In vitro Evaluation of Anti-Cancer Potential of Different Solvent Extracts Derived from Clerodendrum Infortunatum Linn against Cervical Cancer.

BACKGROUND: Cervical cancer is a prevalent and deadly malignancy in females, with chemotherapy often proving ineffective due to significant side effects and the development of chemo-resistance. This study investigates the medicinal potential of Clerodendrum infortunatum linn. , a genus with approximately 500 species in the Lamiaceae family. Limited research exists on the species of Clerodendrum infortunatum and its various solvent extracts. OBJECTIVE: The study aims to assess the anti-cancer properties of different solvent extracts from this plant on human cervical cancer cells. METHODS: The study examines the plant's phytochemical components and their potential to inhibit cancer growth. Aerial parts of the plant were extracted using the Soxhlet method, and the presence of Rutin, Quercetin, and Gallic Acid in specific solvent extracts was validated through High-Performance Thin Layer Chromatography (HPTLC). In vitro assays, including MTT, Apoptosis, Cell Cycle analysis, Intracellular Reactive Oxygen Species assessment, and Gene expression PCR, were conducted to investigate the plant's anti-cancer properties further. RESULTS: The outcomes of the phytochemical assessment indicated that Rutin was predominantly present in the water extract, with quercetin being more concentrated in the decoction, and the hydro-alcoholic extract showing elevated levels of gallic acid. Notably, the decoction extract demonstrated the highest cytotoxic activity, primarily through early apoptosis and arrests in the S-phase and G2M phases. Clerodendrum infortunatum exhibited a reduction in Intracellular Reactive Oxygen Species. The gene expression analysis disclosed an impact on the BCL-2 gene. CONCLUSION: Notably, Clerodendrum infortunatum exhibited the ability to initiate early apoptosis, halt the cell cycle at the S and G2M phases, and diminish levels of reactive oxygen species significantly. The gene expression analysis revealed an influence on the BCL-2 gene. To sum up, this research underscores the encouraging cytotoxic and antioxidant attributes of Clerodendrum infortunatum, implying its potential for cervical cancer treatment.

Polyphenol Mediated Suppression of Hepatocellular Carcinoma (HepG2) Cell Proliferation by Clerodendrum infortunatum L. Root.

OBJECTIVE: Clerodendrum infortunatum L. has long been used in traditional medicine in Sri Lanka for tumours, cancer, and certain skin diseases. The present study aimed to assess the anticancer properties of the aqueous extract of C. infortunatum L. root (AECIR) through the activation of the apoptotic pathway on hepatocellular carcinoma (HepG2) and thus give it a scientific validation. Further, the contribution of polyphenols in antioxidant activity and cell cytotoxicity was investigated. METHODS: Powdered plant material was boiled with water (100°C) to obtained AECIR.  The DPPH assay was used to determine the antioxidant potential. The activity of AECIR on HepG2 and normal rat fibroblast (CC1) cell growth was determined using MTT assay. The morphological changes related to apoptotic pathway was examined by Ethidium Bromide/Acridine Orange (EB/AO), Rhodamine 123 (Rh123) and DNA fragmentation assay. RESULTS: The AECIR demonstrated antioxidant potential with an EC50 of 350.2 ± 1.5 ug/mL for DPPH assay. The HO•, H2O2 and •NO free radical scavenging activity was observed with EC50 of 19.7 ± 2.3, 11.7 ± 0.1 and 273.1 ± 0.9 ug/mL, respectively. The antiproliferative effect of AECIR on HepG2 cells was observed in a time and dose dependent manner with an EC50 of 239.1 ± 1.3 µg/mL while CC1 cells showed a nontoxic effect with an EC50 1062.7 ± 3.4 µg/mL after 24hrs treatment. A significant decrease in antioxidant activity (p<0.001) and 90% HepG2 cell viability was observed with polyphenol removed AECIR compared to the polyphenol present AECIR. The EB/AO uptake, depletion of mitochondrial transmembrane potential, and DNA fragmentation assay results revealed that the apoptosis was induced by AECIR. CONCLUSION: The obtained result of the present study demonstrates that the antioxidant potential and antiproliferative activity of AECIR is attributed to the presence of polyphenols. Furthermore, the findings provide the scientific base for anti-cancer potential of AECIR.

