Crocin averts functional and structural rat hepatic disturbances induced by copper oxide nanoparticles.

Objective To observe the effect of Crocin on structure and the expression of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in rat retina after injury by ischemia-reperfusion.Methods A total of 80 Sprague-Dawley male rats at the age of 8-10 weeks were divided into control group,model group,low-dose Crocin group and high-dose Crocin group,with 20 rats in each group.The rats of control group were not treated.The rats in model,low-dose Crocin and high-dose Crocin group were induced with normal saline by anterior chamber perfusion creating a retinal ischemia-reperfusion (RIR) model.The rats of the low-dose Crocin and high-dose Crocin group received intraperitoneal injection with different doses of Crocin solution (5 mg/kg,or 50 mg/kg) 30 minutes prior to ischemic injury and one time per day after successful RIR.Optical microscopy was used to observe the retinal structure.Enzyme-linked immunosorbent assay (ELISA) was used to measure the expression of TNF-α and IL-1β 6,12,24 and 48 hours after RIR.Results The retinal structure of control group was normal.Pathological changes were found in the RIR model and low-dose Crocin group,such as retinal edema,disorganized structure and loosely packed cells.The degree of pathological changes in low dose Crocin group was less than the RIR model group.The retinal structure of high-dose Crocin group was similar to the control group.The expression of TNF-α was the highest at 24 hours after modeling,while the expression of IL-1β was the highest at 12 and 48 hours after RIR modeling.Six,12,24 and 48 hours after RIR modeling,compared with the control group,the TNF-α expression of model (t=5.42,7.94,9.32,9.18; P＜0.05),low-dose Crocin (t=3.94,4.12,4.98,3.84; P＜0.05) and high-dose Crocin group (t=2.13,2.34,2.96,2.78;P＞0.05) were increased.Compared with the RIR model group,the TNF-α expression of low-dose Crocin (t=3.95,4.56,4.01,5.12) and high-dose Crocin group (t =5.23,7.65,7.74,7.63) was decreased.Compared with the control group,the IL-1β expression of model (t=7.23,7.87,7.15,15.60),low-dose Crocin (t 5.65,5.10,5.54,6.87;P＜0.05) and high-dose Crocin group (t=4.38,5.21,4.56,4.75) was increased (P＜0.05).Compared with the model group,the IL-1β expression of low-dose Crocin group was decreased significantly 48 hours after RIR modeling (t=7.56,P＜0.05) ; but it decreased significantly at each time point in high-dose Crocin group (t=6.94,5.36,6.05,10.50; P＜0.05).Conclusion Crocin can improve the retinal pathologic changes,while down regulating TNF-α and IL-1β expression in RIR rats. Key words: Reperfusion injury/drug therapy; Colchicum/drug effects; Tumor necrosis factor alpha; Interleukin-1beta; Animal experimentation

