【Abstract】ObjectiveTo investigate the effects of liver transplantation on splenic function in rats with hepatic cirrhosis. MethodsHepatic cirrhosis model was established in rats by subcutaneous injections of carbon tetrachloride. Liver transplantation model was established with twocuff technique. Spleen index, morphological changes of spleen were observed before and after liver transplantation in hepatic cirrhosis rats. Spleen T lymphocyte subgroups before and after liver transplantation were also assayed by immunofluorescence staining and flow cytometry. ResultsBefore liver transplantation, spleen index was increased from (2.42±0.11) mg/g to (3.62±0.14) mg/g， P＜0.01; pathological examination of spleen samples showed that the areas of white pulp were decreased from (23.47±2.30)% to (7.70±2.01)%， P＜0.01， and the areas of spleen trabecula were increased from (1.75±0.61)% to (4.46±0.71)%， P＜0.01. Meanwhile, the ratio of CD4/CD8 of spleen T lymphocyte subgroups was decreased from 2.67±0.15 to 1.18±0.15, P＜0.01. After liver transplantation, spleen index was decreased from (3.62±0.14) mg/g to (2.62±0.11) mg/g， P＜0.01; pathological examination of spleen showed that the areas of white pulp were increased from (7.70±2.01)% to (15.07±1.97)%， P＜0.01, and those of spleen trabecula were decreased from (4.46±0.71)% to (3.11±0.51)%， P＜0.05. Meanwhile, the ratio of CD4/CD8 of spleen T lymphocyte subgroups was increased from 1.18±0.15 to 2.32±0.11, P＜0.01. ConclusionImpaired function of spleen resulting from liver function damage can be improved in rats with hepatic cirrhosis after liver transplantation.
ObjectiveTo explore the effect of exogenous ghrelin on early recovery of rats after subtotal gastrectomy. MethodsTwelve rats undergoing subtotal gastrectomy (B-Ⅰtype) were randomly divided into two groups, and saline or ghrelin was intraperitoneally injected in two groups, respectively. The body weight and daily food intake were measured before operation and on 1-7 d after operation. Rats were killed on day 7 after operation and the expressions of ghrelin mRNA in the fundus of stomach and anastomotic stoma was determined by realtime fluorescent quantitative PCR assay. The anastomotic bursting pressure and hydroxyproline content of anastomotic stoma tissues were also detected. ResultsThere was no significant difference (P＞0.05) in pre and postoperative body weight between two groups. Gradual decrease in postoperative body weight among the rats of saline group was observed which was significantly lower than that before operation (Plt;0.01). Body weight reached it’s lowest on day 1 after operation (Plt;0.01), after which it gradually increased but was still lower than that before operation (Plt;0.01). The postoperative body weight of rats in ghrelin group gradually decreased too, and was also significantly lower than preoperative body weight (Plt;0.01), except for the day 1 after operation (P=0.693). It reached the lowest on day 4 after operation (Plt;0.01), then it gradually increased but was still lower than that before operation (Plt;0.05 or Plt;0.01). The cumulative food intake of rats in ghrelin group was （52.50±6.77） g, which was significantly higher than that in saline group 〔(45.67±7.47) g〕, Plt;0.05. On day 7 after operation, relative expression of ghrelin mRNA in the fundus of stomach of rats in ghrelin group was 0.08±0.04, which was significantly lower than that in saline group (0.22±0.07), Plt;0.01. Compared with saline group, ghrelin-treated rats displayed significantly higher bursting pressure 〔（155.83±6.62） mm Hg vs. （172.33±10.44） mm Hg, Plt;0.05〕 higher hydroxyproline content 〔 (0.43±0.05) μg/mg wet tissue vs. （0.50±0.29） μg/mg wet tissue, Plt;0.01〕 at the anastomotic stoma. ConclusionGhrelin may effectively promote the early recovery of rats after subtotal gastrectomy.
