Cloning, bioinformatics, expression mode analysis of crocetin glycosyltransferase UGTCs4 / 中草药

Objective: With the aim to obtain crocetin glycosyltransferase UGTCs4 in cultured saffron suspension cell, and carry out its bioinformatics and expression mode analysis. Methods: A homologous cloning strategy and 5’ RACE methods were adopted, the full-length cDNA sequence of a crocetin glycosyltransferase, designated UGTCs4 (GeneBank number: KX398932), was obtained. The characteristics of physiochemical properties, structure and function of the deduced UGTCs4 protein were determined using a series of bioinformatics tools. Semi-quantitative PCR was used for gene expression analysis. Results: The results showed that the full length cDNA of UGTCs4 was 1 380 bp in length and encoding a 459 amino acid; UGTCs4 had high identities (83.2%) with UGTCs2 protein from saffron; UTGTCs4 had the same evolutionary tree as UGTCs2. UGTCs4 transcripts were constitutively expressed in the leaves, stems, and roots. UGTCs4 gene could respond to multiple treatments of indoleacetic acid (IAA), abscisic acid (ABA), gibberellin (GA), hydrogen peroxide (H2O2), and methyl jasmonate (MJA), which promoted its transcription. Conclusion: cDNA of crocetin glycosyltransferase was cloned from suspension cells for the first time and the response of UGTCs4 to different inducers was confirmed. Molecular characterization of UGTCs4 will be useful for further functional determination of the gene involving in the crocin biosynthesis and expression regulation.

Research progress on pharmacokinetics and dosage forms of crocin and crocetin / 中草药

Crocin is derived from stigmas of Crocus sativus and fruits of Gardenia jasminoides. It is a type of unique water-soluble carotenoids with long conjugate, possessing remarkable neuroprotection, cardiovascular protection, antitumor, and other pharmacological activities. Data showed that orally administrated crocin couldn’t permeate into the bloodstream as the prototype, but it can be absorbed into the blood circulation after being hydrolyzed to crocetin in gastrointestinal tract. Then, crocetin is converted to glucuronic acid conjugate. Moreover, the metabolite crocetin are mainly distributed in heart, liver, spleen, lung, and other blood-rich tissues. In view of the low bioavailability and poor stability of crocin and its metabolite crocetin, researchers have extensively employed novel drug delivery system to study crocin (crocetin) in order to improve its stability, bioavailability, and pharmacological activity. This article reviews in vivo metabolism of crocin (crocetin) in recent years and investigations on drug delivery system, therefore, facilitating further research and development of these natural products.

Variations of Chemical Constituents in Gardeniae Fructus Before and After Stir-frying by LCMS-IT-TOF / 中国实验方剂学杂志

Objective: To rapidly analyze chemical compositions in raw products and stir-fried products of Gardeniae Fructus,and determine the pharmacodynamic material basis of Gardeniae Fructus Praeparatus. Method: Liquid chromatography-ion trap-time-of-flight-mass spectrometry(LCMS-IT-TOF) was performed on ACQUITY UPLC HSS T3 column(2.1 mm×100 mm,1.8 μm),mobile phase was 0.005%formic acid aqueous solution(A)-acetonitrile(B) for gradient elution(0-5 min,10%-15%B;5-8 min,15%B;8-18 min,15%-95%B;18-20 min,95%B;20-22 min,95%-10%B;22-25 min,10%B),the column temperature was 30℃ and the flow rate was 0.4 mL·min-1,volume of sample injection was 2 μL;electrospray ionization(ESI) was applied for mass spectrometric analysis under positive and negative ion mode,the scanning ranges were m/z 100-1 000.The ion peaks were identified by comparison of control substances,mass spectrometry data analysis and literature references.The peak areas of some ion peaks before and after processing were compared,the changes of chemical compositions in Gardeniae Fructus after processing were investigated. Result: Based on the mass spectral data information and references,a total of 38 compounds were identified,no new compounds were produced after processing.The ion peak areas of main compounds were investigated during the stir-frying processing,the contents of geniposide,genipin-1-β-D-gentiobioside,shanzhiside,chlorogenic acid and crocin in Gardeniae Fructus Praeparatus were decreased;the contents of geniposidic acid and crocetin were increased. Conclusion: The method can rapidly and accurately identify the chemical constituents in Gardeniae Fructus Praeparatus.The changes of iridoids and crocins in Gardeniae Fructus during the stir-frying process may be related to hemostatic activity and protection of cardiovascular and cerebrovascular.