Background The primary cause of glaucomatous optic nerve damage is apoptosis of nerve cell and disorder of retinal circulation.Erigeron breviscapus is confirmed to have a protective effect on retinal ganglion cells (RGCs) in rats with chronic ocular hypertension, and crocin has the effect of anti-inflammation and anti-apoptosis.However, whether crocin can protect RGCs against ocular hypertension damage is unclear. Objective This study was to investigate the protective effect of crocin on the optic nerve in chronic ocular hypertension. Methods Thirty-two SD rats were randomized into the sham operation group, model control group, erigeron group and crocin group, 8 rats for each group.The right eyes served as experimental eyes.Chronic ocular hypertensive models were established by episcleral vein cauterization in the rats of the model control group, erigeron group and crocin group, and only conjunctiva was cut off in the sham operation group.Erigeron (150 mg/kg) and crocin (20 mg/kg) was intraperitoneally injected in the erigeron group and crocin group respectively 30 minutes before operation and once daily after operation for 4 weeks, and 0.5 ml normal saline was used in the sham operation group and model control group.Intraocular pressure (IOP) was measured before surgery and 1 day, 3 days, 1 week, 2 weeks, 3 weeks and 4 weeks after surgery.The samples of eyeballs and optic nerve in the rats were prepared at 4 weeks after surgery.Retinal thickness was measured by hematoxylin and eosin staining.The apoptosis of RGCs was detected by TUNEL assay.The ultrastructure change of optic nerve was examined under the transmission electron microscope, and the expression of bcl-2 and bax proteins in retinal homogenates was analyzed by Western blot.The use and care of the animals followed the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Committee. Results The IOPs were significantly elevated in the model control group, erigeron group and crocin group in comparison with the sham operation group, and the IOPs was significantly higher in various time points after surgery than that before surgery (Fgroup=169.079, P=0.000; Ftime=50.505, P=0.000). The retinal thickness was (192.72±4.28), (165.15±3.89), (177.75±3.35) and (182.48±4.12)μm in the sham operation group, model control group, erigeron group and crocin group, and rat retinal thickness in the crocin group was significantly lower than that in the sham operation group and higher than that in model control group and erigeron group (all at P<0.05). The percentage of apoptotic RGCs in the sham operation group, model control group, erigeron group and crocin group, showed significant reduction in comparison with the model control group and erigeron group (all at P<0.05). The number of myelinated nerve fibers and bcl-2/bax values were significantly increased in the crocin group compared with the model control group and erigeron group (all at P<0.05). Conclusions Crocin affords a optic nerve by inhibiting RGC apoptosis and optic nerve degeneration in SD rats with chronic ocular hypertension, and the protecting effect of crocin is more prominent than that of erigeron breviscapus. Key words: Crocus/chemistry; Herbal medicine; Plant extracts/pharmacology; Ocular hypertension; Apoptosis; Retinal ganglion cells; Erigeron/chemistry; Optic nerve protection

Objective To investigate the protective effect of crocin on oxidative stress cell model of PC12 cell and the effect of crocin on PI3K/Akt signal pathway, as well as further explore the mechanism of protective effect on model cells. Methods Cells were divided into control group, model group, crocin group and VE group. The cell survival rate was detected by MTT method, and the expression of mRNA and protein of PI3K/Akt were detected by RT-PCR and Western blot. Results With the crocin concentration in 0.625 μM and 5 μM, the cell survival rate increased in a dose-dependent manner. The average optical density rate of PI3K and Akt mRNA were 0.435±0.044 and 0.375±0.034, and the PI3K and Akt protein were 0.378±0.038 and 0.386±0.043 of crocin group.Compared with the model group, the expression levels of PI3K/Akt increased in crocin group(P<0.05). Conclusion These results indicate that the antioxidant and antiapoptosis effects of crocin are induced via increasing expression of PI3K and pAkt. Key words: Crocin; Oxidative stress; Apoptosis; PI3K/AKT

Crocin treatment prevents doxorubicin-induced cardiotoxicity in rats

We examined the extent and severity of radiographic periodontal bone loss in patients with different stages of chronic kidney disease (CKD) and explored a potential dose-response relationship between bone loss and CKD-related biomarkers. Panoramic radiographs were obtained from 129 CKD patients (78 males and 51 females; mean age: 63.5 years, range: 24 to 91 years), including 63 patients undergoing dialysis for an average of 3.3 years (range: 0.5 to 14 years). Glomerular filtration rate (GFR), dialysis dose, and levels of serum biomarkers were obtained through a hospital database. Interproximal bone loss was assessed as a percentage of root length. Twenty-nine participants were edentulous (23.8% of those on dialysis versus 21.2% of those with residual kidney function; χ(2) test, P = 0.724). The extent of bone loss was higher among dialysis patients (analysis of variance [ANOVA], P = 0.007), but no clear dose-response association between CKD stage and extent was evident. GFR, dialysis dose, and levels of serum biomarkers did not differ between edentulous and dentate individuals, and only serum albumin was lower in patients with extensive bone loss (ANOVA, P = 0.030). After adjusting for dialysis status, the severity of bone loss was positively associated with glucose levels (multiple regression, P = 0.019) and white blood cell count (P = 0.032), whereas the number of teeth present was positively associated with plasma phosphorus (P = 0.008) and negatively with glucose levels (P = 0.011). Despite a higher extent of bone loss in dialysis patients, the lack of a dose-response association between bone loss and CKD stage or the levels of CKD-related serum biomarkers underscores the complex relationship between the two conditions.