Objective To investigate the effects of granulocyto-colony stimulating factor (G-CSF) on the mobil ization of endothel ial progenitor cells (EPCs) in the rats with myocardial infarction (MI), to observe the density of neovascularization and the mRNA expressions of vascular endothel ial growth factor (VEGF) and its receptor (Flk-1) in the border area of MI. Methods Thirty-six adult male rats (weighing 250-280 g) were divided randomly into control group, MI group, and G-CSF group. In MI group and G-CSF group, the models of MI were establ ished by left anterior descenting coronary artery l igation and were treated with intraperitoneal injection of sal ine (0.3 mL/d) or G-CSF [30 μg/(kg•d)] for 5 days. In control group, after open chest operation, chest was closed without treatment. The level of EPCs was surveyed and the plasma concentrations of VEGF and C-reaction protein (CRP) were measured at 7 days. The mRNA expressions of VEGFand its receptor Flk-1 in the border area of infarct myocardium were determined through RT-PCR. Results Compared withcontrol group, the number of circulating white blood cell (WBC) and EPCs levels, and the serum concentrations of VEGF and CRP were all significantly increased in MI group and G-CSF group (P lt; 0.05); when compared with MI group, the number of circulating WBC and EPCs levels, and the serum concentrations of VEGF were increased and the concentration of CRP was decreased in G-CSF group (P lt; 0.05). Compared with control group, the mRNA expressions of VEGF and Flk-1, and the density of neovascularization in the border area of infarct myocardium were increased in MI group and G-CSF group, whereas those in G-CSF group were significantly augmented compared with MI group (P lt; 0.05). Conclusion In the rats with MI, G-CSF could promote EPCs mobil ization, increase the mRNA expressions of VEGF and Flk-1, and augment the density of neovascularization in the border area of infarct myocardium.
Objective To explore the effects of several immunosuppressants on the cell numbers of cultured rat macrophages and Schwann’s cells. Methods The macrophages and Schwann’s cells were cultured from the newborn Wistar rats. Different concentrations of methylprednisolone(10-3, 10－4,10－6 and 10－8 mol/L), CsA(10－5, 10－6, 10－7 and 10－8 mol/L) and FK506(10-6, 10－7, 10－8 and 10－9 mol/L) were administrated to the cells, while control group was given no drugs. Twentyfour, 48 and 72 hours after administration, the cells from different concentrations were measured with MTT methods respectively. Theresults were compared and analyzed statistically. Results Only high concentration methylprednisolone (10-4 mol/L) and a certain range of concentrations of CsA (10-6，10-7 and 10-8 mol/L) and FK506 (10-7,10-8 and 10-9 mol/L) can provide protection to culturedrat macrophages. Under most concentrations, CsA and FK506 had no effects onthe cell number of cultured rat Schwann’s cell. Only with high concentration CsA (10-5 mol/L) and methylprednisolone (10-3 mol/L) could significantly decreased the cell number of Schwann’s cell. Long time (72 hours) and low dosage (10-8 mol/L) administration of methylprednisolone could significantlyprotect Schwann’s cell. Conclusion High concentration methylprednisolone and some certain concentration CsA and FK506 can protect cultured rat macrophages. But high concentration CsA and methylprednisolone prohibit the proliferation of Schwann’s cells. Only long time and low dosage methylprednisolonecan protect cultured rat Schwann’s cells.
