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

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

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

BACKGROUNDMethylprednisolone (MP), a synthetic glucocorticosteroid, has been broadly studied in experiments on endotoxin-induced shock and septic shock. This study was designed to ascertain whether glycine and MP can protect against organ injury and death caused by hemorrhagic shock, and to elucidate the underlying mechanisms of these protective effects in rats.

METHODTo establish a shock model, Wistar rats were bled to maintain mean arterial pressure at 30-50 mmHg for 1 hour and subsequently resuscitated with the shed blood and normal saline. Just prior to resuscitation, the rats were randomly assigned to four groups: sham group (operation performed without inducing shock), shock group, shock + glycine group (glycine injected at the beginning of resuscitation) and shock + MP group (MP injected at the beginning of resuscitation).

RESULTS(1) Seventy-two hours after resuscitation, the survival rate of rats from the shock group had decreased to 20%, while the survival rates of rats from the shock + glycine and shock + MP groups were 77.8% and 80%, respectively. The difference was significant (P <0.05). (2) Eighteen hours after resuscitation, pathological alterations in the organs of the rats were apparent. In rats from the shock group, edema, interstitial leukocyte infiltration, and cellular degeneration occurred in the liver, lungs, kidneys, and heart. Glycine and MP reduced these pathological changes significantly. (3) Eighteen hours after resuscitation, the levels of creatine phosphokinase, transaminases, and creatine were elevated significantly in rats from the shock group, indicating injury to the heart, liver, and kidneys, while these levels were elevated only slightly in the shock + glycine and shock + MP groups. The differences were significant (P <0.01). (4) There were significant increases in intracellular calcium and production of tumor necrosis factor (TNF-alpha) by isolated Kupffer cells stimulated by endotoxin after hemorrhagic shock. These changes were completely prevented by glycine and MP (P <0.01).

CONCLUSIONGlycine and MP reduce organ injury and mortality caused by hemorrhagic shock by preventing increase of intracellular calcium levels in Kupffer cell, suppressing Kupffer cell activation, decreasing the production of TNF-alpha by Kupffer cells, and blocking systemic inflammatory responses.

Animals ; Calcium ; metabolism ; Glycine ; pharmacology ; therapeutic use ; Methylprednisolone ; pharmacology ; therapeutic use ; Multiple Organ Failure ; etiology ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; drug therapy ; mortality ; pathology ; Tumor Necrosis Factor-alpha ; biosynthesis

Although methylprednisolone (MP) is the standard of care in acute spinal cord injury (SCI), its functional outcome varies in clinical situation. Recent report demonstrated that MP depresses the expression of growth-promoting neurotrophic factors after acute SCI. The present study was designed to investigate whether continuous infusion of brain-derived neurotrophic factor (BDNF) after MP treatment promotes functional recovery in severe SCI. Contusion injury was produced at the T10 vertebral level of the spinal cord in adult rats. The rats received MP intravenously immediately after the injury and BDNF was infused intrathecally using an osmotic mini-pump for six weeks. Immunohistochemical methods were used to detect ED-1, Growth associated protein-43 (GAP-43), neurofilament (NF), and choline acethyl transferase (ChAT) levels. BDNF did not alter the effect of MP on hematogenous inflammatory cellular infiltration. MP treatment with BDNF infusion resulted in greater axonal survival and regeneration compared to MP treatment alone, as indicated by increases in NF and GAP-43 gene expression. Adjunctive BDNF infusion resulted in better locomotor test scores using the Basso-Beattie-Bresnahan (BBB) test. This study demonstrated that continuous infusion of BDNF after initial MP treatment improved functional recovery after severe spinal cord injury without dampening the acute effect of MP.
Animals ; Anti-Inflammatory Agents/pharmacology ; Axons/pathology ; Brain-Derived Neurotrophic Factor/metabolism/*pharmacology ; Choline O-Acetyltransferase/metabolism ; Female ; GAP-43 Protein/metabolism ; Gene Expression Regulation ; Immunohistochemistry ; Methylprednisolone/metabolism/*pharmacology ; Osmosis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Spinal Cord/pathology ; Spinal Cord Injuries/*pathology ; Time Factors

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 combination of glycine and methylprednisolone (MP) on Kupffer cells in liver of rats suffered from hemorrhagic shock.

METHODSFifty Wistar rats were bled to establish the shock model and subsequently resuscitated with shed blood and normal saline. Just prior to resuscitation, the rats were randomly assigned to 5 groups: sham group, shock group, shock + glycine group, shock + MP group and shock + glycine + MP group. The intracellular calcium concentration and the level of tumor necrosis factor alpha (TNF alpha) in the culture medium of Kupffer cells were determined after stimulation with different concentrations (1, 10, 100 and 1000 ng/ml) of lipopolysaccharide (LPS).

RESULTSConcentration of intracellular calcium and production of TNF-alpha by isolated Kupffer cells stimulated by LPS were elevated significantly in the rats with hemorrhagic shock, which were totally prevented by glycine + MP compared with other groups (P < 0.005).

CONCLUSIONSThe combination of glycine and MP prevents the increase of intracellular calcium of Kupffer cell, suppress Kupffer cell activation, decrease the production of TNF-alpha of Kupffer cell and block systemic inflammatory responses more effectively than single administer of glycine or MP.

Animals ; Calcium ; metabolism ; Cells, Cultured ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Female ; Glycine ; pharmacology ; therapeutic use ; Kupffer Cells ; drug effects ; pathology ; physiology ; Liver ; drug effects ; metabolism ; pathology ; Male ; Methylprednisolone ; pharmacology ; therapeutic use ; Random Allocation ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; drug therapy ; pathology ; physiopathology ; Tumor Necrosis Factor-alpha ; metabolism

Methylprednisolone (MP), a glucocorticoid steroid, has an anti-inflammatory action and seems to inhibit the formation of oxygen free radicals produced during lipid peroxidation in a spinal cord injury (SCI). However, the effects of MP on the functional recovery after a SCI is controversial. The present study was conducted to determine the effects of MP on the recovery of neural conduction following a SCI. A SCI was produced using the NYU spinal cord impactor. A behavioral test was conducted to measure neurological disorders, and motor evoked potentials (MEPs) were recorded. According to the behavioral test, using BBB locomotor scaling, MP-treated animals showed improved functional recoveries when compared to salinetreated animals. MEP latencies in the MP-treated group were shortened when compared to those in the control group. Peak amplitudes of MEPs were larger in the MP-treated group than those in the control group. The thresholds of MEPs tended to be lower in the MP-treated group than those in the control group. These results suggest that MP may improve functional recovery after a SCI.
Animals ; Disease Models, Animal ; Electrophysiology ; Evoked Potentials, Motor/*drug effects ; Free Radicals ; Glucocorticoids/metabolism ; Male ; Methylprednisolone/*pharmacology ; Neurons/*drug effects ; Oxygen/metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Glucocorticoid/metabolism ; Research Support, Non-U.S. Gov't ; Sodium Chloride/pharmacology ; Spinal Cord/pathology ; Spinal Cord Injuries/*drug therapy ; Time Factors

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