1.Protective effects of propofol on focal ischemia-reperfusion injury in rats
Yingmin CAI ; Haitao HU ; Meina WANG ; Xiaoya MA
Chinese Journal of Tissue Engineering Research 2006;10(4):175-177
BACKGROUND: In clinical, propofol can contract cerebral vessels, decrease cerebral blood flow, decrease brain metabolic oxygen consumption,which can decrease pressure in brain. Studies prove that propofol can protect endothelial cell that may be injuried by active oxygen injury and also decrease nerves injury of experimental rats with cerebral ischemia.OBJECTIVE: To investigate the protective effects of propofol on cerebral ischemia-reperfusion injury in rat and its mechanism.DESIGN: Randomized and controlled study.SETTING:Anesthesiological Department of the Second Affiliated Hospital of Xi'an Jiaotong University.PARTICIPANTS: The experiment was conducted at Pharmacological Laboratory of Medical College of Xi' an Jiaotong University in 2004. Totally 40 healthy male SD rats, aged 3-4 months, weighting 200-300 g, were divided randomly into four groups: Model group, control group, nimodipine group and propofol group, with 10 in each group.METHODS: The rats were anesthetized by intraperitoneal methods with ketamine and propofol separately. When righting reflex was abolished, external carotid artery was separated and ligated. A nylon thread was put at the stump site of external carotid artery without ligation. Model group: 10 mL normal saline was injected into intraperitone in 10 minutes before ischemia.Control group: 10 mL normal saline was injected into intraperitone at the end of operation. Nimodipine group: 10 g/L nimodipine (1 mg/kg) was injected into intraperitone in10 minutes before ischemia. Propofol group: 10 g/L propofol (110 mg/Kg) was injected into intraperitone in 10 minutes before ischemia. When ischemia was lasted for 3 hours, nylon thread was with drawed for reperfusion. When reperfusion was lasted for 3 hours, blood samples were obtained from orbit. Skulls were opened and brains were removed.Effect of propofol on cerebral ischemia-reperfusion injury was observed.MAIN OUTCOME MEASURES: Infarction area, cerebral water content,serum lactate dehydrogenase (LDH) and creatine kinase (CK) levels, brain superoxide dismutase (SOD) activity, malondialdehyde (MDA) and Ca2+levels were measured. Ultrastructure of brain tissue was examined under electron microscope.RESULTS: ①Infarct area in propofol group was significantly smaller than that in model group [(10.45±3.65, 19.68±4.03)%, (t=3.493,P < 0.01)]. ② CK level was lower in propofol group than that in model group [(471±200,1 930±917) IU/L, (t=3.493, P < 0.01)]; and LDH level in propofol group [(8 240±2 580) U/L] was significantly different from that in model group [(15 470±2 680) U/L, (t=3.441, P < 0.01)]; And water content in brain tissue was lower in propofol group than that in model group [(78.2±2.4,82.9±2.9)%, (t=3.321, P < 0.01)]. ③ The death rate of rats was 13.6%in propofol group, and 47.6% in model group, the former was decreased obviously as compared with the latter, and the difference was significant (t=6.21,P < 0.05). ④ SOD activity was (1 690±780) U/g in propofol group and (830±110) U/g in model group, the difference was significant (t=3.420, P < 0.01); but MDA content was obviously lower in propofol group than that in model group [(0.058±0.014, 0.115±0.047) μmol/g, (t=3.336, P < 0.01)].CONCLUSION: Propofol has protective effect on cerebral ischemia-reper fusion injury in rats, and the mechanism is related with inhibition of Ca2+overloading and lipid peroxidation.
