1.Role of activated microglia transplantation in treatment of spinal cord injury in rats:a preliminary study
Tengbo YU ; Dewei KOU ; Yongshuai CHENG ; Yanchen CHU ; Aimin WANG
Chinese Journal of Trauma 2008;24(5):373-377
Objective To study the role of activated microglia transplantation in recovery of hindlimb locomotor function in rats with spinal cord injury(SCI). Methods A total of 40 female adult Wistar rats were selected and divided into four groups randomly(10 rats in each group).The former two groups were locomotor function observation groups,in which rat model with spinal cord was established by striking with improved self-made Allen's strike equipment to fabricate moderate spinal cord injury and divided into transplantation group and control group.The latter two were histological observation groups,the spinal cord injury model was fabricated by the same above-mentioned method and divided into transplantation and control groups.Before fabricating the spinal cord injury model,the microglia of the newborn rats were cultured,separated,purified and identified and the purity of the microglia determined.The injury position was exposed again seven days after transplantation and the cell suspension of microglia was injected around the injury position with microsyringe,which was free in the control group.The hindlimb locomotor function of rats was detected and scored at 1 day,1,2,3 and 4 weeks in the locomotor function observation groups after transplantation respectively.At the same time,two rats were extracted randomly from the control group and the transplantation group in histological observation groups to cut specimen and slice for Naoumenko-Feign paraffin section and dying.Then,the microglia were observed and counted by microscope and analyzed statistically. Results At 1,2,3 and 4 weeks after operation,the BBB score of the control group and the transplantation group was increased gradually with the time.But compared with the control group,transplantation group had higher scores of hindlimb locomotor function at 2,3 and 4 weeks after operation,with statistical difference(P<0.05).Naoumenko-Feigin paraffin section and dying and microglia counting showed that positive microglia number in the transplantation group was increased more obviously than the control group,with statistical difference(P<0.05).Conclusion Activated microglia transplantation can promote the recovery of the hindlimb locomotor function in SCI rats.
2.The VEGF production by dedifferentiated chondrocytes under synovial fluid stimulation from coxarthrosis and femoral neck fracture patients
Tengbo YU ; Yongshuai CHENG ; Kang SUN ; Jinzhao LIU ; Zhijie WANG ; Xuexiao MA ; Aimin WANG
Chinese Journal of Orthopaedics 2010;30(12):1206-1210
Objective To investigate the vascular endothelial growth factor (VEGF) expression level by chondrocytes isolated from patients with osteoarthritis (OA) in hip or femoral neck fracture (FNF) and explore the effect of synovial fluid from OA or FNF on secretion of VEGF. Methods The cartilage tissues were collected from 12 patients with OA in hip and 8 patients with FNF. Cartilage was stained with HIM and Safranin O/Fast Green (S/F) method. The damage of cartilage was evaluated using Mankin scores.Cathepsin B which was selected for cell dedifferentiation monitoring marker and VEGF level was detected in the supernatant fluid. The synovial fluid from OA, FNF and DMEM were respectively added to the culture medium to explore their effects on regulating VEGF. Results Cartilage the Mankin scores of OA group were higher than that of FNF group. Chondrocytes gradually lost their original spherical appearance, with Cathepsin B upregulated while VEGF downregulated. The OA synovial fluid can stimulate chongdrocytes to secrete more VEGF than the one from patients with FNF. However, chondrocytes gradually produced less VEGF after passaging. Conclusion Mankin scores had good correlation with chondrocytes' VEGF production in the early stage of primary culture. Chondrocytes showed quick dedifferentiation characteristics in vitro. OA synovial fluid showed abig ger capability in stimulating chondrocytes to express more VEGF, which might indicate that OA synovial fluid participated in the pathological process of OA.