Protective effects of crocin against gentamicin-induced damage in rat testicular tissue: Modulating the levels of NF-κB/TLR-4 and Bax/Bcl-2/caspase-3 signaling pathways.

The results showed that after treating &lt;italic&gt;C. albicans&lt;/italic&gt; with Artemisinin (0.104 mg/ml) for 3 hours the number of &lt;italic&gt;C. albicans&lt;/italic&gt; decreased and affected the morphology hyphae and growth of &lt;italic&gt;Candida&lt;/italic&gt; compared with Fluconazole (150 mg/ml) and control (infected mice). The biomarker levels in the tongue showed high levels of MDA and TGFβ (623±1.5pg/ml, 586.1±0.13pg/ml respectively) in (infected mice with fungi). IL-37 was recorded high level (49.21±0.21pg/ml) in (Neoral +Fungi + artemisinin + fluconazole) compared with negative control. The finding biomarker levels in stomach showed high levels of MDA and IL-37 (533.8±1.9, 69.76±0.39pg/ml) in (Neoral + Fungi + artemisinin + fluconazole) compared with control. TGF β was recorded high level (1002±0.32pg/ml) in (Neoral + Fungi + Artemisinin) compared with negative control. The biomarker levels in intestine showed high levels of MDA and TGFβ (1149±0.34pg/ml, 1089±0.3pg/ml respectively) in (mice infected with fungi). IL-37 was recorded high level (74.14±0.14pg/ml) in (Neoral + Fungi + artemisinin + fluconazole) compared with negative control (normal mice). The biomarker levels in serum showed high levels of MDA and TGFβ (1668±0.12 pg/ml, 1629±0.05 pg/ml respectively) in (mice infected with fungi) at (P&lt;0.05). IL-37 was recorded high level (135.1±0.8pg/ml) in (negative control). In conclusion, artemisinin has a role as antifungal and &lt;italic&gt;C. albicans&lt;/italic&gt; infection causes the imbalance in immunity.

The investigation of oxidant/antioxidant status in hepatic tissues from cholesterol-fed rabbits and the establishment of possible protective effects of aqueous garlic extract on cholesterol-induced hepatic steatosis. Twenty-two of 31 white New Zealand rabbits were given cholesterol (0.5 g/kg/day) for 4 months. Seven of them were then killed (cholesterol group). The remaining 15 animals were divided into two groups. Seven were fed on a normal laboratory diet (normal diet group) and the others (extract group) on a normal diet plus garlic extract (1.5 ml/kg/day) for an additional 3 months. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) enzyme activities, antioxidant potential (AOP) value, malondialdehyde (MDA), cholesterol and triglyceride levels in the liver tissues and total cholesterol and triglyceride levels in serum samples were measured. An histological evaluation was also done. An impaired antioxidant system, reduced antioxidant defence potential and increased peroxidation were found in hepatic steatotic tissues from cholesterol-fed animals. Treatment with garlic extract caused a significant increase in antioxidant potential and partly eliminated peroxidation damage in the hepatic tissue. Additionally, the extract caused significant reductions in the cholesterol levels of blood and hepatic tissues. The histological evaluations were in accordance with these results. The results suggest that cholesterol-induced steatosis leads to a weakened antioxidant defence system and causes peroxidation in the hepatic tissue. Treatment with garlic extract may contribute to significant amelioration in the hepatic steatosis and peroxidation processes.

<strong>Introduction</strong>: Muscular inflammation and atrophy is one of the characteristics of diabetes that causes motor disability in these individuals. Studies have shown that exercise training with different intensities and the use of herbal drugs can have favorable effects on diabetes. The aim of this study was to interactive effects of continuous and interval training with crocin consumption on interleukin 17 and 18 in the Soleus Muscle of type 2 Diabetic Rats. <strong>Methods</strong>: In this experimental study, 49 adult diabetic rats were randomly assigned to seven groups, including: high intensity interval training (HIIT), low intensity continuous training (LICT), HIIT + crocin consumption, LICT + crocin consumption, crocin consumption, sham, and control. HIIT and LICT groups did training for eight weeks on rodent treadmill, respectively. Crocin groups received 25 mg/kg of daily crocin for 8 weeks peritoneally. The gene expression levels of the variables were measured using Real Time-PCR. <strong>Results</strong>: HIIT increased IL-17 and 18 in the Soleus muscle tissue (p≤0.05), but LICT does not have a significant effect on changes in IL-17 and IL-18 (p≥0.05). Crocin consumption decreased expression of IL-18 and increased IL-17 in the Soleus muscle tissue (p≤0.05), and interaction of LICT and crocin consumption was significant in increasing IL-17 and IL-18 (P≤0.05). <strong>Conclusion</strong>: HIIT seems to have inflammatory effects in the muscle tissue of type 2 diabetic rats. However, the interaction of LICT and crocin was significant in the increase of IL-17 and IL-18 in the Soleus muscle tissue of type 2 diabetes rats.

