Expression of IL-6 mRNA in endometrium with endometriosis

AIM: The purpose of the present study was to explore the relationship between interleukin-6 mRNA expression and endometriosis. METHODS: Using the rat model, IL-6 mRNA expression in the endometrium was examined by RT-PCR. RESULTS: The expression of IL-6 mRNA in control rats did not change at 2, 4, 6 and 8 weeks after sham operation (P>0.05), but in model rats it gradually increased at 2, 4, 6 and 8 weeks after endometriosis (P<0.01). The expression of IL-6 mRNA in uterine endometrium with endometriosis was lower than in endometriotic tissue, but higher than in endometrium from healthy controls. CONCLUSION: The IL-6 mRNA expression may contribute to the development of endometriosis . The increase in IL-6 mRNA expression may promote the implantation and growth of endometriotic tissue.     

Time-related effects of danazol treatment in experimentally induced endometriosis in the rat

AIM and METHODS: The purpose of the study was to characterize the time-related effect of Danazol therapy on endometriosis explant using the rat model. Endometriosis was induced in mature female rats. One group of treated animals as well as controls were sacrificed at 2,4,6 and 8 weeks after treatment at which time the explant was evaluated. RESULTS:Explant volume was significantly reduced in all treatment groups, and the effect was more significant in animals treated for 4 weeks than those treated for only 2 weeks. CONCLUSION: Danazol treatment can cause gradual regression of endometrial explant in a time-related manner.     

Effects of swimming on experimental endometriosis in rats

AIM To evaluate the effect of swimming on experimental endometriosis in rats. METHODS 80 female SD rats were divided into 8 groups, including control group, model group and animals performed light exercise （swimming once a week）, moderate exercise （swimming 3 times a week）, and intense exercise （swimming 5 times a week） before or after endometriosis induction,10 rats in each group. The mRNA and protein expressions of fatty acid synthase （FAS）, matrix metalloproteinase 9 （MMP9） and proliferating cell nuclear antigen （PCNA） in endometrium of rats were detected. RESULTS The swimming before the induction of the edometriosis lesions did not prove to have aprophylactic role against endometriosis, whereas the swimming after induction of the lesions had a beneficial effect regardless of frequency, with a greater reduction in the groups practicing moderate and intense activity （P<0.05）, an increase in FAS levels and a decrease in MMP9 and PCNA levels were also observed （P<0.05）. CONCLUSION Swimming after induction of the edometriosis is beneficial for the treatment of endometriosis, the mechanism may be related to the expression of FAS, MMP9 and PCNA protein.     

Effects of (+)-2-(1-hydroxyl-4-oxocyclohexyl)ethyl caffeate at different doses on rat arthritis model and whole blood inflammation model

AIM:To evaluate the pharmacodynamics of (+)-2-(1-hydroxyl-4-oxocyclohexyl)ethyl caffeate (HOEC), and to explore the possible causes of non-dose-dependent effects of HOEC on collagen-induced arthritis (CIA) rats using the arachidonic acid (AA) metabolic model in rat whole blood. METHODS:The rat CIA model was used to study the treatment with HOEC at 3 doses. The expression of cytoplasmic phospholipase A₂ (cPLA₂), 5-lipoxygenase (5-LOX) and cyclo-oxygenase-2 (COX-2) was assayed by immunohistochemical method. The effects of HOEC and its in vivo metabolite caffeic acid (CA) on AA metabolite in rat whole blood were measured by ELISA. RESULTS:HOEC had a therapeutic effect on rat CIA, but the curative effect at low dose and middle dose (1 and 3 mg/kg) was better than that at high dose (10 mg/kg). The expression levels of cPLA₂, 5-LOX and COX-2 in joint tissues were decreased. HOEC inhibited the metabolites of LOX and COX pathways in the rat whole blood AA metabolic model, while the inhibitory effect of CA on these metabolites was weaker than that of HOEC. CONCLUSION:The anti-inflammatory effect of HOEC on rat CIA may be associated with the inhibition of cPLA₂, 5-LOX and COX-2 expression in the joint tissues. The non-dose-dependent therapeutic effect of HOEC on rat CIA may due to the weaker inhibitory activity of CA on AA metabolic model in rat whole blood than that of HOEC.     

