BACKGROUNDPrevention of osteonecrosis (ON) has seldom been addressed. The purpose of this study was to evaluate the effect of resveratrol on preventing steroid-induced ON in rabbits.

METHODSSeventy-two rabbits were divided into four groups: (1) NEC (ON) group: thirty rabbits were treated with lipopolysaccharide (LPS) once, then with methylprednisolone (MPS) daily for 3 days; (2) PRE (prevention) group: thirty rabbits were given one dose of LPS, then MPS daily for 3 days, and resveratrol on day 0 and daily for 2 weeks; (3) RES (resveratrol) group: six rabbits were given resveratrol for 2 weeks but without LPS/MPS; (4) CON (control) group: six rabbits were given alcohol for 2 weeks but without LPS/MPS. Levels of plasma tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor 1 (PAI-1), thrombomodulin (TM), vascular endothelial growth factor (VEGF), maximum enhancement (ME) by magnetic resonance imaging, and ON incidence were evaluated.

RESULTSThe PRE group had a lower ON incidence than the NEC group, but with no significant differences at 2 weeks and 12 weeks. The RES and CON groups did not develop ON. TM and VEGF were significantly higher in the NEC group compared with the PRE group at weeks 1, 2, and 4 (TM: 1 week, P = 0.029; 2 weeks, P = 0.005; and 4 weeks, P = 0.047; VEGF: 1 week, P = 0.039; 2 weeks, P = 0.021; 4 weeks, P = 0.014), but the difference disappeared at 12 weeks. The levels of t-PA and PAI-1 were not significantly different between the NEC and PRE groups. The TM, t-PA, PAI-1, and VEGF concentrations in the RES and CON groups did not change over time. Compared to the baseline, ME in the NEC group decreased significantly (P = 0.025) at week 1, increased significantly (P = 0.021) at week 2, and was decreased at week 12. The variance was insignificant in the PRE group.

CONCLUSIONSResveratrol may improve blood supply to bone in a rabbit model of ON of the femoral head via anti-inflammatory effects to protect the vascular endothelium and reduce thrombosis.

Animals ; Disease Models, Animal ; Femur Head Necrosis ; chemically induced ; prevention & control ; Lipopolysaccharides ; toxicity ; Magnetic Resonance Imaging ; Methylprednisolone ; toxicity ; Plasminogen Activator Inhibitor 1 ; blood ; Rabbits ; Stilbenes ; pharmacology ; therapeutic use ; Thrombomodulin ; blood ; Tissue Plasminogen Activator ; blood ; Vascular Endothelial Growth Factor A ; blood

OBJECTIVETo study different effects of Herba Lycopodii (HL) Alcohol Extracted Granule combined methylprednisolone on behavioral changes, brain derived neurotrophic factor (BDNF) expression levels, and N-methyl-D-aspartate (NMDA) receptor levels in rats with spinal cord injury (SCI).

METHODSMale adult SD rats were randomly divided into five groups, i.e., the sham-operation group, the model group, the HL treatment group, the methylprednisolone treatment group, the HL + methylprednisolone treatment group. Rats in the HL treatment group were intragastrically administered with HL at the daily dose of 50 mg/kg for 5 successive days. Rats in the methylprednisolone treatment group were intramuscularly injected with 50 mg/kg methylprednisolone within 8 h after spinal cord contusion, and then the dose of methylprednisolone was reduced for 10 mg/kg for 5 successive days. Rats in the HL + methylprednisolone treatment group received the two methods used for the aforesaid two groups. Basso Beattie and Bresnahan (BBB) score (for hindlimb motor functions) were assessed at day 0, 3, 7, and 28 after operation. At day 13 after SCI, injured spinal T8-10 was taken from 8 rats of each group and stored in liquid nitrogen. The N-methyl-D-aspartate (NMDA) receptor affinity (Kd) and the maximal binding capacity (Bmax) were determined using [3H]MK-801 radioactive ligand assay. Rats' injured spinal cords were taken for immunohistochemical assay at day 28 after SCI. Expression levels of BDNF in the ventral and dorsal horn of the spinal cord were observed.

RESULTSCompared with the sham-operation group, the number of BDNF positive neurons in the ventral and dorsal horn of the spinal cord increased in the model group, Bmax increased (470 ± 34), Kd decreased, and BBB scores decreased at day 3 -28 (all P <0. 05). Compared with the SCI model group, the number of BDNF positive neurons and Kd increased, BBB scores at day 3 -28 increased (P <0. 05) in each medicated group. Bmax was (660 ± 15) in the methylprednisolone treatment group, (646 ± 25) in the HL treatment group, and (510 ± 21) in the HL +methylprednisolone treatment group (P <0. 05). Compared with the methylprednisolone treatment group, the number of BDNF positive neurons and Kd increased, BBB scores at day 7 -28 increased, and Bmax decreased in the HL treatment group and the HL + methylprednisolone treatment group (all P <0. 05). Compard with the HL treatment group, the number of BDNF positive neurons and Kd increased, and Bmax decreased (all P < 0.05).

