1.Effect of 3-N-butylphthalide pretreatment on the score of neurological deficit, oxidative stress and pathomorphology in rats with cerebral ischemia reperfusion injury
Hairu JI ; Lingwei KONG ; Wei KONG ; Shumin ZHAO ; Xiaoying ZHENG ; Meng CHEN
Acta Anatomica Sinica 2014;(5):622-626
Objective To investigate the effects of 3-N-butylphthalide ( NBP ) pretreatment on the score of neurological deficit , oxidative stress and pathomorphology in rats with cerebral ischemia reperfusion injury ( CIRI ) . Methods Ninety male SD rats were randomly divided into sham operation group ( Sham group ) , model group ( IR group), NBP pretreatment low dose group (NBPⅠgroup), NBP pretreatment middle dose group(NBPⅡgroup) and NBP pretreatment high dose group(NBPⅢgroup), 18 rats per group.Pretreatment was given once a day within 1 week before establishing the model of cerebral ischemia reperfusion injury .The model of middle cerebral artery occlusion ( MCAO) was subjected by suture method .The score of neurological deficit was executed after ischemia for 2h and reperfusion for 24h in all the rats.The cerebral infarction was observed by TTC staining .The pathologic change of brain was observed by HE staining under the microscope .Hydroxylamine method was used to detect activity of SOD , chemical colorimetry method was used to measure activity of GSH-PX, and TBA method was used to detect content of MDA .Results (1) In Sham group, the score of neurological deficit and the percentage of infarction volume were zero , the morphology of nerve cell was regular , and activity of SOD, GSH-PX and content of MDA of brain tissue were normal .(2) Compared with IR group , the score of neurological deficit was significantly reduced in NBP pretreatment groups (all P<0.01); the score of neurological deficit was decreased progressively in turn in NBP Ⅰ,Ⅱ,Ⅲgroup (all P<0.05).(3) Compared with IR group, the percentage of infarction volume was cut down progressively in turn in NBPⅠ,Ⅱ,Ⅲgroup (all P<0.05), and neuron injury was also induced obviously in NBP pretreatment groups .(4) Activity of SOD, GSH-PX was largely increased , and content of MDA was greatly decreased in NBP pretreatment groups ( P<0.01 ) .Activity of SOD , GSH-PX went up progressively in turn , and contents of MDA were cut down progressively in turn in NBP Ⅰ,Ⅱ,Ⅲgroup ( all P<0.05 ) .Conclusion 3-N-butylphthalide can significantly up-regulate the activity of SOD and GSH-PX, decrease the content of MDA , reduce the percentage of infarction volume , and relieve the damage of nerve cell to preventively protect the rats with cerebral ischemia reperfusion injury .
2.Effects of 3-n-butylphthalide pretreatment on microvascular architecture and free radical metabolism in hippocampus induced by cerebral ischemia/reperfusion
Xiaoying ZHENG ; Wei KONG ; Shumin ZHAO ; Hairu JI ; Meng CHEN ; Weijun MA ; Li HAN ; Sheng LIU
Tianjin Medical Journal 2015;(8):867-870
Objective To observe microvascular architecture and free radical metabolism in hippocampus after focal cerebral ischemia/reperfusion and to explore the effect of NBP (3-n-butylphthalide). Methods Fifty-four SD rats were ran?domly divided into NBP pretreatment group, ischemia/reperfusion group and sham operation group (n=18 in each group). The model of middle cerebral artery occlusion(MCAO)was established by suture method. The neurological scores were counted and the volume of infarction was measured;TA-Fe method was applied to observe the microvascular architecture of hippo?campus, Mivnt image analysis system was used to analyze the microvessel density(MVD)and the microvessel area density (MVA)of hippocampus quantitatively;The activity of SOD and content of MDA were measured by colorimetric method. Re?sults Compared to the ischemia reperfusion(IR)group, the neurological scores and the volume of infarction were decreased sharply in NBP group. What′s more, the activity of SOD, MVD and MVA were all enhanced but the content of MDA and the count of closed microvessels were both reduced(P < 0.01). Conclusion NBP can improve microvascular architecture of hippocampus and reduce the free radical injury. There is a protective effect on hippocampus of rats who suffered focal cere?bral ischemia reperfusion.
