Evaluation of the effect of shodhana(detoxification)process using chromatographic profiling(HPTLC,HPLC,LC-MS,and GC-MS)and estimation of the content of toxic colchicine in Gloriosa superba Linn.tubers

Shodhana(purification/detoxification)is an important aspect of Ayurveda.It has been described in several important pieces of literature.In this study,we performed shodhana process on Langali(Gloriosa superba Linn.)tubers of family Liliaceae,a semi-woody herbaceous climber with significant therapeutic properties.We compared the unprocessed(before purification)and processed(after purification)samples based on the phytochemical,physicochemical,and chromatographic study,including HPTLC,HPLC,GC-MS and LC-MS.Significant changes were observed in profiling after the shodhana process.This plant contains colchicine,known for its toxic effects,particularly cardiotoxicity.Therefore,we quantified colchicine in the unprocessed and processed samples of Gloriosa superba Linn.tubers using HPLC method.Results showed that the percentage of colchicine was reduced by 13.67%in chloroform extract and 20.97%in ethanol extracts after the shodhana process.

Colchicine-induced Melilotus Research

[Objective] The aim was to identify the ploidy of sweet clover induced by colchicine. [Method] The sweet clover germinating seeds and colchicine solution were used for materials to research colchicine-induced Melilotus. [Result] The results showed that the concentration of 0.2% colchicine solution could induced double sweet clover which showed corresponding features of polyploidy plants as a whole showed great features. [Conclusion] Sweet clover induced by colchicine was polyploidy.

Colchicine reduces procollagen Ⅲ and increases pseudocholinesterase in chronic liver disease

AIM:To test whether colchicine would be an effective antif ibrotic agent for treatment of chronic liver diseases in patients who could not be treated with α-interferon.METHODS:Seventy-four patients(46 males,28 females) aged 40-66 years(mean 53±13 years) participated in the study.The patients were affected by chronic liver diseases with cirrhosis which was proven histologically(n=58);by chronic active hepatitis C(n=4),chronic active hepatitis B(n=2),and chronic persistent hepatitis C(n=6).In the four patients lacking histology,cirrhosis was diagnosed from anamnesis,serum laboratory tests,esophageal varices and ascites.Patients were assigned to colchicine(1 mg/d) or standard treatment as control in a randomized,double-blind trial,and followed for 4.4 years with clinical and laboratory evaluation.RESULTS:Survival at the end of the study was 94.6% in the colchicine group and 78.4% in the control group(P=0.001).Serum N-terminal peptide of type Ⅲ procollagen levels fell from 34.0 to 18.3 ng/mL(P=0.0001),and pseudocholinesterase levels rose from 4.900 to 5.610 mU/mL(P=0.0001) in the colchicine group,while no signif icant change was seen in controls.Best results were obtained in patients with chronic hepatitis C and in alcoholic cirrhotics.CONCLUSION:Colchicine is an effective and safe antifibrotic drug for long-term treatment of chronic liver disease in which fi brosis progresses towards cirrhosis.

Tolerance of nestin^+ cholinergic neurons in the basal forebrain against colchicine-induced cytotoxicity

In the present study we injected colchicine into the lateral ventricle of Sprague-Dawley rats to investigate the effects of colchicine on the number of different-type neurons in the basal forebrain and to search for neurons resistant to injury. After colchicine injection, the number of nestin^＋ cholinergic neurons was decreased at 1 day, but increased at 3 days and peaked at 14-28 days. The quantity of nestincholinergic neurons, parvalbumin-positive neurons and choline acetyl transferase-positive neurons decreased gradually. Our results indicate that nestin^＋ cholinergic neurons possess better tolerance to colchicine-induced neurotoxicity.