Effects of N-acetylcysteine on Clara cells and CC16 in rat COPD model

AIM: To investigate the effects of N-acetylcysteine (NAC) on the number of Clara cells and secretion of Clara cells secretory protein (CC16) in rat chrohic obstructive pulmonary disesae (COPD) model.METHODS: Thirty rats were divided into control, COPD and NAC groups (n=10). The change of Clara cell ultrastructure was detected through transmission electron microscope. The number of Clara cells and synthesis of CC16 were measured by immunohistochemistry. The CC16 levels in bronchoalveolar lavage fluid (BALF) and serum were tested by ELISA. The level of CC16 mRNA in lung was determined by RT-PCR.RESULTS: The percentage of Clara cells in terminal bronchioles in the COPD group was significantly decreased than that in the control (P<0.01), and the percentage in NAC group was significantly higher than that in COPD group (P<0.01). The levels of CC16 in the BALF and serum in COPD group were significantly lower than those in control group (P<0.01, respectively), and the levels of CC16 in NAC group were significantly higher than those in COPD group (P<0.05, respectively). The expression of CC16 mRNA in COPD group was weaker than that in control group and NAC group (P<0.01, respectively).CONCLUSION: The number of Clara cells and the secretion of CC16 decrease in a rat model of COPD. Antioxidant NAC can enhance the synthesis and secretion of CC16, which may be a mechanism for the suppression of airway inflammation.     

Effects of mifepristone on ultrastructure of human endometrium in the early secretory phase

AIM:To investigate the influence of mifepristone on ultrastucture of human endometrium in the early secretory phase. METHODS: Endometrial tissue was obstained from 10 patients of reproductive age, who underwent a hysterectomy within 1 week postovulatory for gynecologic diseases not involving the endometrium. Patients were divided into mifepristone group (n=5) and control group (n=5) randomly. Each patient in the mifepristone group had taken 25 mg mifepristone per os 24 h before the operation was performed, while none of the control group had taken mifepristone. After removal of uterus, endometrial tissue was immediately acquired and prepared for electron microscopic examination. RESULTS:In comparison with the control group, the endometrial tissue in mifepristone group displayed the following distinctly morphological changes: (1) In the endometrial epithelium neither nucleolar channel system nor giant mitochondrium was seen, and subnuclear glycogen accumulation was seldom observed, but giant lysosomes were frequently found. (2) The intercellular spaces of the epithelium were narrow and straight, the indigitations of lateral plasma membranes were rarely visible. (3) Cytolysis and karyopyknosis of stroma cells and extravasal red cells were repeatedly observed.CONCLUSION:The above ment ioned morphological changes in endometrium in the early secretory phase caused by mifepristone are undoubtedly sufficient to prevent implantation.Consequently, mifepristone may have a contraception effect.     

A rat model of repeated seizures induced by pentylenetetrazol at different maturational stages and the role of NF-κB in the pathogenesis of epilepsy

AIM: To observe histopathologic changes and NF-κB expression in hippocampus in neonatal and matural rats after repeated seizures, and to explore the role of NF-κB in the pathogenesis of epilepsy in premature brain of rats. METHODS: Neonatal rats and mature rats were divided into 2 experimental groups at 10 days and 60 days after birth (P10 and P60). Convulsions were induced by repeated injection of pentylenetetrazol (PTZ) intraperitoneally for first 5 days. The animals in control group were injected with NS at the same volume in the same conditions. The neurons in CA1, CA3, dentate granule (DG), as well as in hilar were counted by thionin staining, in order to observe the profile of the necrosis and apoptosis. NF-κB expression was examined by immunohistochemistry assay. Timm's method of silver sulfide staining was adopted to observe the mossy fiber sprouting. RESULTS: (1) In immature rats (10 days old), neurons in CA1, CA3 and hilar demonstrated no differences from controls, whereas adult rats (P60) had a significant decrease in number of neurons in CA1 and CA3 (8.22±1.88, 5.62±1.68 vs 6.31±1.50, 3.62±1.40). In adult rats, neurons in dentate granule showed no differences with controls, whereas immature rats with daily seizures had a significant increase (23.25±3.06 vs 16.25±1.58). (2) There was prominent sprouting in the CA3 stratum pyramidal layer in all experimental rats after 5 daily seizures, regardless of the age. However, the degree of sprouting was significantly different between the two experimental groups (3.25±1.03 vs 1.50±0.92, P<0.05). (3) NF-κB was highly expressed in CA3, CA1 and DG after 24 hours by PTZ-kindling, whereas it was little expressed in control group. NF-κB expression was higher in P10 experimental rats than that in P60 rats. CONCLUSION: No cell loss was observed in hippocampus in neonatal rats after recurrent kindling. The high expression of NF-κB may be one of the important molecular mechanisms underlying the special resistance of the neurons in premature brain to the epileptic cerebral lesions.     