Crocetin promotes autophagy in injured rat hepatocytes induced by lipopolysaccharide and D-galactosamine / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of crocetin on autophagy in rat hepatocytes exposed to lipopolysaccharide (LPS) and D-galactosamine (D-gal) and explore the mechanism.</p><p><b>METHODS</b>Cultured rat hepatocytes were exposed to LPS (1 mg/L) and Dgal (60 mg/L) to induce cell injury and treated with crocetin, 3MA, or crocetin+3MA. Twelve hours after the treatments, the cells were examined for levels of ALT, AST and LDH in the supernatant using ELISA. LC3 fluorescence in the cells following immunofluorescence staining was observed using fluorescence microscopy. Autophagosomes in the cells were observed by transmission electron microscopy, and the cellular expressions of LC3, p62 and SIRT1 were detected using Western blotting.</p><p><b>RESULTS</b>The levels of ALT, AST and LDH in the hepatocytes were elevated after LPS- and D-gal-induced injury, reached the highest levels after 3MA treatment, but were decreased significantly by crocetin treatment. LC3 fluorescence increased obviously in the injured hepatoctyes, and the increment was the most obvious in crocetin-treated cells; LC3 fluorescence was decreased significantly after 3MA treatment. Cell injury induced obvious increase in autophagy in the hepatocytes, and the number of autophagosomes increased significantly after crocetin treatment but was reduced significantly after 3MA treatment. The cell injury caused an obvious up-regulation of LC3 and SIRT1 expression and down-regulated p62 expression. LC3 and SIRT1 expression levels were the highest and the expression of p62 was the lowest in cells with crocetin treatment. 3MA treatment significantly reduced the expression of LC3 and SIRT1 and increased the expression of p62 in the injured cells.</p><p><b>CONCLUSIONS</b>Autophagy is increased in injured rat hepatocytes, and crocetin can promote autophagy in the injured cells to reduce further cell injury.</p>

Research progress on biosynthetic pathway of apo-carotenoids in Crocus sativus / 中草药

The apo-carotenoid compounds represented by crocins are the main medicinal components of Crocus sativus, which have extensive anti-oxidation, anti-inflammatory, anti-atherosclerosis, anticancer, antidepressant, and other pharmacological activities. Biosynthetic pathways of apo-carotenoids in C. sativus include the traditional upstream route of the synthesis of geranylgeranyl pyrophosphate to zeaxanthin starting from mevalonate, and downstream pathway for the specific synthesis of crocetin and crocin by cleavage of zeaxanthin. This article reviews the recent research of key enzymes involved in the metabolism of apo-carotenoids in C. sativus, which facilitates further analysis of downstream pathways for the synthesis of apo-carotenoid derivatives such as crocin, and further provides a theoretical basis for the use of metabolic engineering methods to increase the production of crocins and other pharmacodynamic substances.

Analysis on transitional components in blood after oral administration of saffron extracts in rats / 中草药

Objective: To analyze the transitional components in blood after oral administration of saffron extracts in rats, in order to initially reveal in vivo potential effective substance and metabolic process of the main components absorbed into blood. Methods: The constituents of saffron and metabolites in rat plasma sample after oral administration of saffron were investigated using HPLC-DAD-ESI-MSn. Results: Eleven chemical constituents in saffron were identified, and four compounds, including one prototype and three metabolites, were detected in rat plasma sample. Among them, picrocrocin was absorbed in prototype, whereas crocins were absorbed in metabolites (crocetin and crocetin-monoglucuronide). Crocin-I and crocin-II, used as quality evaluation of saffron in Chinese Pharmacopoeia, were not discovered at all time points. Picrocrocin achieved a Cmax value at 1 h after administration, while metabolites, crocetin, achieved two Cmax values respectively at 0.5 h and 2 h. Conclusion: Picrocrocin and metabolic products absorbed into blood may be the effective components of saffron. The work may provide a reference for reasonably choosing the quality control index and improving the quality control standards, and may also lay a foundation for further researching the pharmacokinetics of saffron.

