Proliferating Cell Nuclear Antigen Labeling Research Articles

741. EXPRESSION OF COX2 AND P53 IN RAT ESOPHAGEAL CANCER INDUCED BY REFLUX OF DUODENAL CONTENTS

Abstract Aim. Reflux of duodenal contents can induce mucosal injury, stimulate cell proliferation, and promote tumorigenesis. We examined the expression of COX2 and p53 in rat esophageal lesions induced by duodenal content reflux. Methods. Thirty 8-week-old male Wistar rats were exposed to duodenal content esophageal reflux. All animals underwent an esophagoduodenal anastomosis (EDA) with total gastrectomy in order to produce chronic esophagitis. Ten rats were the sham. Control. They were sacrificed at the 40th week. Their esophagi were examined for HE, COX2, p53, and proliferating cell nuclear antigen (PCNA). Results. After 40 weeks of reflux, dysplasia, squamous cell carcinoma (SCC), and adenocarcinoma (ADC) were found. PCNA labeling index was higher in dysplastic and cancer tissue than that in normal. Overexpression of COX2 was shown in ADC and SCC. Wild-type p53 accumulation was found in ADC, and not in SCC. Conclusion. Reflux of duodenal contents into the esophagus led to ADC and SCC in rats. COX2 may play an important role in esophageal cancer by duodenal content reflux. Our present results suggest an association between wild-type p53 accumulation and COX2 expression in ADC, with no such relation seen in SCC.

Comparative Analysis of Cluster of Differentiation 57 and Proliferating Cell Nuclear Antigen Expression in Different Grades of Oral Squamous Cell Carcinoma: An Immunohistochemical Study.

The immune defense against tumor cells is mainly mediated by the natural killer (NK) cells. Cluster of differentiation 57 (CD57) is a 110-kd glycoprotein, typically expressed by the NK cells, attacks the cancer cells and inhibits tumor development. Proliferating cell nuclear antigen (PCNA) is a 36-kd auxiliary protein for DNA polymerase delta that correlates with cell proliferation and DNA synthesis. It is an essential component of DNA replication, DNA recombination, and DNA repair. The uncoordinated proliferation of PCNA protein characterizes the biological behavior of malignant lesions. The aim of the present study is to compare and correlate the expression of CD57 and PCNAin different grades of oral squamous cell carcinoma (OSCC) by immunohistochemistry. This retrospective analysis comprises 30 samples of various grades of OSCCsand10 samples of healthy mucosa.Sections of 4-5 µm thickness were done and stained with monoclonal anti-PCNA and anti-CD57 antibodies.The statistical package for social science (SPSS) version 16.0 software (IBM Corp., Armonk, NY) was used to analyze the data in this study. The expression of CD57 and PCNA was compared and correlated between the groups using analysis of variance (ANOVA) post hoc, Dunnet t-test, and Pearson's correlation coefficient test.For statistical significance, a p-value of 0.05 or less was used. A significant decrease in CD57 labeling index was seen from well-differentiated squamous cell carcinoma (16.63 ± 2.33) to poorly differentiated squamous cell carcinoma (5.53 ± 1.20) whereas the significant increase in PCNA labeling index was noted from well-differentiatedsquamous cell carcinoma (45.88 ± 2.20), followed by moderately differentiated and poorly differentiated squamous cell carcinoma (72.77 ± 4.35). The combination of CD57 and PCNA biomarkers appears to be good indicators of the immune status of the patient and the aggressiveness of the lesion.

