It is postulated that beneficial effects of A. Andrographolide is also well known for its broad-spectrum antiviral properties. This finding opens the possibility to develop A. In comparison to our previous study, 30 A.
This is an additional rationale to support A. Since both IFA and plaque assays investigate the distinct steps in the SARS-CoV-2 life cycle the expression of viral protein vs the production of infectious progenies , interpretation of IC 50 values of the compound obtained from such assays could be made differently.
This depends on the potency of the extract or compound in perturbing a specific step of the SARS-CoV-2 life cycle, either the viral protein expression or the particle release.
On the other hand, the IC 50 value from the plaque assay could be indicated as the half-maximal concentration of the compound able to inhibit infectious virion production. From our data, the IC 50 values measured by the plaque assay were lower than those of the high-content imaging IFA.
This can be interpreted as both A. The late phases of the SARS-CoV-2 life cycle include viral assembly and maturation, transportation along the secretory pathway, or particle release. Then the inoculum was removed, and the cells were washed before adding fresh medium without the compound into the cells.
This study was associated with several limitations. Although our findings support andrographolide as a promising candidate for further anti-SARS-CoV-2 development, the low bioavailability of andrographolide might pose a limitation for clinical applications. The safety and efficacy of andrographolide should be further investigated in preclinical animal models and clinical studies.
Potent anti-SAR-CoV-2 activities, together with the favorable cytotoxicity profiles, support further development of A. An immortalized hepatocyte-like cell line imHC was established in-house as previously described, 57 and its hepatic phenotypes were characterized previously.
The plant was identified, authenticated by Chao Phya Abhaibhubejhr Hospital, Prachin Buri, Thailand, and deposited at the herbarium unit. The powder of A. After 24 h, the liquid fraction was separated using a thin straining cloth, then filtered through filter paper by a vacuum pump. The andrographolide content in the crude extract was 7. Then, culture medium was discarded and washed once with phosphate-buffered saline PBS. Each concentration of drugs, crude extract, or active compound was inoculated into the culture medium.
The experiment was performed in triplicate. The fixed cells were washed once with PBS with 0. After incubation, cells were washed with PBST three times. Hoechst dye Thermo Fisher Scientific was applied for nuclei staining. Data were normalized to the infected control, and the IC 50 value was calculated by GraphPad Prism 7.
In brief, Vero cells were seeded into a six-well plate 24 h prior to infection. A serial dilution of the virus-containing supernatants was prepared for inoculation into the Vero cell monolayer. Thereafter, plaque phenotypes were visualized by staining with 0.
Plaque numbers were counted as plaque-forming units per milliliter and reported as the percentage of plaque reduction in comparison to supernatant obtained from the infected Calu-3 cells without any treatment. Cell viability was examined by an MTT colorimetric assay. In brief, the medium was replaced with MTT [3- 4,5-dimethylthiazolyl -2,5-diphenyltetrazolium bromide] Sigma-Aldrich at a final concentration of 0. Absorbance was measured at a wavelength of nm by an EnVision multilabel reader PerkinElmer.
Data were normalized to the solvent control, and then CC 50 values were calculated using GraphPad Prism 7. We thank the Department of Disease Control, Ministry of Public Health Thailand, for providing the clinical specimens for viral isolation. This research project is supported by Mahidol University, Thailand. All authors analyzed and interpreted the data. All authors read and approved the final version of the manuscript. National Center for Biotechnology Information , U. J Nat Prod. Published online Apr Author information Article notes Copyright and License information Disclaimer.
Corresponding author. Email: moc. Email: ude. Received Dec 8. This article has been cited by other articles in PMC. Open in a separate window. Figure 1. Figure 2. Dose—Response Relationship of A. Figure 3. Table 1 IC 50 Values of A. Cytotoxicity Profiles of A. Figure 4. Figure 5. Potential Clinical Applications of A. Acknowledgments We thank the Department of Disease Control, Ministry of Public Health Thailand, for providing the clinical specimens for viral isolation.
Author Contributions S. Author Contributions K. Notes The authors declare no competing financial interest. References Huang J. Severe acute respiratory syndrome coronavirus as an agent of emerging and reemerging infection. A familial cluster of pneumonia associated with the novel coronavirus indicating person-to-person transmission: a study of a family cluster. Lancet , , — Estimates of the severity of coronavirus disease a model-based analysis.
Lancet Infect. Clinical Characteristics of Coronavirus Disease in China. Clinical features of patients infected with novel coronavirus in Wuhan, China. Agents Chemother.
Nature , , — Antiviral Res. JAMA , 11 , — Remdesivir for the Treatment of Covid - Final Report. Covid Remdesivir has little or no impact on survival, WHO trial shows. Natural Products as Sources of New Drugs from to Plants , 9 9 , Virus Res. Changes in the contents of four active diterpenoids at different growth stages in Andrographis paniculata Burm.