Effects of Crocetin on the Pupillary Response during Accommodation Induced by Visual Display Terminal Work: A Randomized, Double-blind, Placebo-controlled, Crossover Trial / 日本補完代替医療学会誌

The purpose of this study was to examine the effects of crocetin on the pupillary response during accommodation induced by visual display terminal (VDT) work. This clinical study was conducted as a randomized, double-blind, placebo-controlled, crossover trial in healthy adult volunteers with subjective symptoms of eye fatigue. In this study, the pupillary response during accommodation was evaluated using pupil constriction ratio (PCR). PCR was measured before and after VDT work, after rest at baseline, and at each intervention period. Following analysis of variations in PCR, the variation in PCR after rest significantly increased in the crocetin group. According to the visual analog scale questionnaire, subjective symptoms of eye fatigue significantly improved. These results show that ingestion of crocetin for 4 weeks is effective in mitigating the pupillary response during accommodation associated with VDT work.

Liver protection of crocetin against paraquat poisoning in rats / 中华危重病急救医学

Objective To study the liver protection of crocetin against paraquat (PQ) poisoning induced acute liver injury in rats. Methods Fifty-four male Wistar rats were randomly divided into control group, exposure group and treatment group, and the rats in each group were subdivided into the 0.5th, 2nd, and 6th day after exposure subgroups (n = 6). The model of acute liver failure induced by PQ poisoning was reproduced by intraperitoneal injection of 20 mg/kg of 20% PQ, and the rats in control group was injected with the same amount of normal saline. The rats in treatment group were given with intraperitoneal injection of 50 mg/kg crocetin after 0.5 day, once a day until they were sacrificed; the other two groups were injected with the same amount of normal saline. The rats in all groups were sacrificed at the corresponding time points, and blood was collected from inferior vena cava and hepatic tissue was harvested. Hematoxylin and eosin (HE) staining was used to observe the pathological changes in liver tissue on the 6th day under light microscope. Enzyme linked immunosorbent assay (ELISA) was used to detect the serum tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expressions of inducible nitric oxide synthase (iNOS) and nuclear factor-κB (NF-κB). The activities of apoptosis related factors, including caspase-8, -9, -12, in hepatic tissue were determined on the 6th day with chromogenic substrate method. Results In the liver tissue of exposed group, extensive infiltration of the inflammatory cells and the diffuse fragments necrosis were visible, and the regeneration of the liver cells was not obvious, and severity of the injury in a time dependent way. In the treatment group, the structure of hepatic artery was visible, and the infiltration of necrosis, congestion and inflammatory cells were not obvious. On the 0.5th, 2nd, and 6th day, serum levels of IL-6 and TNF-α, the mRNA expressions of iNOS and NF-κB in liver tissue, and the caspase-8, -9, -12 activities on the 6th day in the exposure group and treatment group were significantly higher than those in the control group. And the parameters in treatment group were significantly lower than those of the exposure group [IL-6 (ng/L): 188.37±64.21 vs. 376.61±82.42 on the 0.5th day, 287.18±58.69 vs. 432.77±96.28 on the 2nd day, 234.24±10.17 vs. 375.41±37.59 on the 6th day; TNF-α (ng/L): 472.36±76.43 vs. 688.33±102.19 on the 0.5th day, 189.32±87.54 vs. 296.21±89.77 on the 2nd day, 99.28±16.13 vs. 168.41±66.78 on the 6th day; iNOS mRNA (gray value): 2.998±0.801 vs. 3.453±0.026 on the 0.5th day, 3.126±0.306 vs. 5.259±0.153 on the 2nd day, 0.841±0.135 vs. 1.225±0.057 on the 6th day; NF-κB mRNA (gray value): 1.569±0.818 vs. 2.361±0.063 on the 0.5th day, 2.345±0.489 vs. 4.668±0.368 on the 2nd day, 2.348±0.316 vs. 3.972±0.449 on the 6th day; caspase-8 (pmol/mg): 126.77±9.97 vs. 199.18±66.48 on the 6th day; caspase-9 (pmol/mg): 213.12±69.06 vs. 321.62±89.39 on the 6th day; caspase-12 (pmol/mg): 183.46±70.52 vs. 219.68±53.93 on the 6th day, all P < 0.05]. Conclusion Crocetin has protective effect on liver in rats with PQ poisoning, which role is related with reducing the blood level of inflammatory factors, inhibiting the hepatic caspase-8, -9, -12 activities and gene expressions of iNOS and NF-κB.