Extraction and Anti-inflammatory Effects of Colchicine from Sagittaria sagittifolia

[Objective] This study was to study the optimal extraction technology and anti-inflammatory effects of colchicine from Sagittaria sagittifolia. [Method] The ef- fects of ethanol concentration, extraction time, extraction temperature and solid-liquid ratio on the extraction rate of colchicine from S. sagittifolia were investigated. On the basis of single-factor experiments, an L9 （34） orthogonal test was carried out to optimize the extraction process. According to the optimal extraction process, the content of colchicine in S. sagittifolia was determined by high performance liquid chromatography. The anti-inflammatory ability of colchicine was evaluated through an anti-inflammatory test in vitro. [Result] The optimal extraction process of colchicine from S. sagittifolia was as follows： ethanol concentration of 60%, extraction temper- ature of 50℃, extraction time of 30 min, and solid-liquid ratio of 1：25 （g/ml）. The content of colchicine in S. sagittifolia was determined as 40.58 μg/100 mg. Com- pared with the control, the colchicine from S. sagittifolia （9.0 and 4.5 g/kg） inhibited the increase in PGE2, TNF-α and IL-1β contents in pleural fluid （P〈0.05）. High-dose colchicine inhibited the increase in TNF-α, IL-1β and MDA contents in lung tissue （P〈0.01）, and middle-dose colchicine inhibited the increase in IL-1β content in lung tissue （P〈0.01）. [Conclusion] The colchicine in S. sagittifolia has a good anti-inflam- matory effect, which may be achieved through hindering the production of inflam- matory mediators and antioxidation.

Development of a novel rat model of heterogeneous hepatic injury by injection with colchicine via the splenic vein

AIM To develop a novel rat model of heterogeneous hepatic injury.METHODS Seventy male Sprague-Dawley rats were randomly divided into a control group(n = 10) and a colchicine group(n =60). A 0.25% colchicine solution(0.4 mL/kg) was injected via the splenic vein in the colchicine group to develop a rat model of heterogeneous hepatic injury. An equal volume of normal saline was injected via the splenic vein in the control group. At days 3, 7, and 14 and weeks 4, 8, and 12 after the operation, at least seven rats of the colchicine group were selected randomly for magnetic resonance imaging(MRI) examinations, and then they were euthanized. Ten rats of the control group underwent MRI examinations at the same time points, and then were euthanized at week 12. T2-weighted images(T2 WI) and diffusion weighted imaging(DWI) were used to evaluate the heterogeneous hepatic injury. The heterogeneous injury between the left and right hepatic lobes was assessed on liver sections according to the histological scoring criteria, and correlated with the results of MRI study. RESULTS Obvious pathological changes occurred in the hepatic parenchyma in the colchicine group. Hepatic injury scores were significantly different between the left and right lobes at each time point(P < 0.05). There was a significant difference in apparent diffusion coefficient(ADC) of DWI and liver-to-muscle ratio(LMR) of T2 WI between the left and right lobes of rats in the colchicine group(P < 0.05) at each time point, and similar results were observed between the colchicine and control groups. Besides, there was a significant correlation between hepatic injury scores and ADC values or LMR(r =-0.682, P = 0.000; r =-0.245, P = 0.018).CONCLUSION Injection with colchicine via the splenic vein can be used to successfully develop a rat model of heterogeneous hepatic injury. DWI and T2 WI may help evaluate the heterogeneous injury among liver lobes.

Acute and recurring pericarditis: More colchicine,less corticosteroids

Acute and recurring pericarditis are frequently encountered clinical entities.Given that severe complications such as tamponade and constrictive pericarditis occur rarely,the majority of patients suffering from acute pericarditis will have a benign clinical course.However,pericarditis recurrence,with its painful symptoms,is frequent.In effect,recent studies have demonstrated a beneficial role of colchicine in preventing recurrence,while also suggesting an increase in recurrences with the use of corticosteroids,the traditional first-line agent.