Clerodendrum chinense Stem Extract and Nanoparticles: Effects on Proliferation, Colony Formation, Apoptosis Induction, Cell Cycle Arrest, and Mitochondrial Membrane Potential in Human Breast Adenocarcinoma Breast Cancer Cells.

Breast cancer stands out as the most widespread form of cancer globally. In this study, the anticancer activities of Clerodendrum chinense (C. chinense) stem ethanolic extract were investigated. High-performance liquid chromatography (HPLC) analysis identified verbascoside and isoverbascoside as the major bioactive compounds in the C. chinense stem extract. Successfully developed nanoparticles exhibited favorable hydrodynamic diameter, polydispersity index, and surface charge, thus ensuring stability after four months of storage. The total phenolic content and total flavonoid contents in the nanoparticles were reported as 88.62% and 95.26%, respectively. The C. chinense stem extract demonstrated a dose-dependent inhibitory effect on MCF-7, HeLa, A549, and SKOV-3 cancer cell lines, with IC50 values of 109.2, 155.6, 206.9, and 423 µg/mL, respectively. C. chinense extract and NPs exhibited dose-dependent cytotoxicity and the highest selectivity index values against MCF-7 cells. A dose-dependent reduction in the colony formation of MCF-7 cells was observed following treatment with the extract and nanoparticles. The extract induced cytotoxicity in MCF-7 cells through apoptosis and necrosis. C. chinense stem extract and nanoparticles decreased mitochondrial membrane potential (MMP) and induced G0/G1 phase arrest in MCF-7 cells. In conclusion, use of C. chinense stem extract and nanoparticles may serve as a potential therapeutic approach for breast cancer, thus warranting further exploration.

Clerodendrum viscosum leaves attenuate lead-induced neurotoxicity through upregulation of BDNF-Akt-Nrf2 pathway in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Clerodendrum viscosum is an important medicinal plant in Ayurveda in Bangladesh and its leaves are used as a remedy for various diseases such as anti-inflammatory, antibacterial, hyperglycemic, hepatoprotective effects. AIM OF THE STUDY: The present study aimed to evaluate the protective effect of aqueous extract of C. viscosum leaves against Pb-induced neurobehavioral and biochemical changes in mice. MATERIALS AND METHODS: Swiss albino mice were divided as a) control, b) lead treated (Pb) and c) C. viscosum leaves (Cle) d) Pb plus Cle groups. Pb-acetate (10 mg/kg body weight) was given to Pb and Pb + Cle groups mice, and water extract of leaves (50 mg/kg body weight) was provided as supplementation to Cle and Pb + Cle groups mice for 30 days. Elevated plus maze and Morris water maze tests were used for evaluating anxiety, spatial memory and learning, respectively. Status of cholinesterase, SOD, GSH enzyme activity and neurotoxicity markers such BDNF and Nrf2 levels were analyzed in the brain tissue of experimental mice. RESULTS: Poorer learning, inferior spatial memory, and increased anxiety-like behavior in Pb-exposure mice were noted when compared to control mice in Morris water maze and elevated plus maze test, respectively. In addition, expression of BDNF and Nrf2, cholinesterase activity along with antioxidant activity were significantly reduced compared to control group (p < 0.01). Interestingly, C. viscosum leaves' aqueous extract supplementation in Pb-exposed mice provide a significant improved neurochemical and antioxidant properties through the augmentation of activity of cholinergic enzymes, and upregulation of BDNF and Nrf2 levels in the brain tissue compared to Pb-exposed mice. CONCLUSIONS: This study suggested that C. viscosum leaves restore the cognitive dysfunction and reduce anxiety-like behavior through upregulation of BDNF mediated Akt-Nrf2 pathway in Pb-exposure mice.

Hesperetin from Root Extract of Clerodendrum petasites S. Moore Inhibits SARS-CoV-2 Spike Protein S1 Subunit-Induced NLRP3 Inflammasome in A549 Lung Cells via Modulation of the Akt/MAPK/AP-1 Pathway.