CONCLUSIONSHL could effectively improve motor functions of handlimbs, increase expression levels of BDNF in the spinal cord, and lessen secondary injury by affecting spinal levels of NMDA receptors. It showed certain therapeutic and protective roles in treating SCI. Its effect was better than that of methylprednisolone with synergism.

Animals ; Brain-Derived Neurotrophic Factor ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Ethanol ; Male ; Methylprednisolone ; pharmacology ; therapeutic use ; Models, Animal ; N-Methylaspartate ; metabolism ; Neurons ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; Spinal Cord Injuries ; drug therapy ; metabolism

OBJECTIVETo study the effects of a Gold Belt (GB, a traditional Chinese herbal medicine) combined with methyl-prednisolone (MP) on the motor function and brain-derived neurotrophic factor (BDNF) expression in rats with contusive spinal cord injury (SCI).

METHODSThirty adult female SD rats were randomly divided into 5 equal groups, namely the sham-operated group, SCI group, SCI with MP treatment group (MP group, with intramuscular injection of 50 mg/kg MP within 8 hours after SCI and then dosage reduced 10 mg/kg daily), SCI with GB treatment group (GB group, with intragastric gavage of GB 50 mg/kg once daily for 7 days), and combined GB and MP treatment group. The Basso, Beattie and Bresnahan (BBB) locomotor scale was used to evaluate the hindlimb motor function of the rats on days 1, 3, 7, 14, 21 and 28 after the injury. After the last evaluation the rats were sacrificed for immunohistochemistry to observe the localization of BDNF in the ventral and dorsal horn of spinal cord.

RESULTSBDNF were distributed mainly in neurons in the spinal cord grey matter ventral horn and dorsal horn of the rats. The number of BDNF-positive neurons and BBB scores in the combined treatment group were significantly higher than those in the other 4 groups (P<0.05).

CONCLUSIONGB combined with MP produces better therapeutic effects for treating SCI than GB or MP used alone, and such effects are probably related with enhanced BDNF expression in the spinal cord.

Animals ; Brain-Derived Neurotrophic Factor ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Immunohistochemistry ; Methylprednisolone ; pharmacology ; Neurons ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; drug therapy ; metabolism

OBJECTIVE: To explore the effect of methylprednisolone on bone mass, microarchitecture and microdamage in cortical bone of ulna in rats. METHODS: Twenty female Sprague-Dawley rats (3.5 months old) were randomly assigned to two groups: a treatment group and a control group (n=10 per group). The treatment group was subcutaneously injected with methylprednisolone 3.5 mg/(kg.d) while the control group was subcutaneously injected with same volume of vehicle (saline). Rats were sacrificed at 9 weeks after the treatments. Before the sacrifice, the body weight and total bone mineral density (BMD) were measured. The right forelimb was separated through humeral shoulder and then single axial fatigue loading was performed on the right ulna. After fatigue load, the middle ulna section was bulkstained in basic fuchsin. Bone histomorphometry and microdamage analysis were performed on the middle ulna section. RESULTS: Compared with the control group, the body weight, total bone BMD and ulnas BMD in the treatment group were decreased by 15%, 6.4% and 4.3% respectively (all P<0.05); the ulna inner perimeter and marrow area in the treatment group were increased by 23.3% and 32%, respectively (both P<0.05), while the outer perimeter were decreased by 3.1% (P>0.05). There was no significant difference in the cortical and total area between the 2 groups (both P>0.05). The number of microcrack, microcrack density and microcrack surface density in the treatment group were increased by 43%, 48% and 50%, respectively, compared with those in the control group (all P<0.05), but there was no significant difference in the mean length of microcrack between the 2 groups (P>0.05). CONCLUSION Methylprednisolone can significantly induce the bone loss and the deterioration of microarchitecture and microdamage in ulna of rats.
Animals ; Bone Density ; drug effects ; Female ; Methylprednisolone ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Ulna ; drug effects ; pathology