18. REFLUX OF DUODENAL CONTENTS INDUCES ESOPHAGEAL CARCINOGENESIS (SCC, ADC)

Abstract The incidence of esophageal cancer patients who have undergone distal gastrectomy is increasing recently. Distal gastrectomy is a good model for studying the clinical effects of duodenal content reflux. It is known that reflux of duodenal contents can induce mucosal injury, stimulate cell proliferation, and promote tumorigenesis. We examined the expression of COX2 and P53 in rat esophageal lesions induced by duodenal contents reflux after total gasytrectomy. Thirty 8 week old male wistar rats were exposed to duodenal content esophageal reflux. All animal underwent an esophagoduodenal anastomosis(EDA) with total gastrectomy in order to produce chronic esophagitis. Ten rats were the sham (Control). They were sacrified at the 40th week. Their esophagi were examined for HE, COX2,P53 and Proliferating cell nuclear antigen (PCNA). After 40 weeks of reflux, dysplasia(100%), squamous cell carcinoma(SCC)(40%) and adenocarcinoma (ADC) (30%)were found. PCNA Labeling index was higher in dysplastic and cancer tissue than that of normal. Overexpression of COX2 were shown in ADC and SCC. Wild type p53 accumulation were found in ADC, not in SCC. Moreover, SCC developed in places distant from the anastomosis compared to ADC. As compared with ADC, SCC is less frequently inflammatory. This means that histological features may depend on the volume of reflux contents; small amounts of reflux causes SCC and a large volume of reflux causes ADC. Reflux of duodenal contents into the esophagus led to ADC and SCC in rat. Small amounts of reflux causes SCC and a large volume of reflux causes ADC. COX2 may play an important role in esophageal cancer by duodenal content reflux. Our present results suggest an association between wild type P53 accumulation and COX2 expression in ADC, with no such relation seen in SCC.

Proliferating Cell Nuclear Antigen (PCNA) Localization in the Ovaries of Buffalo, Dog and Pig by Immunohistochemistry

Background: It was expected from the previous studies that the immune histochemical staining for PCNA within follicular cells altered depending on the stage of follicular growth. Therefore, we aimed at determining the expression of proliferating cell nuclear antigen protein (PCNA) in the buffalo, dog and pig ovaries. Methods: The localization of PCNA was demonstrated in formalin fixed, paraffin embedded tissue sections of buffalo, dog and pig ovaries using primary mouse monoclonal anti-PCNA antibody. Result: Staining for PCNA was not observed in pregranulosa cells but observed in oocytes of primordial follicles. In primary to early secondary follicles, in oocytes and in few granulosa cells the positive staining was detected. The late secondary preantral follicles and actively growing small to large antral follicles showed strong PCNA labelling in the layers of granulosa and theca cells. PCNA staining was expressed in nuclei of oocytes in preantral and small antral follicles. The follicles demonstrating advanced atresia showed mild to no PCNA labelled granulosa and theca cells. The results of our study demonstrated that follicular growth and development in ovaries may be effectively monitored by determining the rate of granulosa cell proliferation of PCNA.

Long-Term Nonclinical Pulmonary Safety Assessment of Afrezza, a Novel Insulin Inhalation Powder.

Afrezza delivers inhaled insulin using the Gen2 inhaler for the treatment of patients with type 1 and type 2 Diabetes. Afrezza was evaluated in long-term nonclinical pulmonary safety studies in 2 toxicology species. Chronic inhalation toxicology studies in rat (26 weeks) and dog (39 weeks) and an inhalation carcinogenicity study in rats were conducted with Technosphere insulin (Afrezza) and with Technosphere alone as a vehicle control. Respiratory tract tissues were evaluated by histopathology and cells expressing proliferating cell nuclear antigen (PCNA) were quantified in lungs of rats. Microscopic findings in rats exposed to Afrezza were attributed to the Technosphere particle component, were confined to nasal epithelia, and consisted of eosinophilic globules and nasal epithelial degeneration. There were no Afrezza-related changes in pulmonary PCNA labeling indices in alveoli, large bronchioles, or terminal bronchioles. Microscopic findings in rats exposed to Technosphere particles included eosinophilic globules, mucus cell hyperplasia, and epithelial degeneration in the nasal cavities. PCNA labeling indices were increased in large bronchioles and terminal bronchioles but not in alveoli. There were no Technosphere particle-related findings in the dog study. Afrezza did not exhibit carcinogenic potential in the 2-year study in rats. These nonclinical inhalation studies support the use of Afrezza in humans over extended periods.

Thalidomide Reduces Cell Proliferation in Endometriosis Experimentally Induced in Rats.