Inhibition of inflammatory responses from the spike glycoprotein of SARS-CoV-2 (Spike) by targeting NLRP3 inflammasome has recently been developed as an alternative form of supportive therapy besides the traditional anti-viral approaches. Clerodendrum petasites S. Moore (C. petasites) is a Thai traditional medicinal plant possessing antipyretic and anti-inflammatory activities. In this study, C. petasites ethanolic root extract (CpEE) underwent solvent-partitioned extraction to obtain the ethyl acetate fraction of C. petasites (CpEA). Subsequently, C. petasites extracts were determined for the flavonoid contents and anti-inflammatory properties against spike induction in the A549 lung cells. According to the HPLC results, CpEA significantly contained higher amounts of hesperidin and hesperetin flavonoids than CpEE (p < 0.05). A549 cells were then pre-treated with either C. petasites extracts or its active flavonoids and were primed with 100 ng/mL of spike S1 subunit (Spike S1) and determined for the anti-inflammatory properties. The results indicate that CpEA (compared with CpEE) and hesperetin (compared with hesperidin) exhibited greater anti-inflammatory properties upon Spike S1 induction through a significant reduction in IL-6, IL-1ß, and IL-18 cytokine releases in A549 cells culture supernatant (p < 0.05). Additionally, CpEA and hesperetin significantly inhibited the Spike S1-induced inflammatory gene expressions (NLRP3, IL-1ß, and IL-18, p < 0.05). Mechanistically, CpEA and hesperetin attenuated inflammasome machinery protein expressions (NLRP3, ASC, and Caspase-1), as well as inactivated the Akt/MAPK/AP-1 pathway. Overall, our findings could provide scientific-based evidence to support the use of C. petasites and hesperetin in the development of supportive therapies for the prevention of COVID-19-related chronic inflammation.

Cytoprotective effects and antioxidant activities of acteoside and various extracts of Clerodendrum cyrtophyllum Turcz leaves against t-BHP induced oxidative damage.

This study evaluates the antioxidant potential and cytoprotective effects of ethanolic crude extract from Clerodendrum cyrtophyllum leaves (ECE) and five derived fractions (namely, petroleum ether fraction (PEF), dichloromethane fraction (DMF), ethyl acetate fraction (EAF), n-butyl alcohol fraction (BAF) and the remaining fraction (RF)), as well as acteoside (Ac, a major phenolic component in EAF) on oxidative damage caused by tert-butyl hydroperoxide (t-BHP) in HepG2 cells. MTT assay results showed that ECE, EAF, BAF, RF and Ac increased the viability of t-BHP-damaged cells in a dose-dependent manner, while EAF significantly promoted cell viability. EAF, BAF, RF, or Ac reduced the levels of lactate dehydrogenase (LDH) leakage, malondialdehyde (MDA), and reactive oxygen species (ROS). Additionally, glutathione (GSH) levels and the activities of superoxide dismutase (SOD) and catalase (CAT) increased. Western blot analysis further indicated that EAF, BAF, RF, or Ac up-regulated pro-caspase-3 and reduced cleaved caspase-3 during t-BHP-induced oxidative stress. Flow cytometry analysis and fluorescence micrographs showed that Ac could inhibit apoptosis.

Seasonal variation of verbascoside as a principal phenolic compound linked with antioxidant potentials of Clerodendrum glandulosum Lindl. leaves.

This study reports the influence of seasonality on the accumulation of verbascoside as a principal phenolic compound in Clerodendrum glandulosum Lindl. leaves along with possible alteration of antioxidant potentials. Leaves were collected during winter (December 2018), spring (February 2019), summer (May 2019), monsoon (July 2019), autumn (October 2019), and extracted with 95% aqueous methanol by cold maceration. The total phenolic content and antioxidant capacities (DPPH, ABTS and FRAP) were estimated by spectrophotometric technique, and verbascoside content was estimated by HPLC-PDA. Results indicate that the leaves collected during summer and winter both exhibited the highest total phenolic content verbascoside accumulation and antioxidant potentials which are significantly different (p < 0.05) than other seasons. Correlation studies further demonstrated that the total polyphenol and verbascoside contents were directly proportional to the antioxidant potentials. Thus, the study concludes that winter and summer are the best seasons for collecting leaves from this plant to obtain maximum antioxidant potential.

Evaluation of anticancer activity of Clerodendrum viscosum leaves against breast carcinoma.