Nees Chuanxinlian. Andrographis paniculata Burm. World J. Thioacetamide induced toxicity in albino rat as manifested by the significant increase in total bilirubin, direct bilirubin, SGPT, SGOT and creatinine while significant decrease in liver protein and kidney protein. These results prove the potential hepatoprotective and renoprotective activity of A.
Keywords: A. Introduction shown to possess hepatoprotective property by improving antioxidant status [15]. Liver cirrhosis is the damage of cells and their gradual replacement with scar tissue that damages blood flow A. It is an herbaceous liver function [1]. Modern pharmacological of metabolic and physiologic homeostasis of the body [2]. It studies have demonstrated its cardioprotective, is involved in all biochemical pathways to growth, fight antithrombolytic, immunostimulant, antihyperglycemic, against disease and nutrient supply.
The carbohydrate, antimalarial and anti-inflammatory properties and so on [16]. Thus In the present study the medicinal plant A. The kidney is an essential organ that plays a dominant role in 2. Material homeostasis by excreting the metabolic waste products and keeps necessary products depending on the needs of the body 2. Kidney damage by various toxicants causes morphological with tubular or interstitial changes to Healthy adult Wistar rats — gm were procured from nephropathy [5].
Hindustan Antibiotic Ltd, Pune and were acclimatized in laboratory condition for two weeks. TAA acts as a sulfur donor and used for various industrial and pharmaceutical application. It is also a 3. Methods carcinogenic [6] [7] [8] [9]. Thioacetamide induces cirrhosis pathological features of which are similar in many aspects to 3.
It is one of numerous agents that cause structural and functional changes TAA Sigma Aldrich, Switzerland was prepared freshly by not only in liver but also in other tissues such as kidney, dissolving in sterile distilled water and stirred until all thymus, spleen and lungs [10].
The long times history of using herbs in 3. In recent decades herbal medicine has received great attention because low costs and Fresh A. The plant Volume 5 Issue 4, April www. Salunkhe, Department of rat was A voucher and in a treated group was The total bilirubin in control, induced and treated rat was 0.
The direct 3. Preparation of extracts bilirubin was 0. The conical flask shake severally covered overnight and stored at room Table 1: Effect of aqueous extract of A. The filtrate was evaporated at C Control Induced Treated upto complete dryness and forms chocolate coloured powder.
Group I: Control group of 3 to 5 months rat with either sex receiving intraperitoneal injection of distilled water for eight week. Group II: Induced group were given i. Figure 1: Variations in conc. Blood sample different group was directly collected from left ventricle and were allowed to clot at room temperature. After clotting the sample was centrifused and serum is obtained on top of tube. Serum was collected for further experiment.
Result Figure 2: Change in conc. On the other hand the increase in these parameters was prevented by treatment of animals with A. Increased level of SGOT was noticed in thioacetamide induction group In treated group SGOT level was found decreased Thioacetamide is a hepatotoxic and produce hepatic necrosis. On metabolism thioacetamide produce free radicals resulting in oxidative stress and induces apoptasis of hepatocytes [24].
On long term taken thioacetamide causes cirrhosis in rats. Thioacetamide induces centrolobular hepatic necrosis, hepatocellular carcinoma, liver cirrhosis and injury to the terminal portion of the proximal renal tubule [25].
These observations are similar to those of [26] [27] [28] [29] who have used thioacetamide to induce liver cirrhosis.
In chronic condition TAA intoxication, Protein in liver and kidney. Bilirubin is whereas in induced group it is increased to Creatinine, a non-protein waste product is freely filtered by the kidney. The serum creatinine concentration is the most commonly used parameter of the kidney function. The level of creatinine in the blood rises if kidney is not function properly.
Current research has focused on alternative approaches to synthetic pharmacology, which rely on natural products. It target on medicinal plants which shows curative effect on diseases of the liver or other organ [33].
Whole parts of A. The creatinine level which was Figure 4: Change in conc. The total protein conc. The treated group shows increased protein treated by aqueous extract of A. Discussion apart from other beneficial features. It contains andrographolides, alkaloids, flavonoids, phenolic compounds Liver shows many important functions one of these is a steroids and saponins.
Some industrial and environmental toxicants and other drugs can cause renal damage due to activation of highly reactive free radicals Volume 5 Issue 4, April www. Conclusion Bioss. It also found that total proteins in liver and kidney are [14] Azadbakht, M. Shabankhani B. Effect of essential oils of Artemisia Aucheri Bioss.
Acknowledgement L. Iran J Pharm Res, 2: The author is grateful to Principal and Head, P. College, Karad, for providing the B. Hepatoprotective and antioxidant properties necessary facilities during present work.
J Pharmacy. Res;3 6 References [16] Ojha, S. Liver Tissue Myocardium of Rats. Global Journal of Pharmacology Engineering. Biomaterial for Tissue Engineering. Hepatoprotective effects of orthosiphon [2] Verma, N. Hepatoprotective stamineus extract on thioacetamide-induced liver activity of leaves of Zanthoxylum aramatum DC in cirrhosis in rats. Indian journal of Alternative Medicine.
Abubakar, M. Shehu, R.
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