To analyze the effect of thalidomide on the progression of endometriotic lesions experimentally induced in rats and to characterize the pattern of cell proliferation by immunohistochemical Proliferating Cell Nuclear Antigen (PCNA) labeling of eutopic and ectopic endometrium. Fifteen female Wistar rats underwent laparotomy for endometriosis induction by resection of one uterine horn, isolation of the endometrium and fixation of a tissue segment to the pelvic peritoneum. Four weeks after, the animals were divided into 3 groups: control (I), 10mg/kg/day (II) and 1mg/kg/day (III) intraperitoneal thalidomide for 10 days. The lesion was excised together with the opposite uterine horn for endometrial gland and stroma analysis. Eutopic and ectopic endometrial tissue was submitted to immunohistochemistry for analysis of cell proliferation by PCNA labeling and the cell proliferation index (CPI) was calculated as the number of labeled cells per 1,000 cells. Group I showed a mean CPI of 0.248 ± 0.0513 in the gland and of 0.178 ± 0.046 in the stroma. In contrast, Groups II and III showed a significantly lower CPI, that is, 0.088 ± 0.009 and 0.080 ± 0.021 for the gland (p < 0.001) and 0.0945 ± 0.0066 and 0.075 ± 0.018 for the stroma (p < 0.001), respectively. Also, the mean lesion area of Group I was 69.2 mm2, a significantly higher value compared with Group II (49.4 mm2, p = 0.023) and Group III (48.6 mm2, p = 0.006). No significant difference was observed between Groups II and III. Thalidomide proved to be effective in reducing the lesion area and CPI of the experimental endometriosis implants both at the dose of 1 mg/kg/day and at the dose of 10 mg/kg/day.

The Effect of Estrogen on Hepatic Fat Accumulation during Early Phase of Liver Regeneration after Partial Hepatectomy in Rats.

Fatty liver is common in men and post-menopausal women, suggesting that estrogen may be involved in liver lipid metabolism. The aim of this study is to be clear the role of estrogen and estrogen receptor alpha (ERα) in fat accumulation during liver regeneration using the 70% partial hepatectomy (PHX) model in male, female, ovariectomized (OVX) and E2-treated OVX (OVX-E2) rats. Liver tissues were sampled at 0–48 hr after PHX and fat accumulation, fatty acid translocase (FAT/CD36), sterol regulatory element-binding protein (SREBP1c), peroxisome proliferator-activated receptor α (PPARα), proliferative cell nuclear antigen (PCNA) and ERα were examined by Oil Red O, qRT-PCR and immunohistochemistry, respectively. Hepatic fat accumulation was abundant in female and OVX-E2 compared to male and OVX rats. FAT/CD36 expression was observed in female, OVX and OVX-E2 at 0–12 hr after PHX, but not in male rats. At 0 hr, SREBP1c and PPARα were elevated in female and male rats, respectively, but were decreased after PHX in all rats. The PCNA labeling index reached a maximum at 36 hr and 48 hr in OVX-E2 and OVX rats, respectively. ERα expression in OVX-E2 was higher than OVX at 0–36 hr after PHX. In conclusion, these results indicated that estrogen and ERα might play an important role in fat accumulation related to FAT/CD36 during early phase of rat liver regeneration.

The Toxic Effect Of Sildenafil Citrate On Adult Albino Rat Testis And The Possible Protective Role Of Royal Jelly (Histological And Immunohistochemical Study)

Background: Phosphodiesterase inhibitor was appearing as a magic tool in treatment of erectile dysfunction, however, it might lead to harmful effects in high dose and long term administration. The aim of the work: This study aimed to evaluate the toxic effect of sildenafil citrate on the histological structure of the testis of adult albino rats and the possible protective role of royal jelly. Materials and methods: Thirty five adult male albino rats were divided into 3 groups; the control group divided into (subgroup Ca, Cb and Cc), experimental group divided into (subgroup Ea and Eb) and treated group divided into (subgroup Ta and Tb). In the experimental group sildenafil citrate was given at a dose of 9 mg/kg in subgroup Ea and 13.5mg/kg in subgroup Eb orally daily for 30 days. In the treated group rats were treated by royal jelly at a dose of 100 mg/kg for 30 days orally after stoppage of sildenafil citrate. In the present study, the sperm count, histological and immunohistochemical studies were evaluated. Result : Daily oral administration of sildenafil citrate (9mg/kg and13.5 mg/kg ) for 30 day had reduced the sperm count and induced apparent degeneration with detached, distorted germ cell and congested blood vessels with significant decrease in Proliferating cell nuclear antigen Labeling Index (PCNA- LI). After royal jelly administration, there was an improvement in sperm count and histopathological changes induced by sildenafil citrate with a significant increase in PCNA -LI. Conclusion: Long-term daily use of Sildenafil citrate produced histological changes in the testis and reduced the spermatogenesis with a significant decrease in sperm count and these side effects can be prevented by using royal jelly.