INTRODUCTION: The use of natural resources as medicines for cancer therapies has been described throughout history in the form of traditional medicines. However, many resources are still unidentified for their potent biological activities. Clerodendrum viscosum is a hill glory bower reported as a remedy against oxidative stress, skin diseases, and intestinal infections. MATERIALS AND METHODS: We have collected the C. viscosum leaves and used for the preparation of 70% methanolic extract (CVLME). Then, CVLME has been confirmed for anticancer properties on various cancer cell lines by evaluating cytotoxicity, cell cycle analysis, induction of ROS and apoptosis, and nuclear fragmentation. Further, the phytochemical analysis of CVLME was evaluated through high-performance liquid chromatography. RESULTS: Cell proliferation assay revealed the selective cytotoxicity of CVLME against breast cancer cell line (MCF-7). The FACS-based cell cycle analysis showed increased subG1 (apoptosis) population dose dependently. Further, the apoptosis-inducing effect of CVLME was confirmed by annexin staining. Flow cytometry and confocal microscopy revealed the selective ROS generation upon CVLME treatment. The confocal-based morphological study also revealed condensed and fragmented nuclear structure in CVLME-treated MCF-7 cells. Phytochemical investigations further indicated the presence of tannic acid, catechin, rutin, and reserpine which might be the reason for the anticancer activity of CVLME. CONCLUSION: The above-combined results revealed the anticancer effect of CVLME, which may be due to the selective induction of ROS in breast carcinoma.

Antiviral iridoid glycosides from Clerodendrum myricoides.

The methanol root extract of Clerodendrum myricoides (Hochst.) Vatke afforded two new (1, 2) and two known (3, 4) iridoid glycosides. The structures of the isolated compounds were established based on NMR, IR, UV and MS data analyses. The crude extract and the isolated constituents were assayed for antiviral activity against the human respiratory syncytial virus (RSV) in human laryngeal epidermoid carcinoma (HEp-2) cells. The crude extract inhibited RSV infectivity at EC50 = 0.21 µg/ml, while it showed cytotoxicity against HEp-2 cells with CC50 = 9 µg/ml. Compound 2 showed 43.2% virus inhibition at 100 µM, while compounds 1 as well as 3 and 4 had only weak antiviral and cytotoxic activities.

Bioactive Abietane-Type Diterpenoid Glycosides from Leaves of Clerodendrum infortunatum (Lamiaceae).

In this study, two previously undescribed diterpenoids, (5R,10S,16R)-11,16,19-trihydroxy-12-O-ß-d-glucopyranosyl-(1→2)-ß-d-glucopyranosyl-17(15→16),18(4→3)-diabeo-3,8,11,13-abietatetraene-7-one (1) and (5R,10S,16R)-11,16-dihydroxy-12-O-ß-d-glucopyranosyl-(1→2)-ß-d-glucopyranosyl-17(15→16),18(4→3)-diabeo-4-carboxy-3,8,11,13-abietatetraene-7-one (2), and one known compound, the C13-nor-isoprenoid glycoside byzantionoside B (3), were isolated from the leaves of Clerodendrum infortunatum L. (Lamiaceae). Structures were established based on spectroscopic and spectrometric data and by comparison with literature data. The three terpenoids, along with five phenylpropanoids: 6'-O-caffeoyl-12-glucopyranosyloxyjasmonic acid (4), jionoside C (5), jionoside D (6), brachynoside (7), and incanoside C (8), previously isolated from the same source, were tested for their in vitro antidiabetic (α-amylase and α-glucosidase), anticancer (Hs578T and MDA-MB-231), and anticholinesterase activities. In an in vitro test against carbohydrate digestion enzymes, compound 6 showed the most potent effect against mammalian α-amylase (IC50 3.4 ± 0.2 µM) compared to the reference standard acarbose (IC50 5.9 ± 0.1 µM). As yeast α-glucosidase inhibitors, compounds 1, 2, 5, and 6 displayed moderate inhibitory activities, ranging from 24.6 to 96.0 µM, compared to acarbose (IC50 665 ± 42 µM). All of the tested compounds demonstrated negligible anticholinesterase effects. In an anticancer test, compounds 3 and 5 exhibited moderate antiproliferative properties with IC50 of 94.7 ± 1.3 and 85.3 ± 2.4 µM, respectively, against Hs578T cell, while the rest of the compounds did not show significant activity (IC50 > 100 µM).

Chemical compositions and repellent activity of Clerodendrum bungei Steud. essential oil against three stored product insects.