p38MAPK expression and apoptosis in the rat unilateral ureteral obstruction model

AIM: To investigate the expression of p38 mitogen-activated protein kinase (p38MAPK) in the kidney after unilateral ureteral obstruction (UUO) in rats and the functional role of it on apoptosis and fibrosis.METHODS: Eighteen Wistar rats underwent UUO were killed at 3, 7, 14 days. Additional 7 rats were sham operated. Histological changes were observed by HE and Masson staining. Immunohistochemistry study was performed on renal tissue for proliferating cell nuclear antigen (PCNA). Apoptotic cells were determined by terminal deoxynucleotidyl transferase- mediated dUTP nick end labeling (TUNEL) and the electrophoresis analysis of genomic DNA. Western blotting of cysteinyl aspartate specific proteinase-3 (caspase-3), p38MAPK and p-p38MAPK were measured.RESULTS: UUO induced a significant increase in renal tubular and interstitial cell apoptosis, immunohistochemistry of PCNA and Western blotting of caspase-3, p-p38MAPK as well as severe morphology changes. However, there was no significant difference between UUO and the control in Western blotting of p38MAPK.CONCLUSION: An in vivo model of renal fibrosis after UUO demonstrates that activated or phosphorylated p38MAPK plays a role in apoptosis of renal tubulointerstitial cells.     

Effect of pidotimod on renal function and inflammatory response in rat IgA nephropathy model

AIM:To explore the effect of pidotimod on the renal function in IgA nephropathy (IgAN) rat model, and to further study whether this effect is related to the inhibition of inflammatory response. METHODS:The SD rats (n=36) were randomly divided into control group, IgAN model group, IgAN with prednisone treatment group and IgAN with pidotimod treatment group, with 9 rats in each group. The IgAN model was induced by consecutive oral administration of bovine gamma globulin (BGG) for 8 weeks followed by injection of BGG through tail vein for 3 d. After the IgAN model was established, the drug was continuously used for 4 weeks. At the end of the treatment, the urine protein, serum creatinine and blood urea nitrogen were examined by an automated analyzer. IgA deposition in the renal tissues was observed by immunofluorescence staining. The mRNA expression levels of renal fibrosis markers transforming growth factor-β1 (TGF-β1) and fibronectin 1 in the renal tissues were detected by RT-qPCR. The mRNA and protein levels of pro-inflammatory cytokines interleukin-1β (IL-1β) and IL-6 in the renal tissues were determined by RT-qPCR and Western blot, respectively. RESULTS:No significant difference of the body weight was observed in different groups. Compared with control group, the content of urine protein, serum creatinine and blood urea nitrogen were significantly increased (P<0.01), whereas those were reversed by pidotimod treatment. The results of immunofluorescence staining showed that pidotimod inhibited IgA deposition in the IgAN rats. Pitomod treatment inhibited the mRNA expression levels of renal fibrosis markers TGF-β1 and fibronectin 1, and the mRNA and protein levels of pro-inflammatory cytokines IL-1β and IL-6 in the renal tissues of IgAN rats. CONCLUSION:Pidotimod alleviates IgAN progression in rats by inhibition of inflammatory response.     