Colchicine sensitizes human hepatocellular carcinoma cells to damages caused by radiation

AIM： We studied the effect of colchicine combined with radiation on the survival of human hepatocellular carcinoma （HCC） HA22T/VGH cells.METHODS： Twenty-four hours after treatment with 0-8 ng/mL colchicine, HA22T/VGH cells were irradiated at various doses （0, 1, 2, 4, and 8 Gy）. Colony assay was performed to assess the surviving cell fraction. Survival curves were fitted by using a linear-quadratic model to estimate the sensitizer enhancement ratio （SER）. Flow cytometry was used for cell cycle analysis.RESULTS： Colchicine at lower concentrations （1 and 2 ng/mL） had obvious synergy with radiation to inhibit HCC cell growth, whereas higher concentrations （4 and 8 ng/mL） had only additive effect to radiation. Pretreatment with 1 and 2 ng/mL colchicine for 24-h enhanced cell killing by radiation with SERs of 1.21 and 1.53, respectively.G2/N arrest was only observed with higher colchicine doses （8 and 16 ng/mL） after 24-h treatment, this effect was neither seen with lower doses （1, 2, and 4 ng/mL）nor with any dose after only 1 h of treatment.CONCLUSION： Our results suggest that colchicine has potential as an adjunct to radiotherapy for HCC treatment.Lower doses of colchicine possess radiosensitizing effects via some mechanism other than G2/M arrest. Further study is necessary to elucidate the mechanism.

Colchicine Inhibited the Expression of Tissue Inhibitor of Metalloprotenase-1 and Interleukin-6 in Cultured Activated Hepatic Stellate Cells

Cultured HSCs were treated colchicine with different concentrations for 12 h, respectively. The effects of eolehicine on HSCs growth were measured by MTT assay. Effects of eolchicine on gene expression of HSCs were analysed by using a self-made oligonucleotide mieroarray. Colehieine inhibited HSCs growth in a dose-dependent manner. After 12 h of treatment with 6.25 mg/L of colehicine, the expression of tissue inhibitor of metalloprotenase-1 （TIMP-1） and the expression of interleukin-6 （IL-6） in HSCs were downregulated by 2.3 folds and 2.1 folds, respectively. These results suggest that colchieine＇s beneficial effects may, at least in part, owe to the inhibitory to the proliferation of HSCs and down regulation of the expression of both TIMP1 and IL-6 in HSCs.

Induced Polyploidy via Colchicine Treatment Increases Flower Size and Fruit Weight in Cape Gooseberry(Physalis peruviana L.)

An induced polyploid plant through colchicine treatment offers probably the best scope for improvement in flower size and fruit weight.Thus,in the present study,an attempt was made to induce polyploidy in Cape gooseberry using colchicine with the objective of creating more genetic variability.The colchicine concentrations were used as 0.10%(C1),0.20%(C2)and 0.40%(C3)for the duration 12(H1),24(H2)and 36(H3)hours for each concentration with seedling apex dip method(M1),cotton plug method(M2)and lanolin paste method(M3).The plants treated with 0.10%of colchicine by cotton plug method for 12 h showed the better performance during the years 2017-2018 and 2018-2019 in respect of more delay in the flower bud emergence(54 d and 53 d from the date of transplanting),anthesis(19 d and 20 d from the first appearance of bud to full anthesis of flower)and fruit setting(8.00 d and 9.00 d from the date of anthesis),bigger flower size(2.93 cm2 and 3.00 cm2)than the untreated plants.The lower percentage of pollen viability(40.33%and 40.67%)was noticed in the same treatment in comparison to control(70.33%and 72.33%).The fruit maturity was also extended(59 d and 60 d from the date of fruit set)with bigger sized fruits(length:2.53 cm and 2.57 cm,breadth:2.27 cm and 2.33 cm)as well as more fruit weight(8.70 g and 8.33 g)by the application of colchicine at 0.10%with cotton plug method for 12 h.On the basis of results obtained,the application of colchicine at 0.10%for 12 h duration with cotton plug method was found to be the best and effective treatment for induction of polyploidy as well as more flower size and fruit weight in Cape gooseberry.

