ABSTRACT
ObjectiveTo explore the effects and possible mechanism of Wenshen Tongdu Formula （温肾通督方， WTF） on spinal cord injury. MethodsThirty-six C57BL/6 female mice were randomly divided into sham operation group， model group and WTF group， with 12 mice in each group. The spinal cord injury model was established in the model group and the WTF group using the modified Allen's method， while in the sham operation group the spinal cord was only exposed. Since the 1st day after surgery， 50 g/（kg·d） of WTF solution was given to the WTF group by gavage， while 20 ml/（kg·d） of normal saline was given to the sham operation and model group by gavage， all for 14 days. Before surgery and on the 1st， 7th， and 14th days after surgery， the motor function of the mice was evaluated using the inclined plane test and hind limb motor function score （by BMS）. On the 3rd day after surgery， the nerve electrophy-siology was detected through electromyography and motor evoked potential； the spleen length was measured， and B cells in the spleen were sorted by magnetic beads； the differential expression of proteins were detected through proteomics technology； and the protein expression of mitochondrial outer membrane transport porin 20 （Tom20） and downstream cleaved caspase-3 in spleen B cells were measured using Western blotting. On the 14th day after surgery， MRI was used to observe the recovery of the spinal cord. ResultsCompared to those in the sham operation group at the same time， the BMS scores and subscores and the inclined plane test angle in the model group were reduced on the 1st， 7th and 14th days after surgery； the peak value of electromyogram and motor evoked potential were reduced， and the spleen length was shortened， while the expression of Tom20 and cleaved caspase-3 increased in splenic B cells increased （P＜0.05）. Compared to those in the model group at the same time， the BMS subscores on the 14th day and the angle of the inclined plane test on the 7th and 14th days after surgery increased in the WTF group； the peak value of electromyography and motor evoked potential， as well as the length of spleen increased， and the expression of Tom20 and cleaved caspase-3 decreased （P＜0.05）. The proteomics results showed that there were 100 differential proteins in the WTF group versus the model group， of which 37 were up-regulated and 63 were down-regulated. GO enrichment analysis showed that differential proteins mainly played their roles in oxygen binding， exogenous apoptosis negative feedback， zinc ion response， and oxygen transport. KEGG enrichment analysis showed that differential proteins were mainly concentrated in metabolic pathways， Huntington's disease， oxidative phosphorylation and other pathways. Subcellular localization showed that differential proteins were associated with mitochondria. Magnetic resonance imaging on the 14th day after surgery showed that the spinal cord structure of the mice in the sham operation group was intact， and the segments were clear， with normal spinal cord signal； the low signal area in the spinal cord injury area increased in the model group， and the spinal cord became significantly thinner； the injured segment had obvious depression in the WTF group， but the structure was more complete than that in the model group. ConclusionWTF may promote spinal cord injury repair by regulating immune function， and its mechanism may be related to inhibiting pyroptosis of spleen B cells.
ABSTRACT
AIM: To observe the effect of Chinese medicine compound JiSuiKang on the microglia infiltration and neuron survival at the spinal cord injury area of rats. METHODS: Sixty SD rats were randomly divided into sham operation group (Sham), model group (Ctrl), JSK low (JSK-L), JSK medium (JSK-M) and JSK high (JSK-H) dose group, prednisone group. The model of rat spinal cord injury (SCI) was established by improved Allen's method. After the operation, each group was intervened in different ways, and then we assessed the double hind limb motor function score in rats (BBB score). The heart perfusion was done and the spinal cord tissues from the injury area were taken. ELISA tests were used to detect the expression of inflammatory factors. The spinal cord tissue was sliced into sections and then immunofluorescence stained of CD11b/c and beta 3-Tubulin to observe the infiltration of microglia and the growth of neurons under the inverted fluorescence microscope. RESULTS:The BBB score of Sham, JSK-L, JSK-M, JSK-H and prednisone group were higher than Ctrl(P0.05). For the growth state of the neuron, the JSK-L, JSK-M, JSK-H, prednisone group were better than Ctrl (P<0.01), and JSK-H was better than JSK-L and JSK-M (P<0.05). CONCLUSION: The Chinese medicine compound JSK inhibits the migration and invasion of microglia in SCI, promotes the recovery of damaged neurons and motor function in SCI rats.