Comparative Evaluation of Hepatoprotective Activities of Geniposide, Crocins and Crocetin by CCl4-Induced liver Injury in Mice

Iridoid glycosides (mainly geniposide) and crocetin derivatives (crocins) are the two major active constituents in Gardenia jasminoides Ellis. In the present study, geniposide, crocins, crocin-1 and crocetin were separated from gardenia chromatographically. Then, mice were orally administrated with geniposide (400 mg/kg b.w.), crocins (400 mg/kg b.w.), crocin-1 (400 mg/kg b.w.) and crocetin (140 mg/kg b.w.) once daily for 7 days with CCl4. Hepatoprotective properties were evaluated by biochemical parameters: Administration of geniposide, crocins, crocin-1and crocetin significantly lowered serum alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) levels in CCl4-treated mice. The reduced glutathione (GSH) levels and antioxidant enzymes (SOD and CAT) activities were also increased by geniposide, crocins, crocin-1 and crocetin. Histopathological examination of livers showed that these components reduced deformability, irregular arrangement and rupture of hepatocyte in CCl4-treated mice. These biochemical results and liver histopathological assessment demonstrated that geniposide, crocetin derivatives and crocetin show comparative beneficial effects on CCl4-induced liver damage via induction of antioxidant defense. Therefore, contents of geniposide and crocetin derivatives should be both considered for hepatoprotective efficacy of Gardenia jasminoides Ellis.

Antifibrotic effects of crocetin in scleroderma fibroblasts and in bleomycin-induced sclerotic mice

OBJECTIVE: To investigate the antifibrotic effects of crocetin in scleroderma fibroblasts and in sclerotic mice. METHODS: Skin fibroblasts that were isolated from three systemic scleroderma (SSc) patients and three healthy subjects were treated with crocetin (0.1, 1 or 10 μM). Cell proliferation was measured with an MTT assay. Alpha-smooth muscle actin was detected via an immunohistochemical method. Alpha 1 (I) procollagen (COL1A1), alpha 1 (III) procollagen (COL3A1), matrix metalloproteinase (MMP)-1 and tissue inhibitor of matrix metalloproteinase (TIMP)-1 mRNA levels were measured using real-time PCR. SSc mice were established by the subcutaneous injection of bleomycin. Crocetin (50 mg/kg/d) was injected intraperitoneally for 14 days. Dermal thickness and lung fibrosis were assessed with Masson's trichrome staining. Plasma ET-1 was detected with an enzyme-linked immunosorbent assay (ELISA). Skin and lung ET-1 and COL1A1 mRNA levels were measured via real-time PCR. RESULTS: Crocetin inhibited the proliferation of SSc and normal fibroblasts, an effect that increased with crocetin concentration and incubation time. Crocetin decreased the expression of α-SMA and the levels of mRNA for COL1A1, COL3A1 and matrix metalloproteinase-1, while crocetin increased TIMP-1 mRNA levels in both SSc and normal fibroblasts. Skin and lung fibrosis was induced, and the levels of ET-1 in the plasma, skin and lungs were elevated in bleomycin-injected mice. Crocetin alleviated the thickening of the dermis and lung fibrosis; decreased COL1A1 mRNA levels in the skin and lung; and simultaneously decreased ET-1 concentrations in the plasma and ET-1 mRNA levels in the skin and lungs of the bleomycin-induced sclerotic mice, especially during the early phase (weeks 1-3). CONCLUSION: Crocetin inhibits cell proliferation, differentiation and collagen production in SSc fibroblasts. Crocetin alleviates skin and lung fibrosis in a bleomycin-induced SSc ...