Immunohistochemical Study of the Ameliorative Effect of Vitamin E on Liver Regeneration after Different Periods of Partial Hepatectomy

The liver is almost unique in its capacity for regeneration after hepatectomy but the exact mechanisms are not yet fully clarified. Antioxidants have been shown to promote liver regeneration after major hepatectomy. The present study evaluated the ameliorative effect of vitamin E administration on the liver regeneration after different periods of partial hepatectomy (PH) in rats. Fifty-six adult male albino rats were divided into three groups: Control sham operated group; partially hepatectomized group which were divided into three subgroups sacrificed at 1day, 3 days and 7days after the operation respectively; Partially Hepatectomized group with vitamin E pretreatment before PH where the rats were given a daily oral dose of vitamin E until the time of sacrifice of the rats. Immunohistochemical detection of proliferating cell nuclear antigen (PCNA) and labeling index were demonstrated. After PH, the PCNA positive hepatocytes and the PCNA labeling indices were significantly high after the 1st day and then much decreased after the 3rd day, to be followed by a slight increase at the 7th day. Vitamin E pretreatment in PH rats resulted in a decrease in PCNA positive cells and its labeling indices in the 1st day with a gradual increase in the 3rd and 7th days. Vitamin E has an inhibitory effect in the first 24 hours on liver regeneration followed by stimulatory effect at the third and seventh days after PH. These data indicated that vitamin E pretreatment has an important role in regulation and enhancement of liver regeneration after PH.

Inhibition of Formation of Azoxymethane-induced Colonic Aberrant Crypt Foci in Rats by Edible Green Algae Capsosiphon fulvescens and Brown Algae Hizikia fusiforme.

Capsosiphon fulvescens (green seaweed) and Hizikia fusiforme (brown seaweed) are marine algae consumed as food supplements, especially in Japan, China and Korea, and are considered traditional medicinal tonics for certain ailments. The aim of this study was to investigate the possible inhibitory effects of dietary C. fulvescens and H. fusiforme on azoxymethane (AOM)-induced colorectal cancer (CRC) in rats. F344 male rats (5 weeks, 150 g) were divided into six groups as follows. Group 1: Injected with normal saline solution and fed control diet (untreated control). Group 2: Injected with AOM and fed control diet (treated control). Group 3: Injected with AOM and fed 1% C. fulvescens diet. Group 4: Injected with AOM and fed 2% C. fulvescens diet. Group 5: Injected with AOM and fed 2% H. fusiforme diet. Group 6: Injected with AOM and fed 6% H. fusiforme diet. Test animals received subcutaneous injections of AOM (15 mg/1 ml/kg body weight) once a week for 2 weeks to induce aberrant crypt foci (ACF) in treated control and experimental groups. We evaluated the effects of dietary C. fulvescens and H. fusiforme at two different dose levels: 1 and 2% C. fulvescens, and 2 and 6% H. fusiforme, on colonic carcinogenesis by AOM in rats. Our results suggest that body weights were not significantly different amongst groups. We found that feeding C. fulvescens and H. fusiforme with a control diet significantly (p<0.05) inhibited the development of ACF in experimental groups. C. fulvescens and H. fusiforme in food also significantly (p<0.05) reduced the proliferating cell nuclear antigen labeling index in the colonic tissues of experimental groups. These results demonstrate the chemopreventive potential of C. fulvescens and H. fusiforme against CRC in an AOM-induced rats.

Reduced triglyceride accumulation due to overactivation of farnesoid X receptor signaling contributes to impaired liver regeneration following 50% hepatectomy in extra‑cholestatic liver tissue.