ObjectiveThe optimal target of deep brain stimulation (DBS) for treating intractable epilepsy is still undefined. Cumulative studies suggest that the mediodorsal thalamic nucleus (MD) is involved in seizure activity, the purpose of this study was to investigate the effect of high frequency stimulation in MD on pentylenetetrazole (PTZ)-induced seizures in rats. MethodsThe experimental rats (Male Sprague-Dawley rats 280-350 g) were all provided by Experimental Animal Center, Zhejiang Academy of Medical Science, Hangzhou, China. The rats were given unilateral or bilateral stimulation of the MD at 100 Hz (HFS group) and sham stimulation, others were given unilateral stimulation of the MD at 1 Hz (LFS group). EEGs in the cortex and seizure behavior were recorded with the Neuroscan system at the same time. ResultsNeither LFS nor HFS of the MD changed the latency to the first spikes or EEG manifestations for stage 3 and stage 5 seizures; animals receiving unilateral or bilateral HFS of the MD decreased the number of stage 5 EEG seizure synchronized with the convulsive episodes; LFS and sham stimulation showed multiple periods of continuous spikes which accompanied stage 5 or stage 4 seizures. HFS of unilateral or bilateral MD, but not LFS, decreased the seizure stage, the number of clonic movement episodes, and the duration of acute PTZ-induced seizures. The average latency to onset of myoclonic jerks did not differ among groups. Unilateral and bilateral HFS of the MD had a similar antiepileptic effect. ConclusionHFS of the MD may be of value as a new antiepileptic approach for patients with generalized epilepsy, besides, the seizure model, should be fully considered in clinical application.
ObjectiveTo assess the methods, processes and evaluation criteria of burn models in rats. MethodsDatabases including MEDLINE, EMbase, CBM, CNKI, VIP and WanFang Data were electronically searched from inception to February 2016. The animal studies of burn models development in rats were included. Two reviewers independently screened literatures, extracted data, assessed the quality of included studies, and analyzed the outcomes. ResultsFifteen studies including 38 burn groups were identified. The results indicated:(1) depth of burn:50.00% burn models were partial thickness burn (II°), and 21.05% were full thickness burn (III°). (2) methods of induction:80.00% burn models were developed through high temperature liquid or solid tools. (3) burn sites:93.33% burns were on the back of rats. (4) induction temperature:66.67% induction temperature was between 80℃ and 100℃. (5) induction time:86.67% induction time was between 3 seconds and 100 seconds. (6) anesthesia:40.00% rats were anesthetized with intraperitoneal injection of pentobarbital sodium. (7) skin preparation:60.00% studies reported the skin was prepared by physical and chemical methods. (8) housing post surgery:13.33% studies reported the housing temperature post surgery. (9) intervention post surgery:13.33% studies reported antishock treatment. (10) assessment criteria:almost all studies evaluated the depth of burn through the macroscopic and microcosmic assessment. ConclusionBased on current animal studies, most burn models in rats are partial thickness and full thickness burn on the back of rats. Burn is inducted mainly by 80℃ to 100℃ hot liquid or solid tools within 30 seconds, according with skin preparation, anesthesia, antishock or analgesia management. The depth of burn is evaluated by the macroscopic and microcosmic assessment. However, there is no standard for the methods, processes, assessment and reporting of development of burn model in rats.
ObjectiveTo explore the mechanism of lung injury in Sprague-Dawley (SD) rats induced by acute organic phosphorus pesticides (AOPP) by observing the changes of the blood serum nuclear factor (NF)-κB consistence, NF-κB level of lung tissue and lung coefficient. MethodNinety-six healthy male SD rats (six weeks old) were randomly divided into group A (control, n=48) and group B (poison, n=48). The rats of group B were given omethoate by gavage (45 mg/kg), and the rats of group A accepted normal saline. Then the rats were killed at designated observing points (30 minutes; 3, 6, 12, 24, and 48 hours), and the lung coefficient, blood serum NF-κB consistence and NF-κB level of lung tissue were measured. At the same time, we observed the pathological changes of the rats' lung tissue. ResultsCompared with group A, blood serum NF-κB consistence, NF-κB level of lung tissue and the level of lung coefficient in group B were significantly higher (P<0.01). The lung tissues of group A were normal at each time point, but in group B, the lung pathological changes gradually appeared 30 minutes later with pulmonary interstitial engorging, alveolar septum widening and some alveolus being full of red blood cells, and this situation reached its peak at hour 12. Then it gradually mitigated from 24 to 48 hours. ConclusionThere are significant increases in blood serum NF-κB consistence and NF-κB level in lung tissues in rats with lung injury induced by omethoate poisoning. The NF-κB may play a role in the process of lung injury induced by organophosphorus pesticide.