A Novel Approach for Production of Colchicine as a Plant Secondary Metabolite by in Vitro Plant Cell and Tissue Cultures

The secondary metabolites synthesized by plants are economically important chemical compounds in the agricultural and industrial areas such as food, perfumery and pharmaceutical sectors. In recent years, attempts for their production by in vitro plant cell and tissue cultures have been accelerated considerably. Colchicine, the principle secondary metabolite of Colchicum autumnale L. and Gloriosa superba L., is an important alkaloid that has poison effect used for treatment of various diseases and plant breeding studies. Presently, colchicine has been produced by using the seeds of C. autumnale L. and the tubers of G. superba L. through different chemical extraction methods. Applying in vitro plant cell and tissue cultures together with metabolic and genetic engineering techniques, large-scale production of colchicine can be achieved from the above two plant species.

Effects of Colchicine on Distribution of Mitochondria in 2-cell and Compacted 8-cell Mouse Embryos

The distribution of mitochondria during early development of mouse embryos was visualized bymitochondria-specific vital fluorescent dye, rhodamine 123(Rh 123). Mitochondrial clusters wasmarkedly conceotrated to perinuclear area in blastomere of normal 2-ccll embryos. In blastomere ofuncompacted 8-cell embryos, mitochondria were randomly distributed throughout the cytoplasm, butthey were reorganizcd to the cytocortices beneath the apposed surfaces of blastomere duringcompaction. As demonstrated in our study, colchicine (10 μg/ml) produced marked effect onmitochondrial distribution in blastomcre of 2-cell and compacted 8-cell embryos: mitochondriabecame scattered throughout the cytoplasm ofblastomere. It is suggested that the spatial distributionof mitochondria in early mouse embryo are maintained by microtubule.

SOFT X-RAY IMAGES OF CHROMOSOMES WITHIN INTACT TUMOR CELLS TREATED WITH COLCHICINE

In order to avoid any possible effect of separating procedures from intact cell on morphologic structure of the chromosomes, cultivated Hep-2 tumor cells were treated with colchicine and observed by soft X-ray contact microscopy for the first time. The fine structures of chromosomes are more clear with stereo features. The thread-like and coarse granular structures twine and tangle up together within chromosome masses which can not easily be revealed in Wright’s stained sample by light microscope or osmium stained sample by transmission electron

Genome-wide analysis of microRNAs induced and regulated by colchicine in mulberry

A variety of plants were colchicine treated to double their chromosome number.Chromosomes are genetic carriers that determine the individual traits of organisms.The doubling of chromosomes will lead to modifications in plant morphology,physiology and genetics.To determine the response of mulberry trees induced by colchicine,using mulberry variety Yu-711 leaves as research materials,two small RNA libraries(control and experimental groups)were constructed.It was found that 45 known miRNA genes and 78 predicted novel miRNA genes in the sequence results.A comparison of data between the control group and the experimental group revealed 37 differentially expressed miRNA genes,19 genes of which were up-regulated and 18 genes of which were down-regulated.Eight miRNAs were selected from 37 differentially expressed miRNA genes.These miRNAs were verified by quantitative real-time PCR,and the expression levels of these miRNAs were found to be consistent with those obtained by sequencing,which proved the accuracy of sequencing results.The PsRNATarget software was used to predict the target genes of 8 miRNAs.The Gene Ontology and Kyoto Encycopedia of Genes and Genomes analysis were used to collect the functions of target genes and confirm that target genes are mainly involved in biological processes,cell components and molecular functions.novel-77 miRNA was selected from 37 differentially expressed miRNAs.What will serve as a fundamental data in understanding mulberry miRNAs function in molecular biology research and further provides the molecular mechanism of mulberry gene regulation as induced by colchicine is that the prediction of novel_77 precursor sequence and the target gene(XM_010104258.2),as well as the secondary structure analysis.