The aim of the present study was to investigate the role of triglyceride metabolism in the effect of obstructive cholestasis on liver regeneration following 50% partial hepatectomy (PH). Obstructive cholestatic rat models were achieved via ligation of the common bile duct (BDL). Following comparisons between hepatic pathological alterations with patients with perihilar cholangiocarcinoma, rats in the 7day post‑BDL group were selected as the BDL model for subsequent experiments. Liver weight restoration, proliferating cell nuclear antigen labeling index, cytokine and growth factor expression levels, and hepatic triglyceride content were evaluated to analyze liver regeneration post‑PH within BDL and control group rats. The results of the present study revealed that obstructive cholestasis impaired liver mass restoration, which occurred via inhibition of early stage hepatocyte proliferation. In addition, reduced triglyceride content and inhibited expression of fatty acid β‑oxidation‑associated genes, peroxisome proliferator activated receptor α and carnitine palmitoyltransferase, were associated with an insufficient energy supply within the BDL group post‑PH. Notably, the expression levels of fatty acid synthesis‑associated genes, including sterol‑regulatory element‑binding protein‑1c, acetyl‑coA carboxylase 1 and fatty acid synthase were also reduced within the BDL group, which accounted for the reduced triglyceride content and fatty acid utilization. Further investigation revealed that overactivated farnesoid X receptor (FXR) signaling may inhibit fatty acid synthesis within BDL group rats. Collectively, the role of triglycerides in liver regeneration following PH in extra‑cholestatic livers was identified in the present study. Additionally, the results indicated that overactivated FXR signaling‑induced triglyceride reduction is associated with insufficient energy supply and therefore contributes to the extent of impairment of liver regeneration following PH within extra‑cholestatic livers.

Clinical significance of expression of proliferating cell nuclear antigen and E-cadherin in gastric carcinoma.

AIMto investigate the expression of proliferating cell nuclear antigen (PCNA) and E-cadherin in gastric carcinoma and to analyze their clinical significance.METHODSA total of 146 patients were selected for this study, including 38 patients with intestinal metaplasia, 42 with dysplasia, and 66 with primary gastric cancer. In addition, 40 patients with normal gastric tissues were selected as controls. The expression of PCNA and E-cadherin was detected by immunohistochemistry. Differences in PCNA and the E-cadherin labeling indexes among normal gastric mucosa, intestinal metaplasia, dysplasia, and gastric carcinoma were compared. Subjects with normal gastric tissues were assigned to a normal group, while gastric cancer patients were assigned to a gastric cancer group. The difference in PCNA and E-cadherin expression between these two groups was compared. The relationship between expression of PCNA and E-cadherin and clinicopathological features was also explored in gastric cancer patients. Furthermore, prognosis-related factors, as well as the expression of PCNA and E-cadherin, were analyzed in patients with gastric cancer to determine the 3-year survival of these patients.RESULTSThe difference in PCNA and the E-cadherin labeling indexes among normal gastric mucosa, intestinal metaplasia, dysplasia, and gastric carcinoma was statistically significant (P < 0.05). During the transition of normal gastric mucosa to gastric cancer, the PCNA labeling index gradually increased, while the E-cadherin labeling index gradually decreased (P < 0.05). The PCNA labeling index was significantly higher and the E-cadherin labeling index was significantly lower in gastric cancer than in dysplasia (P < 0.05). The expression of PCNA was significantly higher in the gastric cancer group than in the normal group, but E-cadherin was weaker (P < 0.05). There was a negative correlation between the expression of PCNA and E-cadherin in gastric carcinoma (r = -0.741, P = 0.000). PCNA expression differed significantly between gastric cancer patients with and without lymph node metastasis and between patients at different T stages. E-cadherin expression also differed significantly between gastric cancer patients with and without lymph node metastasis (P < 0.05). High T stage and positive PCNA expression were risk factors for the prognosis of patients with gastric cancer (RR > 1), while the positive expression of E-cadherin was a protective factor (RR < 1). The sensitivity, specificity, and accuracy of PCNA positivity in predicting the 3-year survival of patients with gastric cancer were 93.33%, 38.89%, and 0.64, respectively; while these values for E-cadherin negativity were 80.0%, 41.67%, and 0.59, respectively. When PCNA positivity and E-cadherin negativity were combined, the sensitivity, specificity, and accuracy were 66.67%, 66.67%, and 0.67, respectively.CONCLUSIONCombined detection of PCNA and E-cadherin can improve the accuracy of assessing the prognosis of patients with gastric cancer.