3.The correlation between lipid peroxide, total oxidation state, apolipoprotein(a) and pregnancy outcome of polycystic ovary syndrome in vitro fertilization-embryo transfer
Jiao WANG ; Hairu SUN ; Xuexia JI
Chinese Journal of Postgraduates of Medicine 2022;45(12):1071-1075
Objective:To investigate the relationship of lipid peroxide (LPO), total oxidation state (TOS), apolipoprotein (a) [apolipoprotein(a), Apo(a)] and pregnancy outcome in patients with polycystic ovarian syndrome (PCOS) treated by ovulation induction-in vitro fertilization-embryo transfer (IVF-ET).Methods:The clinical of 215 patients with PCOS treated by IVF-ET who were admitted to Hengshui People′s Hospitalfrom May 2017 to February 2020 were collected and they were divided into clinical pregnancy group (155 cases) and biochemical pregnancy group (60 cases) according to pregnancy outcome. The levels of LPO, TOS, and Apo(a) in the peripheral blood of the two groups were detected and the data were analyzed.Results:The levels of LPO and TOS before ovulation induction and human chorionic gonadotropin (HCG) dayin the biochemical pregnancy group were higher than those in the clinical pregnancy group: (10.35 ± 3.67) μmol/L vs. (7.16 ± 1.59) μmol/L, (17.98 ± 3.15) mmol H 2O 2 equiv/L vs. (15.03 ± 3.21) mmol H 2O 2 equiv/L; (12.81 ± 4.09) μmol/L vs. (7.38 ± 2.14) μmol/L, (19.66 ± 3.02) mmol H 2O 2 equiv/L vs. (15.19 ± 3.34) mmol H 2O 2 equiv/L; and the level of Apo(a) was lower than that in the clinical pregnancy group: (379.8 ± 95.9) mg/L vs. (486.5 ± 100.3) mg/L, (335.8 ± 84.7) mg/L vs. (473.5 ± 112.9) mg/L, the differences were statistically significant ( P<0.05). LPO and TOS before ovulation induction and HCG day were negatively correlated with the number of high-quality embryos ( P<0.01), and Apo(a) was positively correlated with the number of high-quality embryos ( P<0.01). The risk of non-clinical pregnancy for those with LPO, TOS, Apo(a) higher than the average before ovulation induction was 1.435, 1.233, 0.678 times of those with lower than the average ( P<0.05). The risk of non-clinical pregnancy for those with LPO, TOS, and Apo(a) higher than the average on HCG day was 1.443, 1.689, 0.762 times of those with lower than average ( P<0.05). After receiver operating characteristic (ROC) curve analysis, the area under the curve(AUC) of all indicators before ovulation induction combined to predict clinical pregnancy was 0.844. The AUC of all indicators on HCG day combined to predict clinical pregnancy was 0.894. Conclusions:Peripheral blood LPO, TOS, Apo(a) levels are closely related to the number of high-quality embryos, and are the main influencing factors of pregnancy outcome. Therefore, dynamic monitoring of the above-mentioned index levels can provide a reference for the clinical improvement of the treatment plan.
4.Effect and mechanism of cathepsin S on proliferation, migration and invasion of osteosarcoma cells
Hairu Ji ; Lingwei Kong ; Sheng Cao ; Jiaxing Lü ; ; Jiaxin Li ; Chunyu Liu ; Yu Jin
Acta Universitatis Medicinalis Anhui 2022;57(9):1459-1465
Objective:
To investigate the effects of cathepsin S(Cat S) on proliferation, migration and invasion of osteosarcoma cells and its potential regulatory mechanism.
Methods:
Normal osteoblasts(hFOB) and osteosarcoma cells(SAOS2 and MG63) were selected as the subjects of this study.Cat S small interfering(si) RNA(si-Cat S) and negative control sequence(si-NC) were transfected into SAOS2 and MG63 cells to modulate the expression of Cat S in osteosarcoma cells.The experimental cells were randomly divided into four groups: hFOB group(hFOB cells),Control group(untransfected SAOS2 or MG63 cells),si-NC group(SAOS2 or MG63 cells transfected with si-NC) and si-Cat S group(SAOS2 or MG63 cells transfected with si-Cat S).The expression of Cat S in SAOS2 and MG63 cells was detected by RT-PCR and Western blot.Effect of Cat S knockdown on the proliferation of SAOS2 and MG63 cells was assessed by CCK-8 and clone formation assays.And effects of Cat S knockdown on the migration and invasion of SAOS2 and MG63 cells were determined by wound-healing and Transwell assays, respectively.Western blot assay was performed to measure the effects of SAOS2 knockdown on the expressions of apoptosis-related proteins(Bcl-2,Bax and Caspase-3) and Wnt/β-catenin pathway related proteins(LRP5,β-catenin, C-myc and Cyclin D1) in SAOS2 cells.
Results:
Compared with hFOB group, the expression of Cat S in SAOS2 group and MG63 group was upregulated(P<0.001).In addition, compared with si-NC group, the proliferation, migration and invasion of cells in si-Cat S group were reduced(P<0. 001). Results of Western blot showed that compared with si-NC group,the expression of Bcl-2 in si-Cat S group was downregulated,while the expression of Bax and Caspase-3 were upregulated(P<0. 001). Meanwhile,compared with si-NC group,the expression of LRP5,β-catenin,C-myc and Cyclin D1 in si-Cat S group was downregulated(P<0. 001).
Conclusion
Cat S siRNA knockdown can inhibit the proliferation,migration and invasion of osteosarcoma cells and induce apoptosis by regulating Wnt/β-catenin pathway,indicating that Cat S may be one of the potential targets for the treatment of osteosarcoma.