Pharmacokinetics of crocin-1 after oral administration in rats / 中国药学杂志

OBJECTIVE: To elucidate the intestinal absorption and pharmacokinetic characteristics of crocin-1, a major active constituent of the stigmas of saffron (Crocus sativus L) in rats after oral administration. METHODS: Extracted plasma samples were eluted by the mobile phase composed of methanol-2% acetate acid (pH 2) (48: 52 for crocin-1 and 74: 36 for crocetin) on Agilent Zorbax SB-C18 column monitored at 420 nm. RESULTS: An accurate, precise and selective analysis method was developed to determine crocein in rat plasma with the quantitation limit of 10 ng · mL-1. No crocin-1 was detected in both original and β-glucuronidase treated plasma samples. However, appreciable crocetin, the aglycon form of crocin-1, was observed in the original plasma. The concentration of crocetin did not increase obviously after hydrolization by β-glucuronidase. The average concentration-time data after oral administration of 1 mg · kg-1 crocin-1 showed that crocin-1 was rapidly absorbed as erocein and remained above 50 ng · mL-1 for about 10 h. The first peak appeared at 20 min and the second peak at 6 h. The area under curve (AUC), mean residence time (MRT) during 0 - 24 h and elimination half-life (t1/2k) were calculated as (1.16 ± 0.22) μg · h · mL-1, (5.4 ± 0.7) and (3.0 ± 0.6) h, respectively. CONCLUSION: Crocin-1 is absorbed as its hydrolyzation metabolite and shows relatively good pharmacokinetic characteristics. Crocetin may be thereby related to the pharmacological activities of crocin-1 in vivo. Copyright 2012 by the Chinese Pharmaceutical Association.

Comparative evaluation of the antioxidant capacity of crocetin and crocin in vivo / 中国药理学通报

Aim To investigate the antioxidant capacity of crocetin and crocin in an in vivo system.Methods Column chromatography was applied to the seperation of crocetin and crocin-1 from gardenia.Crocetin(6.25,12.5 and 25.0 mg·kg~(-1)·d~(-1)) and crocin (18.7,37.5 and 75.0 mg·kg~(-1)·d~(-1)) were orally administered to kunming mice.Then,superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),total antioxidant capacity(TAOC)and malondialdehyde(MDA)in mice were determined for the comparison of antioxidant activity of crocetin and crocin-1.Results Oral administration of crocetin and crocin for six weeks could enhance SOD of liver and kidney,GSH-Px of liver and TAOC of heart and kidney.In addition,it could decrease MDA of serum in mice.Conclusions The comparison of results suggests the evidence supporting the comparable antioxidant activity of crocetin and crocin.The results of the research also indicate that liver and kidney are two organs targeted for protection concerning endogenous antioxidant among various tissues.

Transgenic Tobacco Expressing the Crocus CSzCD Gene Showed the Production of Crocetin / 中国生物工程杂志

To investigate the feasibility to produce crocetin in tobacco plants.The coding region of zeaxanthin cleavage dioxygenase (CSzCD) gene from Crocus sativus L. was inserted into the downstream of the cauliflower mosaic virus (CaMV) 35S promoter of a binary vector pBI121, and integrated into the genome of Nicotiana tabacum L. Twenty-one transgenic lines were identified by genomic southern blotting analysis. Western blotting and HPLC analysis of the leaf extracts of transgenic tobacco showed that crocetin was produced in CSzCD gene-transformed plants, while no crocetin was found in the untransformed tobacco.