Chronic colchicine poisoning with neuromyopathy,gastric ulcers and myelosuppression in a gout patient:A case report

Colchicine has been widely used as an anti-gout medication over the past decades.However,it is less commonly used due to its narrow therapeutic range,meaning that its lethal dose is close to its therapeutic dose.The lethal dose of colchicine is considered to be 0.8 mg/kg.As chronic colchicine poisoning has multiple manifestations,it poses a challenge in the clinician’s differential diagnosis.Historically,the drug was important in treating gout;however,clinical studies are currently underway regarding the use of colchicine in patients with coronavirus disease 2019 as well as its use in coronary artery disease,making this drug more important in clinical practice.CASE SUMMARY A 61-year-old male with a history of gout and chronic colchicine intake was admitted to our Emergency Department due to numbness and weakness of the lower limbs.The patient reported a history of colchicine intake for 23 years.After thorough examination,he was diagnosed with colchicine poisoning,manifesting as neuromyopathy,multiple gastric ulcers and myelosuppression.We advised him to stop taking colchicine and drinking alcohol.We also provided a prescription of lansoprazole and mecobalamin,and then asked him to return to the clinic for re-examination.The patient was followed up for 3-mo during which time his gout symptoms were controlled to the point where he was asymptomatic.CONCLUSION Colchicine overdose can mimic the clinical manifestations of several conditions.Physicians easily pay attention to the disease while ignoring the cause of the disease.Thus,the patient’s medication history should never be ignored.

Effect of Colchicine on Inducible Nitric Oxide Synthase Activity and Nitric Oxide Production of Mice Induced by <i>Aggregatibacter actinomycetemcomitans</i>

Objective: Colchicine induced a non-protective Th2-like immunity in Aggregatibacter actinomycetemcomitans-stimulated murine immune response. The aim of the present study was to determine whether colchicine affects inducible nitric oxide synthase (iNOS) activity and nitric oxide (NO) production in A. actinomycetemcomitans-immunized mice. Materials and Methods: BALB/c mice were sham-immunized (group I) or immunized with heat-killed A. actinomycetemcomitans (group II-VII). Colchicine was injected intraperitoneally before (group III), on the same day of (group IV), or after (group V) the primary immunization and on the same day of (group VI) or after (group VII) the secondary immunization. In vitro, spleen cells from either sham- or heat-killed A. actinomycetemcomitan-immunized animals were cultured and stimulated with heat-killed A. actinomycetemcomitans in the presence or absence of colchicine with or without addition of L-arginine, Db-cAMP, forskolin or interferon-γ (IFN-γ). The levels of splenic iNOS activity and both serum and culture supernatant NO levels were assessed. Results: The results showed that colchicine did inhibit both splenic iNOS activity and serum NO levels only when the drug was injected at the same time as the immunization (group IV and VI). Splenic iNOS activity and NO levels on antigen-stimulated spleen cell cultures were also suppressed by colchicine, even in the presence of L-arginine, Db-AMP or forskolin. IFN-γ only partially restored iNOS activity and NO levels in the antigen and colchicine-treated spleen cell cultures. Conclusion: This study suggests, therefore, that colchicine may suppress the iNOS activity and NO production in A. actinomycetemcomitans-immunized mice in vivo and in vitro.

Improving Growth Performance of Jatropha curcas by Inducing Polyploidy through Colchicine Treatment

This study was planned to assess genetic variation in Jatropha curcas as affected by colchichine treatments. During the experiment, seeds of Jatropha curcas were collected from mature and healthy Jatropha plants and were soaked in different concentrations of colchicines. The second treatment was that of time of soaking (seeds were soaked in different concentrations for different time durations). The results clearly revealed that by increasing the concentration of colchicine treatment, parameters like stomatal density, and size of the guard cells were reduced.