BACKGROUND: Several species of Verbenaceae have been widely used in medicine, and some species of Verbenaceae have been observed good insecticidal activity, such as Lantana camara and Vitex negundo. There is no report about repellent activity of Clerodendrum bungei Steud. (C. bungei) against stored product insects. The chemical composition of C. bungei essential oil (EO) were identified, repellent activity of methanol extract, EO of C. bungei and two main components of EO against T. castaneum, L. serricorne and L. bostrychophila were evaluated for the first time. RESULTS: EO of C. bungei was obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry (GC-MS) and GC. A total of 25 components of the C. bungei EO were identified. The principal compounds in the EO were myristicin (75.0%), 2,2,7,7-Tetramethyltricyclo[6.2.1.0(1,6)]undec-4-en-3-one (4.1%) and linalool (3.4%). Results of bioassays indicated that C. bungei EO exerted strong repellent activity against three target insects. As main constituents, myristicin and linalool also had certain repellency. CONCLUSION: This work suggests that the EO of C. bungei has promising potential to develop into botanical repellents for the control of pest damage in warehouses and grain stores.

Scientific validation of the antimicrobial and antiproliferative potential of Clerodendrum serratum (L.) Moon, its phytoconstituents and their biosafety by acute oral toxicity study.

The screening of aqueous extract of Clerodendrum serratum revealed its broad-spectrum antimicrobial potential against Gram positive, Gram negative bacteria, and yeast. Optimizing the extraction strategies, revealed 15% concentration of aqueous extract prepared at 40 °C by extracting for 40 min, as optimum parameters and its statistical optimization by Box-Behnken design led to 1.16-1.35 folds enhancement in activity. Organic solvent extraction further improved the activity where methanol proved to be the best organic extractant which was effective against all the 13 pathogens tested with inhibition zone ranging from 14 to 32 mm. Minimum Inhibitory Concentration (MIC) study endorsed the methanolic extract to be the best organic extractant, as it showed the lowest MIC (0.5-10 mg/ml) in comparison to aqueous extract (1-10 mg/ml) as well as Partially Purified Phytoconstituents i.e., flavonoids (1-5 mg/ml), diterpenes (5-10 mg/ml) and cardiac glycosides (5-10 mg/ml). All these were found to be biosafe in both In-vitro (Ames and MTT assay) and In-vivo toxicity studies. Acute oral toxicity testing of flavonoids (2000 mg/ml) on Wistar rats did not reveal any significant change in relative organ weight, biochemical, hematological parameters and organs' architecture in comparison to control. Antiproliferative potential of flavonoids against human cancerous cell lines i.e., HeLa, HCT-15, and U87-MG, further increase the importance of this plant as a promising candidate for drug development. The overall study justified the medicinal importance of this plant.

A natural flavonoid, apigenin isolated from Clerodendrum viscosum leaves, induces G2/M phase cell cycle arrest and apoptosis in MCF-7 cells through the regulation of p53 and caspase-cascade pathway.

BACKGROUND: With 9.6 million deaths in 2018, cancer remains the second leading cause of death worldwide. Breast cancer is the most deadly type of cancer among females, with 55.2% of crude incidence rate and 16.6% of crude mortality rate. PURPOSE: The present study was aimed to investigate the anti-breast cancer potential of natural dietary flavonoid, apigenin isolated from Clerodendrum viscosum leaves. METHODS: Apigenin was evaluated for in-depth anticancer activity in MCF-7 cells using cell viability assay, cell cycle analysis, Annexin-V-FLUOS staining, ROS induction, morphological analysis, and western blot analysis. RESULTS: Apigenin showed selective cytotoxicity on MCF-7 cells with an IC50-56.72 ± 2.35 µM, while negligible cytotoxicity was observed on WI-38 cells. Further, the flow cytometer-based analysis showed that apigenin halted MCF-7 cells in the G2/M phase arrest followed by dose-dependent apoptosis. Moreover, the FACS and confocal microscopy results confirmed the elevation of intracellular ROS and nuclear fragmentation in apigenin-treated MCF-7 cells. Western blots showed up-regulation of cell cycle regulatory proteins, increased p53 expression, Bax/Bcl-2 ratio, activation of caspases, and cleavage of PARP. Finally, apigenin treatment in the presence of Pifithrin-µ showed decreased apoptotic population and it was further confirmed through western blotting study. The results revealed the vital role of p53 in apigenin-induced apoptosis in MCF-7 cells. CONCLUSIONS: In the present findings, treatment of apigenin-induced intracellular ROS in MCF-7 cells followed by induction of G2/M phase cell cycle arrest and further apoptosis through the regulation of p53 and caspase-cascade signaling pathway.