Effects of crocetin on doxorubicin-induced damage in rats with myocardial mitochondria / 中国临床药理学与治疗学

AIM: To study the protective effects of crocetin on the doxorubicin-induced damage in rats with myocardial mitochondria. METHODS: Rats were given intraperitoneal injection of doxorubicin to induce cardiotoxicity. After continuous oral administration of crocetin, the rats were sacrificed, and myocardial mitochondria were isolated. The mitochondrial membrane potential (MMP), O~-_2. production, the activity of respiratory chain complex IV and glutathione peroxidase (GSH-PX), and DNA fragmentation were determined. The expression level of COII gene was determined through RT-PCR. RESULTS: Crocetin increased the activity of respiratory chain complex IV and GSH-PX, MMP and the expression level of COII and decreased DNA fragmentation and superoxide anion radical O~-_2. production in rats with myocardial mitochondria. CONCLUSION: Crocetin may decrease the damage in rat myocardial mitochondria induced by doxorubicin.

Protective effects of crocetin on cerebral ischemia-reperfusion injury in mice / 中国临床药理学与治疗学

AIM: To research the protective effects of crocetin on cerebral ischemia-reperfusion(CIR) injury in mice.METHODS: The effects of crocetin on brain edema,brain capillary permeability,the activities of lactate dehydrogenase(LDH),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px) in brains and the contents of malondialdehyde(MDA) and nitric oxide(NO) in CIR model mice were observed.RESULTS: Compared with the CIR group,crocetin(50,100 mg/kg i.g.for 5 d) decreased the content of water,Evans blue,MDA and NO in mice brain tissue,and increased LDH、SOD and GSH-Px activity in brains of CIR mice,with statistical significance.CONCLUSION: Crocetin has protective effects on cerebral ischemia-reperfusion injury in mice,the mechanism may be related to its anti-oxidative activity and inhibition of NO overproduction.

Effects of crocetin on formation of advanced glycation end products and expression of releptor for advanced glycation and prodmts protein in diabetic rats / 中国临床药理学与治疗学

AIM: To study the effect of crocetin on the formation of advanced glycation end products(AGEs) and receptor for AGEs(RAGE) protein expression in diabetic rats induced by streptozotocin(STZ).METHODS: Rats were injected STZ in tail vessel with a dose of 45(mg?kg~(-1)).3 d later,the rats whose blood glucose were over(11.1)(mmol?L~(-1)) were regarded as diabetic rats,and were divided randomly into two groups: diabetic mellitus(DM) group,crocetin(50(mg?kg~(-1)),po) group.At the same time,8 normal rats were regarded as control group.After 21 d treatment,The levels of fasting blood glucose(FBG),serum fructosamine(FMN) and glycosylated hemoglobin(GHb) were measured.The contents of AGEs in aorta and mesenteric vessel were detected by fluorospectrophotometry.HE staining for mesenteric aorta was performed,and RAGE protein expression was studies with immunohistochemical method.RESULTS: Compared with DM group,crocetin did not decrease the level of FBG.However,it could decrease the levels of FMN and GHb in blood and AGEs in aorta and mesenteric vessel(P