2.Correlation of reversely increased level of plasma glucose during pregnancy to the pregnancy outcome
Xiaoya SHEN ; Shilei PAN ; Wei CAI ; Baoping ZHU
Medical Journal of Chinese People's Liberation Army 2017;42(1):47-51
Objective To explore the correlation of the reversely increased results of 75g oral glucose tolerance test (OGTT) during pregnancy to the pregnancy outcome, so as to provide a reliable theoretical basis of the early intervention for the pregnant women with high plasma glucose.Methods The clinical data of 461 cases were retrospectively analyzed. Patients were chosen from the pregnant women undergoing routine antenatal examination in our hospital during 2014. According to the results of 75g OGTT, 226 patients were analyzed as the observation group, in whom the level of postprandial 2-hour plasma glucose was higher than that of postprandial 1-hour plasma glucose. Meanwhile 235 pregnant women with or without gestational diabetes mellitus (GDM) were randomly selected as the control group.Results The levels of fasting plasma glucose and 1-hour postprandial plasma glucose were lower, but those of 2-hour postprandial plasma glucose was higher in observation group than in control group (P<0.01). No statistical difference existed between the two groups (P>0.05) in the incidences of polyhydramnios, oligohydramnios, fetal growth restriction (FGR), premature labor (PTL), pregnancy induced hypertension (PIH), complicated with premature rupture of membrane (PROM), intrauterine fetal death (IUFD) and non scar uterus cesarean section rate (CSR). Compared with the observation group, the rates of neonatal dysplasia and neonatal asphyxia and the newborn transfer rate were lower in the control group, of which the newborn transfer rate was statistically different (P<0.01).Conclusions There might be a delayed plasma glucose metabolism in the patients with reversely increased result of 75g OGTT during pregnancy, which may affect the long-term prognosis of the newborn. Therefore, more attention should be paid to such patients with reversely increased result of 75g OGTT.
3.Revision and reliability and validity testing of the Arteriovenous Fistula Assessment Scale
Jinhong MIAO ; Ruoyu CAI ; Yutian LEI ; Xiaoxing WANG ; Kankan WANG ; Xiaoya ZHANG
Chinese Journal of Practical Nursing 2023;39(27):2127-2132
Objective:To translate and revise the Arteriovenous Fistula Assessment Scale (AVF-AS), test the reliability and validity of the Chinese version of AVF-AS.Methods:The modified Brislin translation model was used to translate, back translate and cross culture adjust AVF-AS, forming the Chinese version of AVF-AS. Using the convenient sampling method, 220 hemodialysis patients from the First Affiliated Hospital of Zhengzhou University were selected for investigation from July to September 2022. Two weeks later, 30 patients were randomly selected for retesting. The valid data were used for project analysis and reliability and validity evaluation.Results:The Chinese version of AVF-AS consisted of 3 factors and 18 items. Consistency level between evaluators was 0.94, the item level content validity index was 0.83-1.00, average scale level content validity index was 0.94, and the calibration validity was 0.68.Three common factors(autogenous arteriovenous fistula blood flow, stenosis and ischemia, puncture location) were extracted from exploratory factors, and the cumulative variance contribution rate was 75.255%. The total scale's Cronbach α was 0.946, the half reliability of each dimension was 0.826 - 0.898, and the test-retest reliability was 0.907.Conclusions:The Chinese version of AVF-AS has good reliability and validity, and can be used as an effective tool to evaluate the autogenous arteriovenous fistula functional status of hemodialysis patients in China.
4.Relaxant Effect and Underlying Mechanisms of Evodiamine on Isolated Myometrium of Rats
Xiaoya LI ; Jinyu LIU ; Yuxue MU ; Lin PENG ; Shasha GE ; Xin ZHAO ; Yulin LIN ; Dayong CAI
China Pharmacist 2017;20(10):1713-1717
Objective: To study the relaxant effect and underlying mechanisms of evodiamine on isolated myometrium of rats. Methods:Prostaglandin F2α( PGF2α) was used to induce isolated myometrium contraction. The relaxant effect of evodiamine and the influence of capsazepine (an antagonist of transient receptor potential cation channel, subfamily V, member 1, TRPV1), U73122 (an antagonist of phospholipase Cβ,PLCβ) and W-7 ( an antagonist of camodulin, CaM) on the relaxant effect of evodiamine on myometri-um were observed respectively by biological function experiments. The median effective concentration ( EC50 ) was analyzed by non-line-ar various slope regressions using Prism-5. 01 software. Results:Evodiamine showed concentration-dependent relaxant effect on PGF2α-induced myometrium contraction with the EC50of 9.56 ×10 -9mol·L-1. Incubation with capsazepine (6.30 ×10 -11 mol·L-1), U73122 (2. 57 × 10 -11 mol·L-1 ) and W-7 (5. 65 × 10 -13 mol·L-1 ) markedly increased the relaxant effect of evodiamine, the EC50 of evodiamine decreased and dose-effect curves left shifted. The order of EC50 was as follows: W-7- evodiamine (8. 88 × 10 -15 mol· L-1) < capsazepine-evodiamine (7.35 ×10 -13 mol·L-1) < U73122-evodiamine (1.95 ×10 -12mol·L-1). Conclusion: Evodia-mine can inhibit myometrium contraction induced by PGF2αobviously, and the mechanisms are probably related to TRPV1, PLCβand CaM.