African Vegetables (Clerodendrum volibile Leaf and Irvingia gabonensis Seed Extracts) Effectively Mitigate Trastuzumab-Induced Cardiotoxicity in Wistar Rats.

Trastuzumab (TZM) is a humanized monoclonal antibody that has been approved for the clinical management of HER2-positive metastatic breast and gastric cancers but its use is limited by its cumulative dose and off-target cardiotoxicity. Unfortunately, till date, there is no approved antidote to this off-target toxicity. Therefore, an acute study was designed at investigating the protective potential and mechanism(s) of CVE and IGE in TZM-induced cardiotoxicity utilizing cardiac enzyme and oxidative stress markers and histopathological endpoints. 400 mg/kg/day CVE and IGE dissolved in 5% DMSO in sterile water were investigated in Wistar rats injected with 2.25 mg/kg/day/i.p. route of TZM for 7 days, using serum cTnI and LDH, complete lipid profile, cardiac tissue oxidative stress markers assays, and histopathological examination of TZM-intoxicated heart tissue. Results showed that 400 mg/kg/day CVE and IGE profoundly attenuated increases in the serum cTnI and LDH levels but caused no significant alterations in the serum lipids and weight gain pattern in the treated rats. CVE and IGE profoundly attenuated alterations in the cardiac tissue oxidative stress markers' activities while improving TZM-associated cardiac histological lesions. These results suggest that CVE and IGE could be mediating its cardioprotection via antioxidant, free radical scavenging, and antithrombotic mechanisms, thus, highlighting the therapeutic potentials of CVE and IGE in the management of TZM-mediated cardiotoxicity.

Green Synthesis of Gold and Silver Nanoparticles Using Leaf Extract of Clerodendrum inerme; Characterization, Antimicrobial, and Antioxidant Activities.

Due to their versatile applications, gold (Au) and silver (Ag) nanoparticles (NPs) have been synthesized by many approaches, including green processes using plant extracts for reducing metal ions. In this work, we propose to use plant extract with active biomedical components for NPs synthesis, aiming to obtain NPs inheriting the biomedical functions of the plants. By using leaves extract of Clerodendrum inerme (C. inerme) as both a reducing agent and a capping agent, we have synthesized gold (CI-Au) and silver (CI-Ag) NPs covered with biomedically active functional groups from C. inerme. The synthesized NPs were evaluated for different biological activities such as antibacterial and antimycotic against different pathogenic microbes (B. subtilis, S. aureus, Klebsiella, and E. coli) and (A. niger, T. harzianum, and A. flavus), respectively, using agar well diffusion assays. The antimicrobial propensity of NPs further assessed by reactive oxygen species (ROS) glutathione (GSH) and FTIR analysis. Biofilm inhibition activity was also carried out using colorimetric assays. The antioxidant and cytotoxic potential of CI-Au and CI-Ag NPs was determined using DPPH free radical scavenging and MTT assay, respectively. The CI-Au and CI-Ag NPs were demonstrated to have much better antioxidant in terms of %DPPH scavenging (75.85% ± 0.67% and 78.87% ± 0.19%), respectively. They exhibited excellent antibacterial, antimycotic, biofilm inhibition and cytotoxic performance against pathogenic microbes and MCF-7 cells compared to commercial Au and Ag NPs functionalized with dodecanethiol and PVP, respectively. The biocompatibility test further corroborated that CI-Ag and CI-Au NPs are more biocompatible at the concentration level of 1-50 µM. Hence, this work opens a new environmentally-friendly path for synthesizing nanomaterials inherited with enhanced and/or additional biomedical functionalities inherited from their herbal sources.