Expression of Proliferating Cell Nuclear Antigen (PCNA) in breast carcinoma and its association with Ki67 and tumor grade in mastectomy specimens

Background: Breast cancer ranks first among cancer in urban population of India, and 2nd most common in the rural population. Proliferative activity acts as an adjunct and gives prognostic importance in breast cancer. However, the best method for measurement of proliferative rate is controversial. The aim was to evaluate the association between PCNA labeling index (PCNA-LI) and Ki67 labeling index (Ki67-LI) and their correlation with histological grade. Methods: Seventy cases of simple and modified Radical Mastectomy specimens with the diagnosis of carcinoma breast were included in the study. H&E stained sections were examined for assessment of histological grade. IHC scoring was done for the markers Ki-67 and PCNA. Correlation of PCNA with Ki67 and tumor grade was performed. Result: 18 specimens were grade 1 tumors, 36 cases were grade 2 and 16 were grade 3. 90.0% of tumors were positive for PCNA index and Ki-67 positivity was observed in 85.7% cases, median and mean values of PCNA and Ki67 were 52.0, 51.64+/-37.475, and 24.50, 26.17± 24.183 respectively. Final score was divided into low and high index using median value as a cut off. Ki-67 LI showed strong positive correlation with histological grade (p=0.0001). PCNA LI showed positive correlation with histological grade (p=0.041) but no correlation was observed with Ki-67 LI (p=0.232) Conclusion: PCNA marker cannot substitute Ki67 but may be used as an additional marker to assess proliferative activity along with other markers. It may be useful to categorize subgroups with different grades of proliferative activity due to its easy applicability and assessment techniques. DOI: 10.21276/apalm.2017.1191

Experimental study on portal vein ligation combined with in situ splitting on liver regeneration in rats

Objective To investigate the effect of portal vein ligation combined with in situ splitting on liver regeneration in rats. Methods Seventy-five healthy male Sprague-Dawley rats were selected and randomly assigned into sham operation group (S), portal vein ligation group (PVL) and portal vein ligation combined with in situ splitting group (ALPPS). On 1 d, 3 d, 7 d, 10 d, 14 d after operation, the hepatic regeneration rate (HRR) of right median lobe was calculated, the serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), IL-6, HGF, VEGF were detected. mRNA of IL-6, HGF, TNF-α, TGF-β was assayed by real-time PCR, and the hepatic proliferating cell nuclear antigen (PCNA) labeling index was evaluated by immunohistochemistry. Results Comparing with PVL group, the HRR of the right median lobe obviously increased on day 3, 7, 10 and 14 in ALPPS group (P<0.05), and ALT and AST level were increased on 1 d (P<0.05). On day 1 and 3, the content of serum IL-6, HGF and VEGF were all increased in ALPPS group [(70.7±14.6) pg/ml vs. (134.2±31.4) pg/ml; (0.70±0.04) ng/ml vs. (0.74±0.02) ng/ml; (82.1±12.6) pg/ml vs. (103.5±14.7) pg/ml], respectively (P<0.05). The mRNA expression of IL-6, HGF, TNF-α, TGF-β and the PCNA labeling index were also increased in ALPPS group in comparison with those in PVL group on day 1 and 3 (P<0.05). All the indexes in the two groups were all higher than those in the group S (P<0.05). Conclusions Portal vein ligation combined with in situ splitting could significantly enhance liver regeneration. The possible mechanisms were related to the inflammation reaction and stress response caused by in situ splitting and up-regulation of cytokines in the regenerating lobe after portal vein ligation combined with in situ splitting, especially IL-6, HGF and TNF-α. Key words: Liver partition; Portal vein ligation; Staged hepatectomy; Liver regeneration; Future liver volume

The effects of quercetin on liver regeneration after liver resection in rats.

