1.Function of IL-17 in viral myocarditis mice
Xiaoling ZHANG ; Zonglin YU ; Jinsong REN ; Fuchuan LIU
Chinese Journal of Immunology 2016;32(7):979-982
Objective:To study the effect of IL-17 on viral myocarditis in mice. Methods:We randomly selected 20 wild type BALB/c mice as the control group,20 IL-17A-/- mice for IL-17A-/- group,20 IL-17A-/- mice for IL-17 group. Each mouse had intrap-eritoneal injection of Coxsackie virus B3 (Coxsackie virus B3,CVB3) to construct VMC model. The serum levels of IL-17 were detected by ELISA method,and Th17 cells were detected by flow cytometry. After 3 days of treatment with CVB3,100 μg IgG antibody was injected intraperitoneally in control group and L-17A-/- group,and 100 μg IL-17mAb was injected with IL-17 group. Mice myocardium was collected at 3rd,7th and 14th days,respectively,after CVB3 treatment. Pathological examination was carried out on the myocardial sections of mice and stained with H&E. Virus titer in mouse myocardium was examined. The contents of TNF-α,IL-6,IL-23 and IL-17 in myocardial tissue were detected by enzyme linked immunosorbent assay. Results:We successfully constructed the model of mice with viral myocarditis. The levels of Th17 and IL-17 in serum of IL-17 group were significantly lower than those in control group and IL-17-/-group. On the fourteenth day after CVB3 treatment,the degree of myocardial tissue damage in the control group was significantly higher than that in the IL-17A-/- group and the IL-17 group(P<0. 05),the degree of myocardial injury in IL-17 mice was higher than that in the IL-17A-/- group. The virus titer of the control group was higher than that of IL-17A-/-group and IL-17 group,and the control group was increased with the increase of CVB3 treatment phase (P< 0. 05). After the IL-17mAb was injected,the virus titer of IL-17 group was higher than that of IL-17A-/- group. The levels of IL-17, IL-23, IL-6 and TNF-α in IL-17-/- group and IL-17 group were significantly lower than those in control group. The levels of IL-17,IL-23,IL-6 and TNF-αin IL-17 group were significantly higher than those in IL-17-/-group (P<0. 05). Conclusion:IL-17 is an important inflammatory factor in viral myocarditis,and IL-17 deletion can protect myocardium from viral myocarditis in mice.
2.Therapeutic effect of electroacupuncture plus neural stem cell transplantation on the hindlimb function of rats with spinal cord injury
Jianmin LIU ; Fuchuan WANG ; Yajing ZHOU ; Li MU ; Shaoke HOU ; Lina HAO ; Zitan ZHANG
Chinese Journal of Tissue Engineering Research 2015;(50):8132-8138
BACKGROUND:Neural stem cel transplantation alone has achieved unsatisfactory outcomes in the repair of damaged spinal cord tissues. To promote the survival, proliferation and neuronal differentiation of transplanted cels in vivo, it is necessary to further improve the micro-environment of spinal cord injury.
OBJECTIVE:To investigate the effect of neural stem cel transplantation plus electroacupuncture on the hindlimb function and electrophysiological changes of rats with spinal cord injury.
METHODS: Animal models of spinal cord injury were made in 72 Sprague-Dawley rats and randomized into four groups: control group with injection of culture mediumvia the tail vein; neural stem cel group with injection of neural stem cel suspensionvia the tail vein; electroacupuncture group given 1-week electroacupuncture atDu meridian and body points starting from 6 hours after modeling; combined group given injection of neural stem cel suspension via the tail vein+1-week electroacupuncture atDu meridian and body points starting from 6 hours after modeling. Motor functional recovery in rats was assessed by Basso-Beattie-Bresnahan score and inclined plane test before and at 1, 3 days and 1-4 weeks after modeling. At 4 weeks after modeling, hematoxylin-eosin staining was performed for pathological observation; fluorescence microscope was used to observe the survival and distribution of CM-Dil-labeled neural stem cels; horseradish peroxidase tracer was used to observe nerve fiber regeneration; rat neurophysiological recovery was assessed by determining motor evoked potentials and somatosensory evoked potentials.
RESULTS AND CONCLUSION:At 2-4 weeks after modeling, the hindlimb function was better in the combined group than the neural stem cel group and electroacupuncture group; while it was better in the neural stem cel group and electroacupuncture group than the control group. At 4 weeks after modeling, there were few nerve axon-like structures and smal voids in the spinal cord of the neural stem cel group and electroacupuncture group; however, in the combined group, there were more nerve axon-like structures and no void in the spinal cord. At 4 weeks after modeling, the number of nerve fibers positive for CD-Dil and horseradish peroxidase was ranked as folows: combined group > neural stem cel group and electroacupuncture group > control group, and there were significant differences between groups (P < 0.05). The latencies of motor and somatosensory evoked potentials were significantly lower in the combined group than the neural stem cel group and electroaucpuncture group folowed by the control group (P < 0.05), whereas the amplitudes of motor and somatosensory evoked potentials were significantly higher in the combined group than the neural stem cel group and electroacupuncture group folowed by the control group (P < 0.05). In conclusion, these findings indicate that neural stem cel transplantation combined with electroacupuncture can promote synaptic regeneration and improve the motor and electrophysiological functions of rats.
3.Effect of electroacupuncture stimulation on apoptosis of nerve cells in a rat model of spinal cord contusion
Jianmin LIU ; Fuchuan WANG ; Yajing ZHOU ; Li MU ; Shaoke HOU ; Lina HAO ; Zitan ZHANG
Chinese Journal of Tissue Engineering Research 2016;20(5):616-621
BACKGROUND:Electroacupuncture has promoting effects on the functional recovery of the injured spinal cord, can decrease pain, and elevate postoperative effect after acute spinal cord contusion.OBJECTIVE:To observe the effect of electroacupuncture on apoptosis in the injured site after spinal cord contusion, and analyze its neuroprotective effects on neurological function in rats with spinal cord contusion. METHODS:A total of 66 adult female Sprague-Dawley rats were randomly divided into: sham surgery group (n=20), spinal cord contusion group (n=20), electroacupuncture stimulation group (n=20) because six rats were excluded due to modeling failure and death. Before model establishment, at 1, 3 days, 1, 2, 3 and 4 weeks after model establishment, motor functions were evaluated by BBB score and the inclined plate test. At 3 days after model establishment, apoptosis of nerve cels could be detected in the site of injury in each experimental group using TUNEL assay. mRNA and protein expression of bax, bcl-2 and caspase-3 was detected surrounding the injury site using RT-PCR and western blot assay. Morphological changes in the site of injury could be observed using hematoxylin-eosin staining. The regeneration of nerve fibers was observed using HRP tracing. RESULTS AND CONCLUSION:(1) Motor function score was significantly increased at various time points in the 2nd week of treatment in the electroacupuncture stimulation group than in the spinal cord contusion group (P< 0.05). (2) Apoptotic index was significantly lower in the electroacupuncture stimulation group than in the spinal cord contusion group at 3 days after model establishment (P < 0.05). (3) mRNA and protein expression of bax and caspase-3 was significantly lower in the electroacupuncture stimulation group than in the spinal cord contusion group at 72 hours (P < 0.05); bcl-2 gene and protein expression was significantly higher (P < 0.05). (4) The number of HRP-positive nerve fibers was highest in the sham surgery group, folowed by electroacupuncture stimulation group, and lowest in the spinal cord contusion group at 4 weeks (P < 0.05). Results indicated that electroacupuncture plays a protective role on the spinal cord contusion by reducing apoptosis of nerve cels at the site of injury.