Anti-inflammatory properties of the ethanol extract from Clerodendrum cyrtophyllum Turcz based on in vitro and in vivo studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Clerodendrum cyrtophyllum Turcz, a plant belonging to the Verbenaceae family, has been used in traditional medicine for the treatment of various inflammatory diseases in many Asian countries. AIM OF THE STUDY: The study aimed to evaluate anti-inflammatory properties of the ethanol extract from Clerodendrum cyrthophyllum Turcz leaves (EE-CC) through in vitro and in vivo models. MATERIAL AND METHODS: Total phenolic and flavonoid contents in the extract were determined using colorimetric methods and HPTLC. In red blood cell membrane stabilization model, rat erythrocyte suspension was treated with crude ethanol extract at different concentrations, the hemoglobin content of the supernatant solution released by red blood hemolysis was estimated. We also evaluated the effects of the ethanol extract from this plant on the production of nitric oxide (NO), tumor necrosis factor alpha (TNF-α) in stimulated RAW 264.7 cells. In order to elucidate its anti-inflammatory molecular mechanisms, we further evaluated the effects of the EE-CC on the expression of the inflammatory genes in inflammation-induced zebrafish model by tail-cutting using qPCR analysis. RESULTS: Colorimetric methods and HPTLC revealed high phenolic and flavonoid contents in the extract. In the red blood cell membrane stabilization model, the amount of hemoglobin released by red blood hemolysis significantly decreased in the presence of EE-CC, demonstrating a strong membrane stabilizing activity. EE-CC did not cause any toxic effect on cell viability but strongly inhibited NO and TNF-ɑ release due to LPS induction. The association with EE-CC significantly reduced the expression of cox-2, pla2, c3a, il-1(il1fma), il-8 (cxcl8b.1), tnf-α, and nf-ƙb, while increased the expression of the anti-inflammatory cytokine il-10 gene in cut-tail induced inflammation of zebrafish model. CONCLUSIONS: Taken together, the results suggest that the raw ethanol extract from C. cyrtophyllum Turcz leaves presents potent anti-inflammatory activities and may be useful for the treatment of various inflammatory diseases.

Clerodendrum petasites S. Moore: The therapeutic potential of phytochemicals, hispidulin, vanillic acid, verbascoside, and apigenin.

Clerodendrum petasites S. Moore has been prescribed in Thai traditional medicine for over 30 years for the treatment of ailments including asthma, inflammation, fever, cough, vomiting, and skin disorders. The phytochemicals from this plant have been identified as phenolic acids, flavones, flavone glycosides, glycosides, phenylpropanoid, and diterpenoid. The pharmacological activities both in vitro and in vivo have mostly been reported from crude extracts and not from pure compounds. This review, therefore, brings together information on the specific phytochemicals found in C. petasites in order to provide a guide to the natural bioactive compounds that are potentially used in medicines together with mechanisms underlying their pharmacological uses. All relevant information was searched for the terms of plant name, naturally-occurring compounds, and traditional uses from reliable databases, such as PubMed, Science Direct and Google Scholar, along with Thai traditional medicine textbooks. There was no specific timeline set for the search and this review selected to report only mechanisms studied by using standard compounds for their biological activities. Four dominant compounds comprising hispidulin, vanillic acid, verbascoside, and apigenin, have robust evidence to support their medical effects. Hispidulin was discovered to be possibly responsible for the treatment of cancer, osteolytic bone diseases, and neurological diseases. Other compounds were also found to tentatively support the uses in inflammation and neurological diseases. C. petasites extracts may provide an option as complimentary medicine, and or for the pharmacological development of new drugs derived from the phytochemicals found within.

Enhanced antimicrobial, antioxidant, in vivo antitumor and in vitro anticancer effects against breast cancer cell line by green synthesized un-doped SnO2 and Co-doped SnO2 nanoparticles from Clerodendrum inerme.

A novel approach was employed for the synthesis of un-doped tinoxide and Cobalt-doped tinoxide (Co-doped SnO2) nanoparticles (NAPs) by using aqueous extract of Clerodendrum inerme with the help of eco-friendly superficial solution combustion method. Synthesized NAPs were characterized by different spectroscopic techniques and results from XRD, TEM, SEM, EDX and UV-Vis examines confirmed the successful synthesis, crystalline nature and spherical structure of un-doped SnO2 and Co-doped SnO2 NAPs with the average grain size of 30 and 40 nm; and band gap energy of 3.68 and 2.76 eV respectively. Antimicrobial propensity of the synthesized NAPs was determined by agar well assay, SEM, TEM and confocal laser scanning microscopic analysis against various bacterial and fungal strains. Synthesized Co-doped SnO2 NAPs were unveiled the extraordinary antibacterial and antifungal activities against E. coli, B. subtilis, A. niger, A. flavus, and C. albicans with the zone of inhibitions of 30 ±â€¯0.08 mm and 26 ±â€¯0.06 mm, 17 ±â€¯0.04 mm, 23 ±â€¯0.08 mm and 26 ±â€¯0.06 respectively which were also evidenced from SEM, TEM and confocal laser scanning microscopy. In addition, green synthesized Co-doped SnO2 NAPs were demonstrated the substantial antioxidant activity by scavenging DPPH, significant in vitro anticancer and in vivo antitumor activity on breast carcinoma cells (MCF-7) and Ehrlich ascites tumor cell lines respectively than standard. The hemolytic activity disclosed low cytotoxicity of fabricated NAPs (0.89 ±â€¯0.05%) at 5 mg/mL, which was indicated their biocompatibility potential. Hence, the multi-purpose properties of synthesized NAPs presented in the current study can be further deliberated for pharmaceutical and nanomedicine applications.