Colchicine-Induced Rhabdomyolysis and Possible Amiodarone Interaction ——Colchicine-Induced Rhabdomyolysis

Objective: To report a case of drug interaction leading to rhabdomyolysis. Case Summary: A 65-year old woman suf-fering from chronic atrial fibrillation was treated with amiodarone and acenocoumarol. Two weeks after administration of conventional dosage of colchicine for pericarditis, the patient developed rhabdomyolysis. colchicine-induced rhabdomyolysis was suspected. Colchicine was stopped and the patient underwent supportive therapy. Clinical symptoms improved rapidly. Discussion: Colchicine-induced neuromuscular toxicity and rhabdomyolysis have been reported with chronic treatment in therapeutic doses. Concomitant use of several drugs with colchicine may potentiate the development of myopathy. In our case, a co-administration of colchicine, a well known substrate of cytochrome P450 3A4 and P-glycoprotein, and amiodarone had possibly precipitated rhabdomyolysis. Amiodarone may increase colchicine toxicity by a dual mechanism. Amiodarone inhibits P-glycoprotein which may theoretically result in increased intrace- llular colchicine concentrations and decreased hepatic and renal excretion of the drug. Conclusion: Amiodarone may potentiate the development of colchicine-induced rhabdomyolysis.

Severe Colchicine Poisoning: A Case Report

Background: Colchicine intoxication is uncommon but potentially life threatening because of the high toxicity of the drug. Colchicine binds to the intracellular protein tubulin, causing disturbed mitosis in all tissues followed by multi-organ failure. Case Presentation: We report a case of a 23-year-old woman admitted to emergency room 12 hours after a suicide attempt by ingestion of approximatively 40 mg of colchicine. Few hours after consumption, first symptoms were a hemodynamic shock and ARDS, leading to her death due to multi-organ failure. Conclusion: The colchicine intoxication has strong morbidity for doses higher than 0.5 mg/kg. The treatment is symptomatic. The immunotherapy using anti-colchicine allows improving critical form’s prognosis.

Effect of Colchicine on Total Antioxidant Capacity,Antioxidant Enzymes and Oxidative Stress Markers in Patients with Knee Osteoarthritis

Background/Aim: We aimed to investigate the effects of colchicine on clinical recovery, as well as oxidative stress markers and total antioxidant capacity (TAC) in whole blood of patients with knee osteoarthritis (OA). Materials and methods: Sixty patients with grade 2 - 3 knee OA according to ACR knee OA criteria whom examination of the knee joint synovial fluid by polarized light microscopy demonstrated CPDD crystals existence were included in the study. Fifty healthy subjects were included as a control group. Patients were divided randomly into two groups. The first group (paracetamol group) was given only paracetamol 3 gr daily p.o and the second group (colchicine and paracetamol group) was given colchicine 1,5 gr and paracetamol 3 gr daily p.o for six months. For outcome measures WOMAC and VAS were used. Superoxide dismutase (SOD), Catalase (CAT) ezyme activities and Glutathione (GSH) and Malondialdehyde (MDA) levels and TAC all were measured. Results: WOMAC scores were improved in both patient groups compared with pre-treatment evaluation (p 0.05). TAC was signifcantly increased only in colchicine/paracetamol group. Oxidant parameter MDA levels were significantly decreased in both paracetamol group and colchicine/paracetamol group. CAT, SOD enzyme activities and GSH levels did not change before and after treatment protocols in both patient groups. Conclusion: Both paracetamol 3000 mg/day and 3000 mg paracetamol plus 1,5 gr/day colchicine is effective in the treatment of patients with knee osteoarthritis. But the addition of colchicine to paracetamol produced significantly greater symptomatic benefit than paracetamol alone. Our study also showed that colchicine lowers whole blood MDA which is a lipid peroxidation compound and elevates TAC levels in patients with knee OA. This may show probable disease modifying effect of colchicine in knee OA which require further long period laboratory and radiologic investigations.