynus) leaves as an anti-inflammatory agent: Animal study in carrageenan-induced rat models of inflammation. Mol Cell Biomed Sci. 2022; 6(3): 129–34, CrossRef.

Ranggaini D, Sandra F, Halim J, Ichwan SJA, Djamil MS. Curcuma xanthorrhiza rhizome extract induces apoptosis in HONE-1 nasopharyngeal cancer cells through bid. Indones Biomed J. 2023; 15(1): 100–5, CrossRef.

Sikumalay A, Suharti N, Masri M. Efek antibakteri dari rebusan daun sambiloto (Andrographis paniculata Nees) dan produk herbal sambiloto terhadap Staphylococcus aureus. J Kesehat Andalas. 2014; 5(1): 196–200, CrossRef.

Lin HC, Li CC, Yang YC, Chiu TH, Liu KL, Lii CK, et al. Andrographis paniculata diterpenoids and ethanolic extract inhibit TNFα-induced ICAM-1 expression in EA.hy926 cells. Phytomedicine. 2019; 52: 157–67, CrossRef.

Astuti NT, Novitasari PR, Tjandrawinata R, Nugroho AE, Pramono S. Anti‐diabetic effect of andrographolide from Sambiloto herbs (Andrographis paniculata (Burm.f.) Nees) through the expression of PPARγ and GLUT‐4 in adipocytes. Indones J Biotechnol. 2022; 27(4): 203–11, CrossRef.

Yu L, Liu Y, Cao C, Yang L, Liu H, Wang C. Andrographolide attenuates inflammation due to intra-abdominal sepsis by enhancing bacterial clearance in mice. J Inflamm Res. 2023; 16: 4413–23, CrossRef.

Kusmardi K, Ariffandi B, Lubis NS, Lestari TW, Intan PR, Pakpakan A. Effects of sambiloto (Andrographis paniculata) extract and spirulina (Spirulina platensis) administration on Ki-67 protein expression in the colon epithelial cells of Plasmodium berghei-infected mice. Indones Biomed J. 2022; 14(1): 84–90, CrossRef.

Wanandi SI, Syahrani RA, Suraduhita A, Yunita E. Andrographolide reverses doxorubicin resistance in human breast cancer stem cells by regulating apoptotic gene expressions. Indones Biomed J. 2023; 15(5): 288–96, CrossRef.

Pourdast A, Sanaei M, Jafari S, Mohammadi M, Khalili H, Shafiee G, et al. Effect of SeptimebTM as a new natural extract on severe sepsis: A randomized clinical trial. Caspian J Intern Med. 2017; 8(1): 35–43, PMID.

Lee YY, Yuk HJ, Saba E, Kim SD, Kim DS, Kopalli SR, et al. Duchesnea indica extract ameliorates LPS-induced septic shock in mice. Evid Based Complement Alternat Med. 2022: 2022: 5783867, CrossRef.

Williams JM, Duckworth CA, Watson AJM, Frey MR, Miguel JC, Burkitt MD, et al. A mouse model of pathological small intestinal epithelial cell apoptosis and shedding induced by systemic administration of lipopolysaccharide. Dis Model Mech. 2013; 6(6): 1388–99, CrossRef.

Kuzmich NN, Sivak KV, Chubarev VN, Porozov YB, Savateeva-Lyubimova TN, Peri F. TLR4 signaling pathway modulators as potential therapeutics in inflammation and sepsis. Vaccines. 2017; 5(4): 34, CrossRef.

Brentnall M, Rodriguez-Menocal L, De Guevara RL, Cepero E, Boise LH. Caspase-9, caspase-3 and caspase-7 have distinct roles during intrinsic apoptosis. BMC Cell Biol. 2013; 14(1): 32, CrossRef.

Lin BF, Chao WW, Kuo YH, Hsieh SL. Inhibitory effects of ethyl acetate extract of Andrographis paniculata on NF-κB trans-activation activity and LPS-induced acute inflammation in mice. Evid Based Complement Altern Med. 2011; 2011: 254531, CrossRef.

Guo W, Liu W, Chen G, Hong S, Qian C, Xie N, et al. Water-soluble andrographolide sulfonate exerts anti-sepsis action in mice through down-regulating p38 MAPK, STAT3 and NF-κB pathways. Int Immunopharmacol. 2012; 14(4): 613–9, CrossRef.

Low M, Khoo CS, Münch G, Govindaraghavan S, Sucher NJ. An in vitro study of anti-inflammatory activity of standardised Andrographis paniculata extracts and pure andrographolide. BMC Complement Altern Med. 2015; 15: 18, CrossRef.


Copyright (c) 2024 The Prodia Education and Research Institute

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.


Indexed by:






The Prodia Education and Research Institute