5.Relaxant Effect and Underlying Mechanisms of Evodiamine on Isolated Myometrium of Rats
Xiaoya LI ; Jinyu LIU ; Yuxue MU ; Lin PENG ; Shasha GE ; Xin ZHAO ; Yulin LIN ; Dayong CAI
China Pharmacist 2017;20(10):1713-1717
Objective: To study the relaxant effect and underlying mechanisms of evodiamine on isolated myometrium of rats. Methods:Prostaglandin F2α( PGF2α) was used to induce isolated myometrium contraction. The relaxant effect of evodiamine and the influence of capsazepine (an antagonist of transient receptor potential cation channel, subfamily V, member 1, TRPV1), U73122 (an antagonist of phospholipase Cβ,PLCβ) and W-7 ( an antagonist of camodulin, CaM) on the relaxant effect of evodiamine on myometri-um were observed respectively by biological function experiments. The median effective concentration ( EC50 ) was analyzed by non-line-ar various slope regressions using Prism-5. 01 software. Results:Evodiamine showed concentration-dependent relaxant effect on PGF2α-induced myometrium contraction with the EC50of 9.56 ×10 -9mol·L-1. Incubation with capsazepine (6.30 ×10 -11 mol·L-1), U73122 (2. 57 × 10 -11 mol·L-1 ) and W-7 (5. 65 × 10 -13 mol·L-1 ) markedly increased the relaxant effect of evodiamine, the EC50 of evodiamine decreased and dose-effect curves left shifted. The order of EC50 was as follows: W-7- evodiamine (8. 88 × 10 -15 mol· L-1) < capsazepine-evodiamine (7.35 ×10 -13 mol·L-1) < U73122-evodiamine (1.95 ×10 -12mol·L-1). Conclusion: Evodia-mine can inhibit myometrium contraction induced by PGF2αobviously, and the mechanisms are probably related to TRPV1, PLCβand CaM.
