Ethyl palmitate, an anti-chikungunya virus principle from Sauropus androgynus, a medicinal plant used to alleviate fever in ethnomedicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Sauropus androgynus is a medicinal shrub used for the treatment of fever in ethnomedical traditions in various Southeast Asian countries. AIM OF THE STUDY: This study was aimed to identify antiviral principles from S. androgynus against Chikungunya virus (CHIKV), a major mosquito-borne pathogen that re-emerged in the last decade, and to unravel their mechanism of action. MATERIALS AND METHODS: Hydroalcoholic extract of S. androgynus leaves was screened for anti-CHIKV activity using cytopathic effect (CPE) reduction assay. The extract was subjected to activity guided isolation and the resultant pure molecule was characterized by GC-MS, Co-GC and Co-HPTLC. The isolated molecule was further evaluated for its effect by plaque reduction assay, Western blot and immunofluorescence assays. In silico docking with CHIKV envelope proteins and molecular dynamics simulation (MD) analyses were used to elucidate its possible mechanism of action. RESULTS: S. androgynus hydroalcoholic extract showed promising anti-CHIKV activity and its active component, obtained by activity guided isolation, was identified as ethyl palmitate (EP), a fatty acid ester. At 1 µg/mL, EP led to 100% inhibition of CPE and a significant 3 log10 reduction in CHIKV replication in Vero cells at 48 h post-infection. EP was highly potent with an EC50 of 0.0019 µg/mL (0.0068 µM) and a very high selectivity index. EP treatment significantly reduced viral protein expression, and time of addition studies revealed that it acts at the stage of viral entry. A strong binding to the viral envelope protein E1 homotrimer during entry, thus preventing viral fusion, was identified as a possible mechanism by which EP imparts its antiviral effect. CONCLUSIONS: S. androgynus contains EP as a potent antiviral principle against CHIKV. This justifies the use of the plant against febrile infections, possibly caused by viruses, in various ethnomedical systems. Our results also prompt more studies on fatty acids and their derivatives against viral diseases.

Bioassay guided fractionation of Cyclea peltata using in vitro RAW 264.7 cell culture, antioxidant assays and isolation of bioactive compound tetrandrine.

BACKGROUND: Cyclea peltata is one of the herbs mentioned in ancient scriptures of Ayurveda and is used in different types of Ayurvedic gritham preparations. Moreover, in traditional/tribal medicine C. peltata is used as digestive, anti-inflammatory, diuretic and to treat jaundice, digestive disorders, etc. OBJECTIVE: Activity guided fractionation of C. peltata and in correlation with the levels of bioactive compound tetrandrine. MATERIALS AND METHODS: Preliminary phytochemical screening, estimation of total alkaloid content, preparation of different extracts of C. peltata (crude extract CP, hexane extract HCP, chloroform extract CCP, methanol extract MCP, alkaloid fraction ACP). In vitro anti-inflammatory studies using RAW 264.7 cells and in vitro antioxidant assays of the different extracts of C. peltata. HPTLC estimation of tetrandrine (TET) was carried out using solvent system toluene: ethyl acetate: diethylamine (7.2: 2: 0.8) and isolation of TET from ACP. RESULTS: Preliminary phytochemical studies of C. peltata showed the presence of alkaloid content in all extracts. Whereas, saponins, steroids and terpenoids were detected in CP and CCP. ACP and TET showed significant in vitro anti-inflammatory and antioxidant activity when compared to other extracts. ACP and TET (100 µg/ml) treatment significantly inhibited the mRNA expression of iNOS, COX-2, TNF-α in LPS treated RAW 264.7 cells. HPTLC estimation of bioactive compound tetrandrine was highest in ACP-228.4 µg/mg followed by CP-29.62 µg/mg, CCP-23.46 µg/mg, MCP-18.82 µg/mg and HCP-1.25 µg/mg. TET has been isolated from ACP. CONCLUSION: The results of the present in vitro assays revealed that the alkaloid fraction (ACP) is the most active fraction when compared to other extracts and has a positive correlation with the levels of bioactive compound tetrandrine.