Objective To study the relationship between the expression ratio of induced nitric oxide synthase (iNOS) over glial fibrillary acidic protein (GFAP) and the time of injury after brain concussion in rat, in order to acquire a new visual angle for determining injury time of cerebral concussion. Methods Eighty-five healthy Sprague-Dawley rats were divided into three groups randomly: model group (n=25), experimental group (n=55), and control group (n=5). The rats in the model group were used to confirm the attack hight to make the model of brain concussion; according to the time of execution, rats in the experimental group were then subdivided into 11 groups with 5 rats in each subgroup, and their execution time was respectively hour 0.5, 1, 3, 6, 12, 24, 48, 96, 168, 240, and 336; the rats in the control group were executed after fed for 24 hours. After the model of cerebral concussion was established through freefalling dart method, hematoxylin-eosin staining and immunohistochemistry staining of iNOS and GFAP were conducted for the brain of the rats. All related experimental results were studied by using microscope with image analytical system and homologous statistics. Results The ratio of positive expression of iNOS over that of GFAP increased gradually during hour 0.5- 3 after injury in brain (from 5.03 to 10.47). At the same time, the positive expression of iNOS increased significantly (from 14.61% to 37.45%). However, the increase of the positive expression of GFAP was not obvious. Between hour 3 and 12, the ratio began to decline to 4.98, which was still at a high level, and during the same time period, the positive expressions of iNOS and GFAP also experienced the same change pattern. Later, the ratio began to decline between hour 12 and 336 after injury (from 4.98 to 0.95). All ratios at this time were lower than those between hour 0.5 and 12. The positive expression of iNOS and GFAP both increased to a climax before declining. Conclusions The ratio of positive expression of iNOS over GFAP and the respective change pattern of iNOS and GFAP can be used as the evidence of estimating the injury time of cerebral concussion. We can use the ratio of two or more markers to provide a new visual angle for concluding the concussion injury time.
ObjectiveTo investigate the effect of post-conditioning with fospropofol disodium on hepatic ischemiareperfusion (I/R) and its possible mechanism in rats. MethodsForty-eight Sprague-Dawley rats were randomly divided into four groups, including sham group (S), control group (C), propofol group (P) and fospropofol disodium group (F). According to the different periods after reperfusion, each group was further divided into 2-hour and 4-hour reperfusion subgroups respectively (n=6 in each subgroup), named S2h, C2h, P2h, and F2h subgroups and S4h, C4h, P4h, and F4h subgroups. The livers of rats were reperfused after hepatic ischemia for one hour. In the beginning of reperfusion, normal saline was infused intravenously in group S and group C continuously, propofol was infused intravenously in group P continuously, fospropofol disodium was infused continuously in group F. The blood was sampled at the end of ischemia and reperfusion for assay of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The bcl-2 and bax protein contents in liver tissue were detected by immunohistochemical analysis, and liver samples were stained with hematoxylin-eosine for histological observation and damage degree evaluation by counting the proportion of necrosis cells. ResultsThe activity of ALT and AST, the rate of necrosis cells and the amount of bcl-2 and bax protein after reperfusion in group C, group P and group F were higher than those in group S at matched reperfusion time points (P<0.05). The activity of ALT and AST, the proportion of necrosis cells and bax protein contents decreased in group P and group F, compared with group C at the same reperfusion time points, while the contents of bcl-2 protein were significantly increased (P<0.05). ConclusionFospropofol disodium can alleviate hepatic injury induced by ischemia-reperfusion in rats, in which the bcl-2 and bax protein may play important roles.