Effects of WJMSC transplantation on cesarean scar defect in a rat model

AIM: To evaluate the effects of transplantation of Wharton jelly-derived mesenchymal stem cells (WJMSCs) on SD rats with previous cesarean scar defect (PCSD). METHODS: SD rats (n=50) were randomly divided into 3 groups:10 in normal operation group (N-O group), 20 in model control group (M-C group) and 20 in model treatment group (M-T group). On the 19th day of pregnancy, thread was used for continuous suture after the caesarean section (C-S) in N-O group. After C-S, LPS (1 g/L, 1 mL/kg) was injected in uterine muscle walls and electrocoagulation (power=10 W) was used in bilateral uterine incision with interrupted suture in both M-C and M-T groups. The rats in M-T group was further divided into A and B subgroups, with 10 rats each. The rats in group A were treated by intravenous injection and vaginal perfusion, both with 0.5 mL of 1×10⁹/L WJMSCs, while the rats in group B were treated by vaginal perfusion with 1 mL of WJMSCs, once a week, totally 3 times. The rats in M-C group were also divided into A and B subgroups (10 rats per group) and treated by the same way as M-T group, only except that saline was used instead of WJMSCs. At the 1st and 4th weeks after the treatment was finished, 6 and 4 rats were executed in every group, respectively. The uterine specimens with Masson and immunohistochemical staining were observed for determining the expression of actin and cytokeratin to reveal the repair of endometrial epithelium. RESULTS: The thickness of uterine muscle and volume density of actin in M-C group were significantly lower than those in N-O group, but the muscle collagen volume density and defect rate of endometrium were higher than those in N-O group. At the 1st and 4th weeks after treatment with WJMSCs, the muscular thickness and muscular actin concentration in M-T group (A and B) were significantly higher than those in M-C group, while the number of muscular collagen fibers were significantly lower than that in M-C group. The endometrial insufficiency in M-C group was higher than that in the other groups (P<0.05). CONCLUSION: WJMSCs promote the repair of the structure and function of myometrium in PCSD rats. No obvious difference between the effects of intravenous injection combined with vaginal perfusion, and vaginal perfusion alone on the repairing is observed.     

Establishment and evaluation of rat heart ischemia-reperfusion injury model in vivo

AIM: To establish and evaluate a rat model of heart ischemia-reperfusion injury in vivo. METHODS: Seventy-two male Sprague-Dawley rats weighing(250±50)g were randomly divided into sham operation group(sham), ischemia-reperfusion group(I/R) and normal group. The animals were anesthetized and heparinized. Myocardial ischemia-reperfusion was induced by ligating the left anterior descending coronary artery with "U-shape tube" for 35 min followed by 120 min or 240 min reperfusion in vivo. The heart infarct size was measured by triphenyltetrazolium chloride(TTC) staining. The myocardial cell apoptotic index was determined by the method of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling(TUNEL). Immunohistochemical method was used to detect the expression of Bcl-2 and Bax in rat ischemia myocardium. The blood level of MB isoenzyme of creatine kinase(CK-MB),cardiac troponin I(cTnI),nitric oxide(NO),malondialdehyde(MDA), total superoxide dismutase(T-SOD)and glutathione peroxidase(GSH-Px) were detected after reperfusion for 2 h and 4 h. RESULTS: Compared with normal group and sham group, there were obvious changes of ST-T segment and Q wave in the electrocardiogram of I/R group. The blood level of CK-MB, cTnI, NO, MDA and GSH-Px in I/R group increased(P<0.05,P<0.01) after reperfusion for 2 h and 4 h, and the blood level of T-SOD in I/R group after reperfusion for 2 h and 4 h also increased(P<0.05). The heart infarct size in I/R group was the largest as compared to other groups. Among these groups, the apoptotic index of I/R group was the highest and the Bcl-2/Bax ratio in I/R group decreased(P<0.01).CONCLUSION: The rat model of heart ischemia-reperfusion injury in vivo can be successfully established with the "U-shape tube". There are obviously changes of heart infarct size, blood level of CK-MB, cTnI, NO, MDA, T-SOD and GSH-Px, myocardial apoptotic index and Bcl-2/Bax ratio between I/R rats and control animals.     

Expression of VEGF and bFGF in rat model of pressure ulcer

AIM:To study the effects of vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) on the molecular pathogenesis of pressure ulcer.METHODS:SD rats were randomly divided into control group and experiment group. The pressure ulcer model was established by magnetic disk circulating compression method. HE staining was used to observe the pathological changes of the skin in the rats. The expression of VEGF and bFGF in the tissues was detected by immunohistochemical method. RESULTS:The expression of VEGF and bFGF in the tissues of rat Ⅲ-degree pressure ulcer was lower than that in the surrounding tissues and normal skin(P<0.01). The changes of VEGF and bFGF were consistent(κ=0.58). CONCLUSION:The expression levels of VEGF and bFGF are decreased in the tissues of rat pressure ulcer, suggesting that they may be the potential key factors in the difficult healing of pressure ulcer.     