Protective effect of ethyl acetate fraction of Drynaria quercifolia against CCl4 induced rat liver fibrosis via Nrf2/ARE and NFκB signalling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Drynaria quercifolia rhizome is traditionally used as hepatoprotective drug especially in chronic jaundice. AIM OF THE STUDY: The present study was undertaken to scientifically evaluate the efficacy of D. quercifolia rhizome against liver fibrosis. MATERIALS AND METHODS: D. quercifolia rhizome crude extract (DQ) and its fractions of hexane (HDQ), ethyl acetate (EDQ), butanol (BDQ) were evaluated in vitro using primary hepatocytes and RAW 264.7 cells. In vivo anti-liver fibrotic activity of EDQ was assessed using CCl4 induced liver fibrosis in Wistar rats and serum biochemical parameters (AST, ALT, ALP, SB, cholesterol), MDA, PT, INR, GSH, SOD, CAT, liver glycogen, serum albumin levels were monitored. qRT-PCR analysis of TNF-α, COX-2, iNOS were performed. ELISA method was used to estimate TNF-α, COX-1 & 2. Histopathological studies like H & E, Masson's trichrome, immunohistochemistry staining for α-SMA, TIMP-1, Nrf2 were conducted. LC-Q-TOF-MS analysis of EDQ was conducted. RESULTS: In vitro activity guided fractionation of D. quercifolia revealed EDQ as active fraction when compared to other extracts. EDQ treatment significantly inhibited the expression of α-SMA, TIMP-1, COX-2, TNF-α, iNOS and increased the levels of Nrf2 in rat liver fibrosis. LC-Q-TOF-MS analysis of EDQ confirmed the presence of naringin and naringenin. CONCLUSION: The anti-liver fibrotic activity of EDQ is via inhibition of NFκB signalling pathway, antioxidant response through Nrf2 activation and further inhibition of HSC activation.

An Exploration of Phytochemicals from Simaroubaceae

Natural products such as plants, animals and minerals have been the basis of treatment of human diseases. Herbal remedies have been used for the treatment of many ailments. Many compounds have been derived from the plant species mentioned in the ancient texts of Indian system of medicine for the treatment of a number of ailments. The R and D thrust in the pharmaceutical sector is focused on development of new drugs, innovative/indigenous processes for known drugs and development of plant based drugs through investigation of leads from the traditional systems of medicine. The family Simaroubaceae is grouped in the order Rutales, is known to have a diverse range of secondary metabolites. Plants from this family are used as medicine to cure cancer and many other diseases. Isolation of diverse chemical compounds from Simaroubaceae on its stem bark, root bark and leaves have been reported. In this review, we are analysing with the chemical constituents of family Simaroubaceae.

Hepatoprotective activity of Oxalis corniculata L. ethanolic extract against paracetamol induced hepatotoxicity in Wistar rats and its in vitro antioxidant effects.

Oxalis corniculata is well known for its medicinal properties like anti-inflammatory, digestive, diuretic, antibacterial, antiseptic etc. The present study focuses on the ability of O. corniculata to alleviate liver damage caused by over dose of paracetamol. Antioxidant activity of O. corniculata was evaluated using the free radical scavenging activity of 1, 1-diphenyl-2- picrylhydrazyl radicals, total anti oxidant capacity by phosphomolybdenum method and total phenolic content was also evaluated. The ethanolic extract of whole plant of O. corniculata (OC, 500 microg/mL, po) significantly reduced 1, 1-diphenyl-2- picrylhydrazyl radicals. This dose also caused significant reduction (62.67%) in malondialdehyde levels of murine hepatic tissues. The antioxidant capacity of OC was comparable to that of standard ascorbic acid and showed 53.5 microg of phenol/mg OC. Rats pre-treated with OC for 4 days showed significant reduction in the serum enzymes such as glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, alkaline phosphatase, serum bilirubin and showed almost normal histological liver architecture of the treated groups compared to paracetamol induced hepatic damage group, indicating its hepatoprotective and antioxidant potential.

Antioedematous and Analgesic Properties of Fertile Fronds of Drynaria quercifolia.