OBJECTIVE: To determine the influence of stress on myocardial apoptosis in ischemic preconditioning group (IPC). METHODS: Twenty-four Japanese white rabbits were randomly divided into 4 groups (n=6): an etomidate group (the Etom group) of depressed stress established by intravenous etomidate, an IPC group, an ischemic reperfusion group (the IR group) and a methylprednisolone group (the MP group). Myocardial apoptosis was examined by DNA-laddering, in situ nick-end labeling (TUNEL) and Hoechst dyeing. RESULTS: The DNA ladder increased in the Etom group. The percentage of apoptosis by TUNEL method was 1.7%±0.2% in the IPC group, 2.3%±0.8% in the MP group, 3.8%±1.3% in the IR group and 3.0%±0.4% in the Etom group. Hoechst dying was 4.1%±0.9% in the IPC group, 3.5%±0.4% in the MP group, 6.2%±1.6% in the IR group and 7.6%±0.4% in the Etom group. There was significant difference between the IPC group and the Etom group or IR group, and also between the MP group and the IR group. CONCLUSION A depressed stress response impairs the inhibition on myocardial apoptosis in ischemic preconditioning. Methylprednisolone may inhibit myocardial apoptosis.
Animals ; Apoptosis ; Etomidate ; pharmacology ; Heart ; drug effects ; Ischemic Preconditioning ; Ischemic Preconditioning, Myocardial ; Methylprednisolone ; pharmacology ; Myocardium ; pathology ; Rabbits

OBJECTIVETo investigate the effect of prostaglandins E1 combined with Xuebijing injection on the expression of transforming growth factor-β₁ (TGF-β₁) and tumor necrosis factor-α (TNF-α) in rats with acute pulmonary interstitial fibrosis.

METHODSA rat model of pulmonary interstitial fibrosis was established by intratracheal injection of bleomycin (1 ml/kg). One hundred and eight Wistar rats were randomly divided into six groups with 18 in each group, which were normal control group, model group, hormone (methylprednisolone) treatment group, Xuebijing treatment group, prostaglandin E1 treatment group and combination treatment group (prostaglandin E1 and Xuebijing injection). Except for those in the normal control group, the rats in each group were sacrificed on the 7th, 14th and 28th day after treatment. The TGF-β₁ expression in lung tissue was measured by immunohistochemical staining. The TNF-α concentration in bronchoalveolar lavage fluid (BALF) of rat model was determined by enzyme-linked immunosorbent assay.

RESULTSThe combination treatment group showed significantly more macrophages with TGF-β₁ expression in lung tissue at each time point, as compared with the model group, Xuebijing treatment group, methylprednisolone treatment group and prostaglandin E1 treatment group (P < 0.05). On the 7th day, the TNF-α concentration in BALF in the combination treatment group was significantly lower than those in the model group, methylprednisolone treatment group and prostaglandin E1 treatment group (P < 0.05); on the 14th day, the TNF-α concentration in BALF in the combination treatment group was significantly lower than that in the model group (P < 0.05); on the 28th day, the levels of TNF-α in the prostaglandin E1 treatment group and combination treatment group were significantly lower than that in the model group (P < 0.05).

CONCLUSIONProstaglandin E1 combined with Xuebijing injection may significantly inhibit TGF-β₁ expression in the lung tissue of rats with acute pulmonary interstitial fibrosis, which reduces alveolar inflammatory response.

Alprostadil ; administration & dosage ; pharmacology ; Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Female ; Male ; Methylprednisolone ; administration & dosage ; pharmacology ; Pulmonary Fibrosis ; metabolism ; pathology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Transforming Growth Factor beta1 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo observe the changes of human umbilical venous endothelial cells (HUVECs) induced by the sera from children with active Henoch-Sch-nlein purpura (HSP) and the protective effects of methylprednisolone against HUVECs injury.

METHODSHUVECs were divided into four groups based on the culture conditions: blank control group, normal serum group, HSP serum group, and HSP serum plus methylprednisolone group. The levels of tumor necrosis factor (TNF)-α and interleukin (IL)-8 in the supernatants of each group were detected using ELISA and the nitric oxide (NO) level by nitrate reductase determination. Moreover, the expressions of nuclear factor-kappa B (NF-κB) and Fractalkine in HUVECs were examined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot, respectively.

RESULTSThe levels of IL-8, TNF-α, and NO in the HSP serum group were significantly higher than those in the blank control and normal serum groups (P<0.05). Compared with the HSP serum group, the levels of IL-8, TNF-α, and NO in the HSP serum plus methylprednisolone group decreased significantly (P<0.05). The mRNA expression levels of NF-κB and Fractalkine in the HSP serum group were significantly higher than those in the blank control group (P<0.05). The protein expression levels of NF-κB and Fractalkine in the HSP serum group were significantly higher than those in the blank control and normal control group (P<0.05). Compared with the HSP serum group, the mRNA and protein expression levels of NF-κB and Fractalkine in the HSP serum plus methylprednisolone group decreased significantly (P<0.05).