The aim of the present study was to assess the influence of quercetine (QE) on liver regeneration after partial hepatectomy (PH) in rats. A total of 24 male Wistar albino rats were divided into three groups: sham-operated (SH), PH and PH+QE; each group contain 8 animals. The rats in QE-treated groups were given QE (15 mg/kg body weight) once a day i.p., for 7 days starting 3 days prior to hepatectomy operation. At 7 days after resection, liver samples were collected. The malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) levels were estimated in liver homogenates. Moreover, histopathological examination, mitotic index (MI), proliferating cell nuclear antigen labelling, proliferation index (PI), transferase-mediated dUTP nick end-labelling assay, apoptotic index (AI) were evaluated at 7 days after hepatectomy. As a result, QE significantly increased MI, PI, and significantly decreased AI in PH rats. Additionally, QE remarkably inhibited the elevation of MDA, restored impaired antioxidant SOD activity and GSH level, and also attenuated hepatic vacuolar degeneration and sinusoidal congestion. These results suggested that QE treatment had a beneficial effect on liver regenerative capacity of the remnant liver tissue after hepatectomy, probably due to its antioxidative, antiapoptotic and proliferative property.

The effects of quercetin on liver regeneration after liver resection in rats.

The aim of the present study was to assess the influence of quercetine (QE) on liver regeneration after partial hepatectomy (PH) in rats. A total of 24 male Wistar albino rats were divided into three groups: sham-operated (SH), PH and PH+QE; each group contain 8 animals. The rats in QE-treated groups were given QE (15 mg/kg body weight) once a day i.p., for 7 days starting 3 days prior to hepatectomy operation. At 7 days after resection, liver samples were collected. The malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) levels were estimated in liver homogenates. Moreover, histopathological examination, mitotic index (MI), proliferating cell nuclear antigen labelling, proliferation index (PI), transferase-mediated dUTP nick end-labelling assay, apoptotic index (AI) were evaluated at 7 days after hepatectomy. As a result, QE significantly increased MI, PI, and significantly decreased AI in PH rats. Additionally, QE remarkably inhibited the elevation of MDA, restored impaired antioxidant SOD activity and GSH level, and also attenuated hepatic vacuolar degeneration and sinusoidal congestion. These results suggested that QE treatment had a beneficial effect on liver regenerative capacity of the remnant liver tissue after hepatectomy, probably due to its antioxidative, antiapoptotic and proliferative property.

THE EFFECTIVE ROLE OF GINGER AGAINST THE OVOTOXICITY INDUCED BY 7, 12-DIMETHYLBENZ[A]ANTHRACENE IN RAT: ULTRASTRUCTURAL AND IMMUNOHISTOCHEMICAL STUDIES

The present study aims to evaluate the role of ginger extract against the ovotoxic and/or carcinogenic effects induced by 7, 12-dimethylbenz[a]anthracene (DMBA) in the ovarian tissues of female albino rats. Ginger (Zingiber officinale) has acquired considerable attention as a botanical dietary due to its active biological constituents of phenolic substances ([6]-gingerol and [6]-paradol) that possess anti-inflammatory, anti-oxidative properties and exerts substantial anti-carcinogenic activities. In the present study, a single dose of 40 mg/kg of 7,12-dimethylbenz[a]anthracene was used to induce ovotoxicity, and aqueous extract of ginger concentration (120 mg/ml) every other day as a possible protective substance. The ultrastructural observations of DMBA-treated group showed some differences such as fibroblast cells with condensed chromatin, primordial follicles area exhibiting scattered cytoplasmic organelles and ruptured-plasma membrane, some granulosa of the secondary follicles showing cytoplasmic vacuolization and some others revealing multi-nucleated. The ultrastructural observations of combined DMBA and ginger treated rats revealed more or less normal fibroblasts, primordial follicular cells, granulosa of secondary follicles and improvement in the number and shape of mitochondria. Concerning the immunohistochemical observations, the ovary of DMBA treated rats showed a significant increase in the proliferating cell nuclear antigen labeling index (PCNA LI, P ≤ 0.001) with respect to control and/or ginger treated rats. However, DMBA and ginger treated rats showed a significant increase in the PCNA LI (P ≤ 0.01) with respect to control and/or ginger treated rats and a significant decrease (P≤ 0.001) with those of DMBA treated rats. In conclusion, ginger may play a possible role as a protective agent against the ovotoxicity and/or carcinogenicity of DMBA showing an improvement of the ovarian tissue.