Inflammation is a complex biological response of tissue cells to harmful stimuli including trauma, tissue necrosis, and infections which plays a key role in the pathophysiology of many deadly diseases. In ethnomedicine Drynaria quercifolia fronds are used to treat inflammation as poultice on swellings and as antibacterial, hepatoprotective, and antipyretic agent. Herein, we have evaluated the antioedematous, antiproliferative, and analgesic properties of the ethanolic extract of fertile fronds of D. quercifolia (FF) by standard procedures. Oral administration of FF produced significant inhibition of carrageenan and histamine induced paw oedema in Wistar rats. FF significantly reduced both wet weight and dry weight of granuloma tissue which shows the inhibitory effect on exudative and proliferative phases of inflammation. FF significantly attenuated acute and delayed phases of formalin induced pain, acetic acid-induced writhing, capsaicin-induced nociception, and hot plate test in mice. Phytochemical analysis revealed the presence of coumarins, flavonoids, glycosides, phenolics, saponins, steroids, tannins, and terpenoids. Total phenolic content was 186 mg/g equivalent of gallic acid. The HPLC estimation showed flavanone glycoside naringin (1.2%) and its aglycone naringenin (0.02%). The presence of potent anti-inflammatory and analgesic principles in FF and their synergistic action may be the reason for the proposed therapeutic effects.

Anti-inflammatory and analgesic properties of Drynaria quercifolia (L.) J. Smith.

ETHNOPHARMACOLOGICAL RELEVANCE: Drynaria quercifolia (L.) J. Smith (Polypodiaceae), has been widely used by ethnic groups of India to treat inflammation, rheumatism, headache, bone fracture, jaundice, etc. AIM OF THE STUDY: To evaluate the anti-inflammatory and analgesic properties of the ethanolic extract of rhizome of Drynaria quercifolia (DQ) and its phytochemical profile. MATERIALS AND METHODS: DQ was used to evaluate the anti-inflammatory and analgesic effects using carrageenan-induced paw oedema/cotton pellet-induced granuloma in Wistar rats and acetic acid-induced writhing/formalin-induced paw licking test in Swiss albino mice respectively. RESULTS: Oral administration of DQ produced significant inhibition of carrageenan-induced paw oedema and granuloma formation in rats, almost comparable to that caused by indomethacin. DQ significantly attenuated acute and delayed phases of formalin-induced pain and acetic acid-induced writhing episodes in mice. The analgesia was comparable to that produced by sodium salicylate and aspirin respectively. Phytochemical analysis gave positive tests for catechin, coumarins, flavonoids, phenolics, saponin, steroids, tannins, and triterpenes. The total phenolics in DQ was 244 mg/g and naringin content was 0.048%. CONCLUSION: The results suggest the presence of potent anti-inflammatory and analgesic principles in DQ that justifies its use for alleviating painful inflammatory conditions.

Anti-allergic, anti-inflammatory and anti-lipidperoxidant effects of Cassia occidentalis Linn.

Cassia occidentalis Linn. mast cell degranulation at a dose of 250 mg/kg, showed dose dependent stabilizing activity towards human RBC, with is widely used in traditional medicine of India to treat a number of clinical conditions including allergy and inflammatory manifestations. In the present study anti-allergic, anti-inflammatory and anti-oxidant properties of C. occidentalis whole plant ethanolic extract (CO) was investigated. Effects of CO on rat mast cell degranulation inhibition and human red blood cell (HRBC) membrane stabilization were studied in vitro following standard methods. The anti lipidperoxidant effects of CO were also studied in vitro. Effect of CO on carrageenan-induced mouse paw oedema inhibition was also assessed. CO significantly decreased maximum protection of 80.8% at 15 microg/ml. The extract also caused significant reduction in malondialdehyde (MDA) levels of murine hepatic microsomes at 100 microg/ml (56%) and significantly reduced carrageenan induced inflammation in mice at a dose of 250 mg/kg. Results of the present study indicated that CO inhibited mast cell degranulation, stabilized HRBC membrane thereby alleviating immediate hypersensitivity besides showing anti oxidant activity.

Hepatoprotective effect of three herbal extracts on aflatoxin B1-intoxicated rat liver.