6.Dose-effect Relationship of Xianfu Ointment and its Decomposed Recipes on Chronic Eczema
Lin PENG ; Yuxue MU ; Jinyu LIU ; Xiaoya LI ; Shasha GE ; Shuang LI ; Xin ZHAO ; Yulin LIN ; Dayong CAI ; Liping SUN ; Binghua TANG ; Lianqi LIU
China Pharmacist 2018;21(5):817-823
Objective:To investigate the dose-effect relationship of Xianfu ointment and its decomposed recipes the 1-chloro-2,4-dini-trochlorobenzene(DNCB) induced chronic eczema in mice, and confirm the median effective dose (ED50) of each formula and the synergetic effect by compatibility. Methods:DNCB was used to induce chronic eczema in C57 mice. The mice were treated with gradient dosages of the Xianfu ointment (11.71-11 662.50 mg?kg-1?d-1,k = 0.316), Anemone flaccid (0.53-530.12 mg?kg-1?d-1,k = 0.316), Xianfu ointment without Anemone flaccid (11.18-11 132.40 mg?kg-1?d-1,k =0.316),respectively. The pathological features were observed after hematoxylin-eosin staining. The volume ratio of epidermides and the number of lymphocyte infiltrated in dermis were analyzed with morphometry. The serum levels of IL-2,IFN-γ,IL-4,and IL-13 were detected by ELISA assay. The ED50was calculated by non-linear regression with various slope using Prism-5.0 software.Results:The effects of Xianfu ointment and its decomposed recipes on chronic eczema showed a dose-dependent tendency. The dose-response curves showed"S"shape. The efficacy of Xianfu ointment on chronic eczema was the most significant among the three formulas, which was demonstrated by decreased epidemical thicknes (ED50= 377.90 mg?kg-1?d-1), reduced infiltrated lymphocyte number(ED50= 153.20 mg?kg-1?d-1), increased serum IL-2(ED50=608.90 mg?kg-1?d-1) and IFN-γ (ED50= 205.50 mg?kg-1?d-1) levels, and decreased serum IL-4(ED50= 198.70 mg?kg-1?d-1) and IL-13 levels (ED50= 117.60 mg?kg-1?d-1). And the dose-effect curves of Anemone flaccid and Xianfu ointment without Anemone flaccid groups were both right shift when compared with that of Xianfu ointment. Conclusion:Xianfu ointment and its decomposed recipes can effectively treat chronic eczema. Anemone flaccid has obvious compatibility synergy in the whole formula. The effects of Xianfu ointment is most significant.
7.Dose-effect Relationship of Yuning Ointment and its Decomposed Recipes on Acne in Mice
Yuxue MU ; Lin PENG ; Xiaoya LI ; Jinyu LIU ; Shasha GE ; Shuang LI ; Xin ZHAO ; Lianqi LIU ; Binghua TANG ; Dayong CAI ; Liping SUN
China Pharmacist 2018;21(6):949-955
Objective: To study the dose-effect relationship of Yuning ointment and its decomposed recipes in the treatment of oleic acid induced acne in mice. Methods: Oleic acid was administrated to the back (2 cm ×2 cm) of the mice (once a day) for 21 days to induce acne. At d22, the gradient dosage of Anemone flaccida crude drug (1. 06-1 060. 23 mg?kg-1?d-1,k=3. 16), Yuning oint-ment without Anemone flaccida crude drug (4. 73-1 767. 75 mg?kg-1?d-1, k=3. 16) and Yuning ointment (2. 84-2 827. 28 mg?kg-1?d-1, k=3. 16) was respectively administrated to the back of mice for 14 days. The pathological changes of skin were observed by hematoxylin-eosin (HE) staining. The diameter of sebaceous glands and the ratio of follicular keratinization area were morphomet-rically analyzed. The serum levels of IL-1, IL-6 and TNF-α were detected by ELISA assay. The median effective dosages (ED50) of A-nemone flaccida in the three prescriptions were regressed by Prism 5. 01 software to determine the prescription dose-effect. Results: All the therapy groups were with significantly relieved pathological changes of sebaceous glands hypertrophy and follicular keratinization, and decreased serum levels of IL-1, IL-6 and TNF-α in a dose-dependent manner. The dose-response curves showed an "S" shape. A-mong the three therapy groups, the effect of Yuning ointment was the best. The ED50of Yuning ointment regressed by Anemone flaccida dose was 0. 28-fold for improving sebaceous glands hypertrophy, 0. 14-fold for inhibiting follicular keratinization, and 0. 15-, 0. 49-and 0. 24-fold for decreasing serum levels of IL-1, IL-6 and TNF-α. . Regressed by Yuning ointment without Anemone flaccida, the ED50of Yuning ointment was lower than Yuning ointment without Anemone flaccid in terms of improving pathological changes and inhibiting the secretion of cytokines. Conclusion: Yuning ointment can prevent and treat acne through regulating immune function. And the prescrip-tion compatibility can enhance the effects of Anemone flaccida.