Synergistic Inhibitory Effect of Rosuvastatin and Angiotensin II Type 2 Receptor Agonist on Vascular Remodeling.

We investigated the possibility that coadministration of rosuvastatin and compound 21 (C21), a selective angiotensin II type 2 (AT2) receptor agonist, could exert synergistic preventive effects on vascular injury. Vascular injury was induced by polyethylene cuff placement on the femoral artery in 9-week-old male C57BL/6J mice. Mice were treated with rosuvastatin and/or with C21 after cuff placement. Neointima formation was determined 14 days after the operation and cell proliferation, and superoxide anion production and expression of inflammatory cytokines were examined 7 days after cuff placement. Neointima formation was significantly attenuated by the treatment of rosuvastatin (5 mg kg(-1) day(-1)) or C21 (10 μg kg(-1) day(-1)), associated with the decreases in proliferating cell nuclear antigen (PCNA) labeling index, oxidative stress, and the expression of inflammatory markers. Treatment with a noneffective dose of rosuvastatin (0.5 mg kg(-1) day(-1)) plus a low dose of C21 (1 μg kg(-1) day(-1)) inhibited the PCNA labeling index, superoxide anion production, mRNA expressions of NAD(P)H subunits, and mRNA and protein expressions of inflammatory markers associated with marked inhibition of neointima formation. Angiotensin II type 1 (AT1) receptor mRNA expression did not differ the groups. By contrast, AT2 receptor mRNA expression was increased by administration of C21 at the dose of 10 μg kg(-1) day(-1) but not by C21 at the dose of 1 μg kg(-1) day(-1) or rosuvastatin. The combination of rosuvastatin and AT2 receptor agonist exerted synergistic preventive effects on vascular remodeling associated with the decreases in cell proliferation, oxidative stress, and inflammatory reaction. That could be a powerful approach to vascular disease prevention.

Hyperbaric oxygen therapy stimulates colonic stem cells and induces mucosal healing in patients with refractory ulcerative colitis: a prospective case series

BackgroundHyperbaric oxygen (HBO) is used as part of treatment in a variety of clinical conditions. Its use in the treatment of ulcerative colitis has been reported in few clinical reports.ObjectiveWe...

Autologous adipose tissue‑derived mesenchymal stem cells are involved in rat liver regeneration following repeat partial hepatectomy.

Adipose tissue-derived mesenchymal stem cells (ADSCs) have been considered to be attractive and readily available adult mesenchymal stem cells, and they are becoming increasingly popular for use in regenerative cell therapy, as they are readily accessible through minimally invasive techniques. The present study investigated whether autologous ADSC transplantation promoted liver regeneration following a repeat partial hepatectomy in rats. The rats were divided into three groups as follows: 70% partial hepatectomy (PH) group; repeat PH (R-PH) group and R-PH/ADSC group, subjected to R-PH and treated with autologous ADSCs via portal vein injection. In each group, the rats were sacrificed at different time points postoperatively in order to evaluate the changes in liver function and to estimate the liver regenerative response. The expression of proliferating cell nuclear antigen (PCNA) labeling index in the liver was measured using immunohistochemistry. The expression levels of hepatocyte growth factor (HGF) mRNA were measured using reverse transcription polymerase chain reaction. The results showed that regeneration of the remaining liver following R-PH was significantly promoted by ADSC transplantation, as shown by a significant increase in liver to body weight ratio and the PCNA labeling index at 24 h post-hepatectomy. Additionally, ADSC transplantation markedly inhibited the elevation of serum levels of alanine aminotransferase, aspartate aminotransferase and total bilirubin, increased HGF content and also attenuated hepatic vacuolar degeneration 24 h postoperatively. Furthermore, the liver was found to almost fully recover from hepatocellular damage due to hepatectomy among the three groups at 168 h postoperatively. These results indicated that autologous ADSC transplantation enhanced the regenerative capacity of the remnant liver tissues in the early phase following R-PH.