Attenuation of oxidative stress and hepatic damage by white butterfly (Clerodendrum volubile) leaves in streptozotocin-induced diabetes in rats.

Background The negative impact of diabetes on the liver is well documented. The white butterfly (Clerodendrum volubile) leaf has been used in traditional practices for the treatment of various diseases, such as hypertension, diabetes, and rheumatism, but without scientific validation. This work was designed to evaluate the hepatoprotective properties of Clerodendrum volubile leaves on oxidative stress in streptozotocin (STZ)-induced diabetes in rats. Methods The rats were divided into ten groups of five rats each. Diabetes was induced by a single injection of STZ (65 mg/kg body weight; i.p), while the C. volubile extract (at the respective doses of 50, 100 and 200 mg/kg body weight) was given to diabetic and non-diabetic rats orally for 14 days. Metformin (100 mg/kg body weight) served as the positive control. Biochemical assays were conducted on the plasma for hematological parameters, along with hepatic marker damages and antioxidant enzyme determination in vivo to assess hepatic injury. Results The diabetic control rats showed significant increase (p<0.05) in marker enzymes: aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and malondiadehyde with reduction in reduced glutathione, glutathione peroxidase, superoxide dismutase, catalase activities and plasma total protein content. Likewise, there were alterations in hematological indices in the diabetic control rats when compared with the normal control. However, treatment with C. volubile caused a reversal of the above parameters towards normal levels, thereby suggesting the modulating effect of the extract on oxidative stress, which may be a result of the high polyphenolic content and antioxidant capacity. Conclusions The protection of the liver tissues and the modulation of oxidative stress in STZ diabetic rats compare favorably to metformin, a standard antidiabetic drug.

Clerodendrum volubile inhibits key enzymes linked to type 2 diabetes but induces cytotoxicity in human embryonic kidney (HEK293) cells via exacerbated oxidative stress and proinflammation.

The toxicity and safety associated with the use of medicinal plants remains a major concern. In this study, the antidiabetic properties of the dichloromethane (DCM) fraction of C. volubile leaves were investigated in vitro. Its cytotoxic effect and mechanism of toxicity were also investigated in Human Embryonic Kidney (HEK293) cells. The fraction was subjected to in vitro antioxidant assays using the 2,2'-diphenyl-1-picrylhydrazyl (DPPH) scavenging and Ferric reducing antioxidant power (FRAP) protocols. Its enzyme-inhibitory properties were investigated on α-glucosidase and α-amylase activities. Gas Chromatography Mass Spectroscopy (GCMS) and Fourier Transform Infrared (FTIR) spectroscopic analysis were used to identify its phytoconstituents. Cytotoxicity was determined via MTT assay. The treated cells were assayed for reduced glutathione (GSH), non-protein thiol, nitric oxide and malondialdehyde (MDA) levels, as well as Superoxide Dismutase (SOD), catalase, myeloperoxidase and ATPase activities. Cell apoptosis and/or morphological changes were determined using the acridine orange and ethidium bromide (AO/EB) dual staining method. The fraction showed significant (p < 0.05) antioxidant and enzyme-inhibitory activity. It showed significant (p < 0.05) cytotoxic effect against HEK293 cells with concomitant depletion of antioxidative and elevation of proinflammatory biomarkers. Morphological changes were examined in the cells with an apoptotic index of 0.84. 1,1-Dodecanediol, diacetate was identified as the most predominant compound, while aromatics and amines as the most functional groups present in the fraction. These results suggest the antidiabetic and cytotoxic effects of C. volubile leaves. The toxicity can be attributed to induced oxidative stress and proinflammation with concomitant depletion of ATP leading to apoptosis of the cells.