Protection of crocetin on myocardial ischemia-reperfusion injury in rats / 中草药

Objective To study the protection of crocetin on myocardial ischemia-reperfusion injury and the possible mechanism involved. Methods Infarction model: Coronary occlusion was maintained for 45 min and reperfused for 180 min to produce infarction model. Creatine kinase (CK) and lactate dehydrogenase (LDH) activity in serum of infarction groups were analyzed at baseline (before ischemia), 5, 60, and 180 min after reperfusion. Stunned model: Stunned model was achieved by ligating the left anterior descending branch of coranary artery for 15 min. Na+, K+-ATPase, and Ca2+, Mg2+-ATPase activity, total adenosine content (TAC), and energy charge (EC) in ischemic and nonischaemic myocardial tissue and malondialdehyde (MDA) in serum of stunned groups were analyzed at the end of 180 min of reperfusion. Haemodynamic parameters including heart rate, left ventricular end-systolic and end-diastolic pressure, ejection fraction, maximum and minimum dp/dt, maximum and minimum dV/dt, and stroke work, were monitored at baseline, 5, 30, 90, and 180 min after reperfusion. Results Crocetin could attenuate the infarction ratio, relieve stunning in a dose-dependent manner, and decrease the CK and LDH activities in serum. Crocetin (50 mg/kg) could improve the heart function, ameliorate myocardial stunning degree, and increase the level of ATP and EC in myocardial tissue. Conclusion Crocetin could protect myocardium from ischemia and following reperfusion, then prevent the myocardial infarction and relieve myocardial stunning. The underlying mechanism may be partly attributed to the improvement of energy restoration and free radical scavenging effect.

Protection of crocetin on primary culture cardiac myocyte injured by hydroxyl free radicals / 中草药

Object To study the cardio-protective effect of crocetin on primary culture cardiac myocyte injured by hydroxyl free radicals. Methods After adding crocetin, primary culture cardiac myocyte was injured by 0.5 mmol/L hydroxyl free radicals which were generated by Fenton systems. The activity of lactic dehydrogenase (LDH) and mitochondrion succinic dehydrogenase (MSD) was observed. Mitochondrion membrane potential (MMP) was detected by Rh123. The ratio of cardiac myocyte apoptosis was investigated by flow cytometry and DNA ladder electrophoresis. Results Compared with the model group, crocetin significantly decreased the activity of LDH in culture supernatant and the ratio of cardiac myocyte apoptosis, markedly increased the activity of MSD and MMP. Conclusion Crocetin has the protective effect on the apoptosis of cardiac myocyte injured by hydroxyl free radical.

Effect of crocetin on ATPase and hydroxyproline in myocardial hypertrophy rat induced by overload pressure / 中草药

Object To study the influence of crocetin on the activity of ATPase and the content of hydroxyproline in the myocardial hypertrophy rats induced by overload pressure. Methods Abdominal aortic ligation was used to establish the myocardial hypertrophy model in rats. The rats were divided into two groups: 1) The early term group which included five groups: sham-operation group, model group, Captopril groups, crocetin 100, 50 mg/kg groups. 2) The long term group which also included five groups as the same as the early term groups. Heart indexes were examined. The ATPase activity of heart and the content of hydroxyproline in heart were assayed by colorimetric analysis. Results Compared with the model group, crocetin can significantly reduce the cardiac indexes, increase the activities of Na +, K +-ATPase; Ca 2+ , Mg 2+ -ATPase and markedly decrease the content of hydroxyproline in heart. Conclusion Crocetin could prevent the remodeling of cardiac hypertrophy induced by overload pressure. The ATPase may play an important role in myocardial hypertrophy induced by overload pressure.

Protective effect of crocetin on isoproterenol-induced myocardial injury in rats / 中草药

Object To study the protective effect of crocetin on myocardial ischemia induced by isoproterenol (ISO). Methods Myocardial ischemia was induced by subcutaneous injection of ISO (30 mg/kg for two days, once a day). The experimental animals were randomly divided into the normal group, ISO group, positive control group and three crocetin groups (25, 50, 100 mg/kg). Cardiac indexes were examined. The level of LDH, CK, MDA in serum were measured. The content of MDA, GSH-Px and the ATPase activity heart and mitochondria were assayed by colorimetric analysis. The histopathological change of myocardia was investigated by HE staining. Results Compared with the model group, crocetin can significantly reduce the cardiac indexes, obviously decrease the level of MDA, LDH and CK in the serum. Crocetin can significantly increase the activities of Na +, K +-ATPase; Ca 2+ , Mg 2+ -APTase; GSH-Px. The histopathological changes confirmed the protective effects of crocetin on the myocardial injury. Conclusion Crocetin could alleviate the acute myocardial ischemia induced by ISO in rats.