INTRODUCTION: Roots of Ixora coccinea (Rubiaceae), and Rhinacanthus nasuta (Acanthaceae) and whole plants of Spilanthes ciliata (Asteraceae) are extensively used by tribal communities in South India to treat liver diseases. However, the veracity of these tribal claims has not been investigated scientifically using the liver toxin, aflatoxin. This study reports on the protective effects of these three herbal ethanolic extracts on the aflatoxin B1 (AFB1)-intoxicated livers of albino male Wistar rats. METHODS: Biochemical parameters, including serum hepatic enzymes (glutamate oxaloacetate transaminase, glutamate pyruvate transaminase and alkaline phosphatase), were studied. Hepatic tissues were processed for assay of reduced glutathione (GSH) and histological alterations. RESULTS: Pre-treatment of the rats with oral administration of the plant ethanolic extracts, Ixora coccinea (IC), Rhinacanthus nasuta (RN), Spilanthes ciliata (SC), prior to AFB1 was found to provide significant protection against toxin-induced liver damage, determined 72 hours after the AFB1 challenge (1.5 mg/kg, intraperitoneally) as evidenced by a significant lowering of the activity of the serum enzymes and enhanced hepatic reduced GSH status. Pathological examination of the liver tissues supported the biochemical findings. The three plant extracts, IC, RN and SC, showed significant antilipid peroxidant effects in vitro. CONCLUSION: It was concluded that the hepatoprotective effects of the three plant extracts observed in this study might result from their potent antioxidative properties.

Hepatoprotective studies on Sida acuta Burm. f.

ETHNOPHARMACOLOGICAL RELEVANCE: Sida acuta Burm. f. (Malvaceae) is used in Indian traditional medicine to treat liver disorders and is useful in treating nervous and urinary diseases and also disorders of the blood and bile. AIM OF THE STUDY: Evaluation of the hepatoprotective properties of the methanolic extract of the root of Sida acuta (SA) and the phytochemical analysis of SA. MATERIALS AND METHODS: The model of paracetamol-induced hepatotoxicity in Wistar rats, liver histopathological observations, hexobarbitone-induced narcosis and in vitro anti-lipid peroxidation studies were employed to assess the hepatoprotective efficacy of SA. Phytochemical assay of SA was conducted following standard protocols. RESULTS: Significant hepatoprotective effects were obtained against liver damage induced by paracetamol overdose as evident from decreased serum levels of glutamate pyruvate transaminase, glutamate oxaloacetate transaminase, alkaline phosphatase and bilirubin in the SA treated groups (50, 100, 200mg/kg) compared to the intoxicated controls. The hepatoprotective effect was further verified by histopathology of the liver. Pretreatment with Sida acuta extract significantly shortened the duration of hexobarbitone-induced narcosis in mice indicating its hepatoprotective potential. Phytochemical studies confirmed the presence of the phenolic compound, ferulic acid in the root of Sida acuta, which accounts for the significant hepatoprotective effects observed in the present study. CONCLUSION: The present study thus provides a scientific rationale for the traditional use of this plant in the management of liver disorders.

Gastric antisecretory and antiulcer activities of Cyclea peltata (Lam.) Hook. f. & Thoms. in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Cyclea peltata (Lam.) Hook. f. & Thoms. (Menispermaceae), locally called 'Padathaali/Padakizhangu' is used in Ayurvedic medicine to treat peptic ulcer. AIM OF THE STUDY: To evaluate the gastric antisecretory and antiulcer activity of Cyclea peltata. MATERIALS AND METHODS: The ethanolic extract of Cyclea peltata root was used to evaluate its gastric antisecretory and antiulcer effect in the pylorus-ligated rat model and gastric lesions induced by ethanol or ethanol and indomethacin respectively in rats. The levels of gastric wall mucus, non-protein sulfhydryl groups (NP-SH), malondialdehyde, protein and catalase activity in the stomach samples of Cyclea peltata treated and control groups of rats were also quantified. RESULTS: The ethanolic extract of Cyclea peltata roots showed significant antisecretory activity as evidenced by decreased pepsin secretion, gastric juice volume and acid output in pylorus-ligated rats. Pretreatment with Cyclea peltata extract provided significant protection against the peptic ulceration caused by ethanol administered individually, or in combination with indomethacin. Our studies also revealed that pretreatment with Cyclea peltata significantly increased the gastric protein and catalase concentration of ethanol treated rats. Further, it showed significant gastroprotective effects on the stomach wall of ethanol or ethanol and indomethacin treated rats by decreasing malondialdehyde level, increasing the gastric wall mucus and non-protein sulfhydryl groups. CONCLUSION: The present findings demonstrate that Cyclea peltata ethanolic extract has potent antisecretory and antiulcer effects and justify the traditional/ethnic usage of this herb to treat peptic ulcers and consequent stomach ache.