3.Advances in pharmacological effects of tanshi-none ⅡA on pain
Wenjing CHENG ; Xi CHEN ; Danshen ZHANG ; Yongshuai JING
Chinese Journal of Pharmacology and Toxicology 2023;37(7):524-525
Pain is one of the most serious problems plaguing human health today.Drug therapy is one of the main ways to treat pain in clinic.The analgesic drugs commonly used in clinical treatment of pain are often accompanied by many side effects,the analgesic effect is still not ideal.Salvia miltiorrhiza is a traditional medici-nal material with the same origin as food and medicine.It has the functions of promoting blood circulation and removing blood stasis,relieving pain through menstrual circulation,and contains many effective ingredients such as tanshinone and salvianolic acid.Tanshinone is a kind of rosin diterpenoid compound,which mainly consists of o-quinone type and p-quinone type parent nucleus,and tanshinone Ⅱ A is the representative compound.The pharmacological mechanism of tanshinone ⅡA in labor pain mainly includes:① Regulate inflammatory factors.Inflammatory cytokines played an important role in the occurrence and progression of pain.It was found that the analgesic effect of tanshinone ⅡA was related to the anti-inflammatory effect.Tanshinone ⅡA showed anti-injuri-ous activity in various pain models,such as bone cancer pain and sciatic nerve ligation,and related studies found that tanshinone ⅡA could inhibit the expression of inflammatory factors TNF-α,IL-1β and IL-6 in the spinal cord of model rats.In the spinal nerve ligation model,tanshinone ⅡA also promoted the release of anti-inflam-matory cytokine IL-10 in the spinal cord of rats.② Regu-late signal pathways related to regulating spinal cord oxi-dation and apoptosis.Apoptosis and oxidation played an important role in the process of pain.When nerve injury was caused by stimulation,oxidative stress and apopto-sis of nerve cells were involved in the mechanism of hyperalgesia.Tanshinone ⅡA sodium sulfonate could relieve pain by regulating apoptosis-related pathways.In neuralgia model,tanshinone ⅡA could reduce the apop-tosis of spinal cord neurons by inhibiting oxidative stress response in rat spinal cord tissue.In addition,tanshinone ⅡA also decreased the expression of pro-apoptotic protein in spinal dorsal horn of CCI rats.They included caspase-3,Bcl-2,Bax protein,and enhancer binding protein homologous protein,Increased the expres-sion of anti-apoptosis protein Bcl-2.③ Inhibit the activa-tion of spinal cord glial cells.tanshinone ⅡA could exert its labor pain effect by inhibiting the activation of astro-cytes,including inhibiting the expression of chemo-therapy-induced neuralgia,inflammatory pain and inflam-matory cytokines IL-6,IL-1β and TNF-α,and inhibiting the activation of inflammatory signaling pathways related to astrocyte activation.Such as NF-κB signaling path-way,c-Jun N-terminal kinase signaling pathway,etc.In addition,tanshinone ⅡA also inhibited the activation of microglia by inhibiting the expression of CX3CR1 receptor on the surface of microglia and inhibiting the phosphoryla-tion of ERK,JNK and p38 signaling pathways.④ Decr-ease the expression of glutamate receptors in spinal cord.NMDA is an ionic glutamate receptor in the central nervous system,and its subunit NR2B is closely related to pain.The overexpression of NR2B in spinal cord could lead to the decrease of pain threshold,which was an important mechanism of pain generation.The mechani-cal threshold and thermal threshold of CCI rats were increased by tanshinone ⅡA,and the expression of spi-nal dorsal horn 2B subunit was significantly decreased after tanshinone ⅡA treatment in CCI rats.Therefore,it was concluded that the analgesic effect of tanshinone ⅡA on CCI model may be related to the decreased expres-sion of NR2B in spinal dorsal horn.In conclusion,tanshi-none ⅡA can effectively play the role of labor pain,and has great potential for development in the field of medi-cine and health products.
4.Effect of phage lysin on infection and biofilm formation in rabbit after knee joint prosthesis implantation
Yongshuai CHENG ; Chao QI ; Xia ZHAO ; Bo PANG ; Ken LI ; Tengbo YU
Chinese Journal of Orthopaedics 2018;38(12):724-730
Objective To investigate the effect of local injection of phage lyase LysGH 15 into rabbits' knee joint on the systemic inflammation,local infection around knee joint prosthesis and biofilm formation on the prosthesis surface after their knee joint prosthesis implantation surgeries.Methods Models of Staphylococcus aureus infection on rabbits' knee joint prosthesis after prosthesis implantation were built and divided into experimental group for intra-articular injection with lyase and control group for injection with saline into their joint cavity.The phage lysin LysGH15 was synthesized and purified.On the 1st,2nd and 3rd day after the inoculation with Staphylococcus aureus bacteria into the rabbits' knee joint cavity of the prosthesis implanted side,0.5 ml diluted solution of LysGH15 was injected into the knee joint cavity with infection around the prosthesis for the experimental group rabbits and 0.5 ml saline was injected into the corresponding joint cavity for control group rabbits as blank contrast.On the 1st,3rd,7th and 14th day,blood samples were collected from their ear vein to make plasma procalcitonin test for evaluation of rabbits' systemic infection.After the last time for collection of venous blood samples,these rabbits were killed instantly and their knee joints of prosthesis implantation side were dissociated.Tissue around the prosthesis was processed with HE staining to observe and evaluate the local infection and tissue necrosis around the prosthesis.The biofilm formation on the prosthesis surface was evaluated with semi quantitative method after the observation of samples under scanning electron microscope (SEM).Results After the injection of LysGH15 in experimental group,their serum procalcitonin level,which worked as the systemic inflammatory marker,decreased rapidly especially on the 3rd day after lysin injection.Compared with the control group,the infection degree of experimental group significantly decreased.In the experimental group,the infection and necrosis degree of the tissue surrounding the prosthesis were significantly lower in the experimental group than those in the control group.The semi quantitative scores were conducted for these samples and graded to make rank sum test.The difference between the two groups was statistically significant (U=2.4948,P=0.0126).There was a statistically significant difference in the rank sum test between the two groups in the quality of biofilm formation (U=2.2539,P=0.0242).Conclusion Phage lysin LysGH15 can alleviate the rabbits'systemic inflammation caused by Staphylococcus aureus bacteria after their knee joint prosthesis implantation,reduce the extent of damage caused by infection and inflammation to the tissue around the prosthesis,and inhibit the formation of bacterial biofilm on the surface of implanted prosthesis.