ABSTRACT
As a kind of mechanical effector cells, chondrocytes can produce a variety of physical and chemical signals under the stimulation of multiaxial load
Subject(s)
Apoptosis , Cells, Cultured , Chondrocytes , Stress, MechanicalABSTRACT
After the articular cartilage injury, the metabolic level is increased during the progressive degeneration, the chondrocytes secrete a variety of inflammatory factors, and the original cell phenotype is gradually changed. For a long time, a large number of researchers have done a lot of researches to promote anabolism of chondrocytes and to maintain the stability of chondrocyte phenotype. There are many molecular signaling pathways involved in the process of promoting cartilage repair. This review focuses on the key signaling molecules in articular cartilage repair, such as transforming growth factor-beta and bone morphogenetic protein, and reveals their roles in the process of cartilage injury and repair, so that researchers in related fields can understand the molecular mechanism of cartilage injury and repair widely and deeply. Based on this, they may find promising targets and biological methods for the treatment of cartilage injury.
Subject(s)
Humans , Bone Morphogenetic Proteins , Physiology , Cartilage, Articular , Wounds and Injuries , Chondrocytes , Physiology , Regeneration , Signal Transduction , Transforming Growth Factor beta , PhysiologyABSTRACT
Neurogenic bowel dysfunction (NBD) manifested as constipation and fecal incontinence often occurs after spinal cord injury (SCI).NBD affects patients' quality of life and is an urgent clinical problem to be solved.The mechanism of NBD is related to central and autonomic nervous system dysfunction,intestinal nervous system dysfunction,changes in intestinal microorganism composition and abnormal content of neurotransmitters.The evaluation method of NBD is mainly based on scoring and imaging,which lacks unified criteria,and the treatment method for NBD is the combination of traditional Chinese and Western medicine.The author summarizes the mechanism,evaluation method,treatment and nursing of NBD in order to provide new insight into these aspects to improve clinical efficacy.
ABSTRACT
This study was aimed to investigate the effect of traditional Chinese medicine (TCM) compound "Ji-Sui-Kang (JSK)" on the differentiation of neural stem cells (NSCs).Drug serum of JSK was prepared.The primary NSCs were isolated and cultured.NSCs were identified.Fetal bovine serum (FBS) was used to induce differentiation of NSCs.Different doses of drug serum of JSK was added to intervene the differentiation.There were the high-,medium-,and low-dose group.The blank control group was also set.The β Ⅲ-tubulin and GFAP were used to detect the differentiation of NSCs.And then,it was compared with the control group without drug serum.Positive cells ofβ m-tubulin and GFAP in different visual fields were counted.And the percentage of positive cells in the total number of cells was calculated.Fluorescence microscope was used to conduct quantitative fluorescence experiments.The results showed that the cultured neurospheres were expressed of Nestin protein.The percentage ofβⅢ-tubulin positive cells in the high-,medium-and low-dose group of drug serum of JSK was significantly higher than that in the control group (P＜0.05).The percentage of GFAP positive cells in the high-,medium-and low-dose groups of drug serum of JSK was significantly lower than that in the control group (P＜0.05).And the high-dose group of drug serum of JSK was statistically significant compared with the middle-and low-dose group of drug serum of JSK (P＜0.01).It was concluded that TCM compound JSK promoted thedifferentiation of NSCs intoβⅢ-tubulin positive cells.It provided a basis for treatment of SCI with TCM.