Effects of Shenmai injection on myocardial fibrosis in rat model of diabe-tic cardiomyopathy

AIM：To explore the effects of Shenmai injection on myocardial fibrosis in a rat model of diabetic cardiomyopathy (DCM). METHODS：Wistar rats (n=30) were randomly divided into control group, diabetes group and treatment group. Single intraperitoneal injection of streptozotocin was utilized to establish a rat model of DCM. The rats with DCM in treatment group were intraperitoneally injected with Shenmai injection. Ventricular cannulation was applied to assess the cardiac functions. The formation of collagen in the cardiac tissues was assessed by Masson staining. The generation of reactive oxygen species (ROS) in the cardiac tissues was detected by dihydroethidium staining. The expression levels of matrix metalloproteinase 2 (MMP-2), tissue inhibitor of metalloproteinase 2 (TIMP-2) and collagen I in the cardiac tissues were determined by Western blotting. RESULTS：Compared with control group, the cardiac functions were deteriorated in diabetes group (P<0.05), which was improved in treatment group as compared with diabetes group (P<0.05). Compared with control group, the formation of collagen and ROS increased significantly in diabetes group (P<0.05), which was decreased in treatment group as compared with diabetes group (P<0.05). Compared with control group, the expression level of MMP-2 in the cardiac tissues was deceased and TIMP-2 was increased significantly in diabetes group (P<0.05), but reversed significantly in treatment group (P<0.05).CONCLUSION:Shenmai injection attenuates cardiac fibrosis in the rats with DCM by inhibiting the generation of ROS.     

Effect of puerarin on blood pressure and serum lipid in a rat model of insulin resistance

AIM: To observe the effects of puerarin on blood pressure (BP), serum lipid in a rat model of insulin resistance and the mechanism. METHODS: Adult SD rats were maintained on high-fat-sugar-salt diet for 12 weeks. Puerarin was administered to the rats from 9th week for 4 weeks by intramuscular injection. BP was measured at the end of 0, 8th, 12th week. The levels of serum glucose, serum lipid, fasting serum insulin and the levels of MDA, TNF-α, plasma rennin, angiotensinⅡ (AngⅡ) and the activity of SOD were measured and the insulin-sensitivity index was also calculated at the end of the experiment. RESULTS: High and middle dosage of puerarin significantly decreased blood pressure, reduced the levels of serum lipid and AngⅡ, and also increased insulin-sensitivity index. The levels of MDA and TNF-α were significantly decreased by high dosage of puerarin. The activity of SOD was increased significantly. CONCLUSIONS: Puerarin possesses the effects of decreasing the blood pressure and serum lipid in a rat model of insulin resistance, which may be concerned with the changes of rennin- angiotensin system, the levels of oxygen-derived free radicals and TNF-α.     

Iodoacetamide attenuates cognition impairment and amyloid deposit in an AD rat model

AIM:To investigate the effect of an insulin-degrading enzyme (IDE) activator, iodoacetamide, on an AD rat model. METHODS:Thirty-two male Wistar rats were divided into 4 groups:Aβ group, Aβ+ iodoacetamide group, iodoacetamide group and control. Each group included 8 subjects. Aβ was injected into the CA3 area of bilateral hippocampi;Iodoacetamide was administered subcutaneously daily for 5 consecutive days before surgery. The behavior performance in the morris water maze was obtained. The level of Aβ was detected by immunohistochemistry staining and ELISA assay. RESULTS:Level of Aβ was significantly reduced in Aβ+iodoacetamide group compared with that of Aβ group (dropped by 24%), and the Aβ deposit decreased (by 23.9%), the cognition function of this group also greatly improved according to the behavior performance in the morris water maze. The mean escape latency in the place navigation was much lower compared to Aβ group and the times that rats acrossed the central platform in the spatial probe test was also elevated. CONCLUSION:Iodoacetamide significantly reduces the Aβ deposit in the hippocampus in an AD rat model and relieves memory loss.