Obesity is accompanied by adipose tissue remodeling characterized by increased production of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, leptin and resistin and reduced secretion of adiponectin, which favors inflammation, metabolic disorders, and cardiovascular diseases. Although intragastric balloon (IGB) can be considered safe and effective for weight loss, its effect on serum levels of these biomarkers has been evaluated only in a few studies, while no previous study evaluated its effect on circulating levels of resistin, TNF-α, and IL-6. The aim of this study was to evaluate the changes in serum levels of metabolic and inflammatory biomarkers in obese patients submitted to IGB treatment. A prospective observational study involving 42 patients with obesity using IGB for 6months. The patients were evaluated, on the day of insertion and withdrawal or adjustment of IGB, for the following: anthropometric measures and serum levels of adiponectin, leptin, resistin, TNF-α, IL-6, high-sensitivity C-reactive protein (hs-CRP), glucose, insulin, uric acid, triglycerides, and total cholesterol and fractions. The body mass index decreased from 35.15 ± 0.41 to 29.50 ± 0.54kg/m2. There was a reduction (p < 0.05) in leptin, hs-CRP, glucose, insulin, HOMA-IR, and triglycerides, while the adiponectin/leptin ratio increased (p < 0.05). Moreover, weight loss presented (1) a positive association with the decrease in leptin, hs-CRP, glucose, insulin, HOMA-IR, uric acid, and total cholesterol and (2) a negative association with the reduction in adiponectin/leptin ratio. The present study suggests that 6months of IGB treatment in obese individuals reduce serum leptin and hs-CRP and improves insulin resistance and lipid profile which may decrease cardiovascular risk.

Simple SummaryLiver cancer represents one of the most lethal forms of cancer, with hepatocellular carcinoma (HCC) accounting for the majority of its incidences and deaths. Currently, sorafenib is the first-in-line option for treating advanced and unresectable HCC. It is a multi-kinase inhibitor that intervenes with tumor growth and progression. Considering the modest results provided by sorafenib, identifying novel approaches to treating HCC remains a clinical imperative. Based on our previous work, crocin, a constituent of saffron, prevented HCC development. This study aimed to investigate its therapeutic effect in combination with sorafenib against an induced model of hepatocellular carcinoma arising from a cirrhotic milieu in rats. Our results confirmed that combination therapy yielded more effective outcomes compared to crocin and sorafenib monotherapies. It exerted the most pronounced effects in inhibiting inflammation and tumor cell proliferation while activating apoptosis and restoring macroscopic and cellular liver morphology. These results introduce a potential strategy for optimizing the anticancer effects of sorafenib using the bioactive natural compound crocin against HCC.Hepatocellular carcinoma (HCC) is one of the most aggressive malignancies, with continuously increasing cases and fatalities. Diagnosis often occurs in the advanced stages, confining patients to systemic therapies such as sorafenib. Sorafenib (SB), a multi-kinase inhibitor, has not yet demonstrated sufficient efficacy against advanced HCC. There is a strong argument in favor of studying its use in combination with other medications to optimize the therapeutic results. According to our earlier work, crocin (CR), a key bioactive component of saffron, hinders HCC development and liver cancer stemness. In this study, we investigated the therapeutic use of CR or its combination with SB in a cirrhotic rat model of HCC and evaluated how effectively SB and CR inhibited tumor growth in this model. Diethylnitrosamine (DEN) was administered intraperitoneally to rats once a week for 15 weeks, leading to cirrhosis, and then 19 weeks later, leading to multifocal HCC. After 16 weeks of cancer induction, CR (200 mg/kg daily) and SB (10 mg/kg daily) were given orally to rats for three weeks, either separately or in combination. Consistently, the combination treatment considerably decreased the incidence of dyschromatic nodules, nodule multiplicity, and dysplastic nodules when compared to the HCC group of single therapies. Combined therapy also caused the highest degree of apoptosis, along with decreased proliferating and β-catenin levels in the tumor tissues. Additionally, when rats received combined therapy with CR, it showed anti-inflammatory characteristics where nuclear factor kappa B (NF-κB) and cyclooxygenase-2 (Cox-2) were considerably and additively lowered. As a result, CR potentiates the suppressive effects of SB on tumor growth and provides the opportunity to strengthen the therapeutic effects of SB in the treatment of HCC.