ABSTRACT
BACKGROUND: Myelin sheath related inhibitors have been found to have great impact on microenvironment of axon regeneration. Traditional Chinese medicine is gradual y becoming a research hotspot on improving microenvironment of nerve regeneration with its advantage on multiple factors and targets. OBJECTIVE: To clarify the effect of Jisuikang on Nogo-66 receptor NgR expression after spinal cord injury. METHODS: 144 rats were randomly divided into six groups: sham surgery group, model group, prednisone group, high-, moderate- and low-dose Jisuikang groups (n=24). Animal models of spinal cord injury were established by the modified Allen’s method in the later five groups. Rats in the prednisone group were daily given 0.06 g/kg prednisone acetate by lavage, once a day. Rats in the high-, moderate- and low-dose Jisuikang groups were daily intragastrically given 12.5, 25 and 50 g/kg Jisuikang, once a day. Rats in the sham surgery and model groups were intragastrically daily given 20 mL of saline, once a day. Rats in each group were administered drugs until death. RESULTS AND CONCLUSION: Compared with the model group, NgR protein and mRNA expression levels were significantly lower in the prednisone, moderate-and low-dose Jisuikang groups. These data suggested that Jisuikang can improve the recovery of neurological function after spinal cord injury and effectively inhibit NgR protein expression at the site of injury so as to suppress the microenvironment factors harmful to nerve regeneration and further improve the microenvironment of nerve regeneration. Subject headings: Drugs, Chinese Herbal; Spinal Cord Injuries; Axons; Tissue Engineering
ABSTRACT
Objective To clarify the effects of Jisuikang on expression levels of BDNF and NGF protein in serum of rats with spinal cord injury;To explore its probable mechanism to improve the axon regeneration microenvironment and resistance spinal cord injury.Methods Ninety SD rats were chosen to establish acute spinal cord injury model through modified Allen's method. 15 rats were chosen randomly as sham-operation group, and the other 75 rats were randomly divided into model group, prednisone group and high-, medium-, and low-doseJisuikang groups. All administration groups were given relevant medicine for gavage, while rats in sham-operation group and model group were given normal saline with the same volume for gavage. The activity and death condition of rats were recorded. Rats were put to death on the 3rd, 7th and 14th d of intervention for carotid artery blood collection. The expressions of BDNF and NGF in serum were detected by ELISA.Results After modeling, the rats in model group showed a typical paraplegia syndrome. 3 day after operation, BDNF showed no statistical significance between model group and prednisone group, while each treatment group showed significant difference when compared with prednisone group (P<0.05,P<0.01). 7 d and 14 d after operation, compared with the model group, BDNF of prednisone group and medium-dose Jisuikang group showed statistical significance, and there was no significant difference between prednisone group and medium-dose Jisuikang group (P<0.05,P<0.01). The expression of NGF in each treatment group at each time point with the prednisone group showed statistical significance. 3 d and 7 d after operation, among prednisone group, high- and medium-doseJisuikang groups, the expression of NGF was significantly higher than model group. 14 d after operation, compared with the model group, the expression of NGF in medium- and low-dose Jisuikang groups and prednisone group still maintained at a relatively high level, with statistical significance (P<0.05,P<0.01).Conclusion Jisuikang can effectively promote the expressions of BDNF and NGF in serum after spinal cord injury and maintained at a relatively high level, so as to improve the microenvironment of axonal regeneration and promote nerve regeneration.
ABSTRACT
Objective To explore the effect of Jisuikang on neural functional recovery, and expression of brain-derived neurotrophic fac-tor (BDNF) protein and mRNA level after spinal cord injury (SCI). Methods 144 female Sprague-Dawley rats, weighted 180 to 220 g, were used for experiment. 24 rats were randomly extracted into sham group (Group A), which had their vertebral plates and spines bitten away on-ly. The others were randomly divided into model group (Group B), prednison group (Group C), and high, middle and low doses of Jisuikang group (Groups D to F) after SCI, 24 rats in each group. Group C was given 0.06 g/(kg?d) prednison, and Groups D to F were given 50, 25 and 12.5 g/(kg?d) Jisuikang respectively, which were given 20 ml/(kg?d) volume by intragastric administration. Groups A and B were given the same volume of normal saline (NS). The Basso-Beattie-Bresnahan (BBB) scores and oblique board test were applied to test the postoper-ative results 24 hours, 3, 7 and 14 days after SCI. The rats were executed and the spinal cord tissues were extracted 3, 7 and 14 days after SCI. Immunohistochemistry, Western blotting and RQ-PCR were applied to test the expression of protein and mRNA of BDNF. Results BBB scores and angle of oblique board test were significantly lower in Groups B to F than in Group A 24 hours after SCI (P0.05). The results of RQ-PCR showed that prednisone and Jisuikang promot-ed the expression of BDNF mRNA. Group C (prednisone) had a most obvious effect at the beginning while Group E was better than Group C 14 days after SCI. Conclusion Jisuikang can promote the neural functional recovery and the expression of BDNF on both protein and mRNA level in SCI rats.