8.Effect of GSK-3β-mediated DRP1 on inhibition of primary hippocampal neuronal growth induced by aluminum
Meng LI ; Liyuan LU ; Xiaoyu HE ; Changxin XIANG ; Xiaoya CAI ; Huifang ZHANG
Journal of Environmental and Occupational Medicine 2022;39(10):1095-1101
Background Aluminum (Al) can cause irreversible damage to neurons and synapses function, and the mechanism may be connected to mitochondrial damage caused by glycogen synthase kinase-3β (GSK-3β) regulating dynamin-related protein 1 (DRP1), resulting in inhibition of the growth of neuronal protrusions. Objective To investigate the role of GSK-3β regulating DRP1 in the inhibition of primary hippocampal neurite growth induced by Al. Methods Neurons were extracted from the hippocampus of newborn mice (≤24 h old) for primary culture. On day 6, the purity of neurons was detected by immunofluorescence. On day 10, neurons with good growth state were selected for Al exposure and GSK-3β inhibitor SB216763 (SB) intervention. The experiment design included a blank control group, a dimethyl sulfoxide (DMSO) group, an Al (20 μmol·L−1) group, a SB (1 μmol·L−1) group, and a SB (1 μmol·L−1) + Al (20 μmol·L−1) group. After primary hippocampal neurons were treated with Al or SB for 48 h, cell viability was detected by CCK-8 assay, the mitochondrial morphology of primary hippocampal neurons was observed by transmission electron microscopy, the total protrusion length of primary hippocampal neurons was scanned and analyzed by laser confocal imaging, and their complexity was analyzed by Sholl analysis. The expression levels of phospho-GSK-3β, GSK-3β, and DRP1 were detected by Western blotting. Results The immunofluorescent results showed that the purity of primary neurons was higher than 90%. After the Al exposure and the SB intervention for 48 h, compared with the blank control group, there was no obvious difference in cell viability in the DMSO group and the SB group (P>0.05), and the Al group showed reduced cell viability (P=0.006); there was no obvious difference in cell viability between the SB+Al group and the Al group (P>0.05). Compared with the blank control group, there was no obvious difference in the average total length of protrusion in the DMSO group and the SB group (P>0.05), and the Al group showed reduced average total length of neurite (P<0.001); the average total neurite length in the SB+Al group was significantly increased compared with that in the Al group (P=0.001). The results of Sholl analysis revealed that, within 130 μm from the cytosol, the number of intersections of neurons in each group increased with the increase of distance. Above 130 μm from the cytosol, the number of intersections of neurons in each group decreased gradually with increase of distance. At 130 μm and 310 μm from the cytosol, compared with the blank control group, the number of neuronal intersections in the DMSO group and the SB group had no obvious difference (P>0.05), and that in the Al group was significantly reduced (P<0.05); there was no obvious difference in the number of neuronal intersections between the SB+Al group and the Al group (P>0.05). The mitochondrial structure of the blank control group was complete and the crest was clearly visible; there was no apparent variation in the mitochondrial structure in the DMSO group and the SB group; the mitochondria in the Al group were vacuolated and the crista disappeared; the SB+Al group showed clearer crista than the Al group. The difference in GSK-3β phosphorylation level among groups was statistically significant (F=45.841, P<0.001). Compared with the blank control group, the GSK-3β phosphorylation level showed not significantly different in the DMSO group (P>0.05), increased in the SB group (P=0.022), and significantly reduced in the Al group (P<0.001); the GSK-3β phosphorylation level was significantly higher in the SB+Al group than in the Al group (P<0.001). The difference in DRP1 protein level among groups was statistically significant (F=8.389, P=0.003). Compared with the blank control group, the DRP1 protein levels in the DMSO group and the SB group were not significantly different (P>0.05), and significantly increased in the Al group (P=0.001); the DRP1 protein level in the SB+Al group was significantly lower than that in the Al group (P=0.029). Conclusion Al may increase the level of DRP1 protein by activating GSK-3β, causing mitochondrial damage and inhibiting neuronal protrusions growth.