Hepatoprotective effect of Hibiscus hispidissimus Griffith, ethanolic extract in paracetamol and CCl4 induced hepatotoxicity in Wistar rats.

Hibiscus hispidissimus Griff. is used in tribal medicine of Kerala, the southern most state of India, to treat liver diseases. In the present study, the effect of the ethanolic extract of Hibiscus hispidissimus whole plant on paracetamol (PCM)-induced and carbon tetrachloride (CCl4)-induced liver damage in healthy Wistar albino rats was studied. The results showed that significant hepatoprotective effects were obtained against liver damage induced by PCM and CCl4 as evidenced by decreased levels of serum enzymes, glutamate oxaloacetate transaminase (SGOT), glutamate pyruvate transaminase (SGPT), serum alkaline phosphatase (SAKP), serum bilirubin (SB) and an almost normal histological architecture of the liver of the treated groups compared to the toxin controls. The extract also showed significant antilipid peroxidant effects in vitro, besides exhibiting significant activity in quenching 1, 1-diphenyl-2-picryl hydrazyl (DPPH) radical, indicating its potent antioxidant effects.

Hepatoprotective effects of Pittosporum neelgherrense Wight&Arn., a popular Indian ethnomedicine.

The stem bark of Pittosporum neelgherrense Wight&Arn. is used by the Kani and Malapandaram tribes of Kerala as an effective antidote to snake bite and for the treatment of various hepatic disorders. In the present study, the effect of the methanolic extract of the stem bark of Pittosporum neelgherrense was studied against carbon tetrachloride (CCl(4))-, d-galactosamine (D-GalN)- and acetaminophen (APAP)-induced acute hepatotoxicity in Wistar rats. Significant hepatoprotective effects were obtained against liver damage induced by all the three liver toxins, as evident from decreased levels of serum enzymes, glutamate oxaloacetate transaminase (SGOT), glutamate pyruvate transaminase (SGPT) and an almost normal architecture of the liver in the treated groups, compared to the toxin controls. Thus the present study provides a scientific rationale for the traditional use of this plant in the management of liver diseases.

Evaluation of hepatoprotective effects of Helminthostachys zeylanica (L.) Hook against carbon tetrachloride-induced liver damage in Wistar rats.

The rhizomes of Helminthostachys zeylanica (L.) are used by the Kattunaikan tribe of Kerala, for the treatment of various hepatic disorders. In the present study, the effect of the methanolic extract of Helminthostachys zeylanica rhizomes on carbon tetrachloride (CCl4)-induced liver damage in Wistar rats was studied. The results showed that significant hepatoprotective effect was obtained against CCl4-induced liver damage, by oral administration of Helminthostachys zeylanica methanolic extract as evident from decreased levels of serum enzymes and an almost normal architecture of the liver, in the treated groups, compared to the controls. The extract was effective in increasing the choleretic activity of anaesthetised normal rats. It also shortened hexobarbitone-induced sleeping time in mice, which was increased by CCl4 treatment, besides showing significant antilipid peroxidant effect in vitro. Thus, the present study provides a scientific rationale for the traditional use of this plant in the management of liver diseases.

Modulatory effects of lxora coccinea flower on Cyclophosphamidetoxicity in tumour bearing mice.

The active fraction (AF) from lxora coccinea flowers prevented the decrease in haemoglobin levels and leucocyte counts of Dalton's lymphoma tumour bearing mice, treated with cyclophosphamide (CYP). It also significantly increased the life span of tumour bearing mice, treated with cyclophosphamide. Serum glutamate pyruvate transaminase (SGPT) and serum alkaline phosphatase (SAKP) levels of tumour bearing mice treated with CYP were decreased significantly by combination therapy with I.coccinea AF. indicating protection against hepatic toxicity.