CONCLUSIONSThe sera from children with active HSP can induce the in vitro cultured HUVECs to become activated and excrete cytokines. Methylprednisolone may inhibit NF-κB expression, reduce the production of inflammatory factors, and thus alleviate vascular inflamation.

Cells, Cultured ; Chemokine CX3CL1 ; analysis ; genetics ; Child ; Child, Preschool ; Cytokines ; blood ; Cytoprotection ; Female ; Human Umbilical Vein Endothelial Cells ; drug effects ; immunology ; metabolism ; Humans ; Male ; Methylprednisolone ; pharmacology ; NF-kappa B ; analysis ; antagonists & inhibitors ; genetics ; Nitric Oxide ; physiology ; Purpura, Schoenlein-Henoch ; blood

OBJECTIVETo investigate the effect of two treating principles and formulas, which are named 'invigorating spleen to remove phlem and promoting blood circulation to remove meridian obstruction' (Jianpi) and 'invigorate the kidney and promoting blood circulation to remove meridian obstruction' (Bushen), on the expression of osteogenic factors in steroid-induced osteonecrosis of the femoral head (SONFH), such as bone morphogenetic protein 2 (BMP2) and transforming growth factor beta1 (TGFbeta1) and Smads, as well as to explore and compare their mechanisms of prevention and treatment of SONFH.

METHODAnimal model of SONFH was established by injection with methylprednisolone in chest muscle on chickens. 48 SONFH chickens were randomly assigned to model, Jianpi and Bushen group. Another 16 normal chickens served as control group. At the 8th and 16th week, the expression of BMP2, TGFbeta1, Smad4 and Smad7 of bilateral femoral heads were detected with immunohistochemistry.

RESULTThe expression of BMP2, TGFbeta1 and Smad4 decreased, and Smad7 increased significantly in model group compared with control group. The expression of BMP2, TGFbeta1, Smad4 increased and Smad7 decreased significantly in Jianpi group at the 8th week compared with model group, and the same changes in Bushen group at the 16th week.

CONCLUSIONBoth Jianpi and Bushen formulas exerted preventive and therapeutic activity on SONFH through regulating the expression of BMP2, TGFbeta1, Smad4 and Smad7 to promote bone repair. Notably Jianpi formula took effect earlier than Bushen formula

Animals ; Bone Morphogenetic Protein 2 ; metabolism ; Chickens ; Drugs, Chinese Herbal ; pharmacology ; Female ; Femur Head Necrosis ; chemically induced ; metabolism ; pathology ; Methylprednisolone ; Osteogenesis ; drug effects ; Random Allocation ; Smad4 Protein ; metabolism ; Smad7 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo investigate the effects of methylprednisolone pretreatment on pulmonary lung permeability index and the content of the pulmonary surfactant dipalmitoylphosphatidylcholine (DPPC) in a rabbit model of reexpansion pulmonary edema.

METHODSTwenty-one male New Zealand white rabbits were randomly divided into control group, reexpansion, and reexpansion+methylprednisolone pretreatment groups. The rabbit model of reexpansion pulmonary edema was established using Sakaos method. A bolus dosage of methylprednisolone (3 mg/kg) in reexpansion+methylprednisolone group group or 2.0 ml/kg normal saline in the other two groups was administered intravenously 20 min before reexpansion pulmonary edema. Bronchoalveolar lavage fluid (BALF) and arterial blood samples were collected for measurement of the total protein (TP) and DPPC contents 4 h after reexpansion, and the pulmonary permeability index was calculated.

RESULTSThe pulmonary permeability index in methylprednisolone pretreatment group was significantly lower than that in the reexpansion group (0.007∓0.002 vs 0.177∓0.004, P<0.05). Methylprednisolone pretreatment significantly increased DPPC concentration in the BALF as compared with saline treatment in the reexpansion group (61.815∓28.307 vs 101.955∓24.544 µg/ml, P<0.05).

CONCLUSIONMethylprednisolone pretreatment can increase pulmonary surfactant content and improve pulmonary permeability in the rabbit model of reexpansion pulmonary edema.

1,2-Dipalmitoylphosphatidylcholine ; analysis ; Animals ; Bronchoalveolar Lavage Fluid ; Capillary Permeability ; drug effects ; Male ; Methylprednisolone ; pharmacology ; Permeability ; Pulmonary Edema ; metabolism ; physiopathology ; Pulmonary Surfactants ; metabolism ; Rabbits

Dexamethasone ; therapeutic use ; Glucocorticoids ; pharmacology ; therapeutic use ; Humans ; Infant, Newborn ; Meconium Aspiration Syndrome ; drug therapy ; physiopathology ; Methylprednisolone ; therapeutic use ; Prednisolone ; therapeutic use