9.Effect of aluminum-maltolate on primary hippocampal neuron neurite damage by regulating CRMP2 through GSK-3β in mice
Huifang ZHANG ; Yingchao HAN ; Xiaoya CAI ; Changxin XIANG ; Qiao NIU
Journal of Environmental and Occupational Medicine 2021;38(11):1207-1213
Background Aluminum can induce irreversible structural and synaptic functional damage, and the associated mechanism may be related to the neurite damage regulated by glycogen synthase kinase-3β (GSK-3β)/collapsin response mediator protein 2 (CRMP2). Objective This experiment is conducted to investigate the effect of aluminum-maltolate [Al(mal)3] on primary hippocampal neuron neurites in mice, and reveal the role of GSK-3β-CRMP2 in this process. Methods The hippocampus of newborn ICR mice (≤ 24 h old) was used for primary neuronal cultures. On the 5th day in vitro (DIV5), neuron purity detection were performed by confocal laser scanning microscopy. On DIV7, the neurons were transfected with lentiviral vector-mediated mNeonGreen. On DIV10, the neurons with mNeonGreen fluorescence in good growth state were treated with Al(mal)3. The stage I experimental groups were blank control group, maltol group, 10 µmol·L−1 Al group, 20 µmol·L−1 Al group, and 40 µmol·L−1 Al group. Then 20 µmol·L−1 Al was used to establish a model of neurite injury and for the intervention. The stage II experimental groups were blank control group, dimethyl sulfoxide (DMSO) group, Al (20 µmol·L−1) group, SB (GSK-3β inhibitor, 1 µmol·L−1), and SB (1 µmol·L−1)+Al (20 µmol·L−1) group. CCK-8 method was used to detect the viability of neurons. The primary hippocampal neurons of mice were scanned with high content analysis system at 0 h and 48 h after Al or SB treatment, and the density and length of neurites were analyzed. Western blotting was used to detect the expression and phosphorylation levels of CRMP2 and GSK-3β in primary hippocampal neurons of mice. Results The immunofluorescence results showed that the purity of primary neurons was more than 90%. Compared with the blank control group in stage I, the cell viability rates of the 10, 20, and 40 µmol·L−1 Al groups were decreased after 48h of Al(mal)3 treatment (P<0.05), while the cell viability rate of the maltol group had no significant change. There was no significant difference in cell viability rate among the DMSO group, the SB group, and the control group after 48h of SB treatment, and the viability rate of neurons in the SB+Al group was higher than that in the Al group (P<0.05) in stage II. The 48 h/0 h ratios of average number and length of neurites in the control group were 90.13%±11.70% and 113.24%±8.34%, respectively. The 48 h/0 h ratios in the Al group were 56.47%±16.36% and 62.06%±6.75%, respectively, which were lower than those in the control group (P<0.05). The 48 h/0 h ratios of average number of neurites in the SB group (99.03%±21.83%) was not significantly different from that in the control group, but the 48 h/0 h ratio of average length of neurites in the SB group (128.72%±15.39%) was higher than that in the control group (P<0.05). The 48 h/0 h ratios of average number (72.59%±10.89%) and length of neurites (93.84%±14.65%) in the SB+Al group were significantly increased compared with those in the Al group (P<0.05). Western blotting results showed that: There was no significant difference in GSK-3β protein level among all groups; compared with the control group (1.00±0.18), the protein level of p-GSK-3β in the Al group (0.45±0.05) was significantly decreased, and that in the SB group (1.32±0.23) was significantly increased; the protein level of p-GSK-3β in the SB+Al group (0.80±0.05) was significantly higher than that in the Al group (P<0.05). Compared with the control group (1.00±0.07), the CRMP2 protein level in the Al group (0.66±0.11) was significantly decreased (P<0.05), while that in the SB group (1.01±0.02) was not significantly changed. Compared with the control group (1.00±0.13), the p-CRMP2 protein level in the Al group (1.50±2.18) was significantly increased, and that in the SB group (0.62±0.09) was significantly decreased (P<0.05); the protein level of p-CRMP2 in the SB+Al group (1.28±0.24) was lower than that in the Al group (P<0.05). Conclusion Aluminum may activate GSK-3β, increase CRMP2 phosphorylation level, and damage neurite growth.