Clinical use of triptolide (TP) is restricted due to severe toxicity. This study assessed the protective effect of crocin (CR) as a natural antioxidant against TP-induced toxicity in bovine collagen type II-induced arthritis (CIA) in mice. The mice in the CIA model group showed macroscopic signs of severe arthritis. The anti-arthritis effects in the control, TP + CR, and TP groups were evaluated through assessment of foot volume, arthritis score, and proinflammatory cytokines, and collagen antibody assay. Crocin reduced TP-induced toxicity, as evidenced by evaluation of survival rate, body weight, visceral index, hepatic and renal functions, histopathologic analyses, and antioxidant enzyme activities. Transcriptome sequencing resulted in identification of 76 differentially expressed genes (DEGs) associated with hepatotoxicity between the TP and TP + CR groups. Of these, Three DEGs (Cyp1a2,Gsta4, and Gstp1) were validated using quantitative real-time PCR analysis. In conclusion, CR protected CIA mice from TP-induced toxicity through modulation of the cytochrome P450 and glutathione metabolism pathways.

Copper oxide nanomaterials (CuO NPs) have been widely utilized in many fields, including antibacterial materials, anti-tumor, osteoporosis treatments, imaging, drug delivery, cosmetics, lubricants for metallic coating, the food industry, and electronics. Little is known about the potential risk to human health and ecosystems. The present work was conducted to investigate the ultrastructural changes induced by 20 ± 5nm CuO NPs in hepatic tissues. Adult healthy male Wister albino rats were exposed to 36 intraperitoneal (ip) injections of 25nm CuO NPs (2mg/kg bw). Liver biopsies from all rats under study were processed for transmission electron microscopy (TEM) processing and examination for hepatic ultrastructural alterations. The hepatic tissue of rats exposed to repeated administrations of CuO NPs exhibited the following ultrastructural alterations: extensive mitochondrial damage in the form of swelling, crystolysis and matrix lysis, formation of phagocytized bodies and myelin multilayer figures, lysosomal hyperplasia, cytoplasmic degeneration and vacuolation, fat globules precipitation, chromatin clumping, and nuclear envelope irregularity. The findings indicated that CuO NPs interact with the hepatic tissue components and could induce alterations in the hepatocytes with the mitochondria as the main target organelles of copper nanomaterials. More work is recommended for better understanding the pathogenesis of CuO NPs.

Crocin ameliorates methotrexate-induced liver injury via inhibition of oxidative stress and inflammation in rats.

To assess the diagnostic value of interleukin 6 (IL-6), IL-8 and tumor necrosis factor-α (TNF-α) as inflammatory markers in chronic obstructive pulmonary disease (COPD) patients. IL-6, IL-8 and TNF-α levels were measured by ELISA in the serum and the bronchoalveolar lavage (BAL) in 10 control participants and 25 mild and moderate COPD patients, whereas 25 patients with severe COPD were studied for the serum level of these inflammatory biomarkers. The mean value and SD of BAL and serum IL-6, IL-8 and TNF-α levels were significantly higher in COPD patients when compared with control participants; the serum level of these biomarkers were also significantly higher in severe compared with mild and moderate COPD patients. Increased srum and/or BAL IL-6, IL-8 and TNF-α can be used as biomarkers of the systemic inflammatory response in COPD patients, and their levels are correlated with the severity of COPD. Egypt J Broncho 2014 8:91–99