Animal Feed ; Animals ; Ascorbic Acid ; pharmacology ; Cadmium ; analysis ; DNA ; analysis ; Hepatocytes ; chemistry ; Rats ; Rats, Wistar ; Selenium ; analysis

OBJECTIVE:To establish a method for simultaneous determination of paeoniflorin,berberine hydrochloride and ammonium glycyrrhizinate in Xiaoer huadu powder.METHODS:HPLC method was adopted.The separation was performed on Waters SunFireTM-C18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 238 nm,and column temperature was 25 ℃.The sample size was 10 μL.RESULTS:The linear ranges of paeoniflorin,berberine hydrochloride and ammonium glycyrrhizinate were 8.808-88.08 μg/mL(r=0.999 8),1.778-17.78 μg/mL(r=0.999 6),2.533-25.33 μg/mL(r=0.999 9),respectively.LOQ were 4.404,0.889,2.533 μg/mL;LOD were 1.101,0.445,1.267 μg/mL.RSDs of precision,stability and reproducibility tests were all lower than 2％.The recoveries were 95.08％-99.61％ (RSD=1.77％,n =9),96.93％-99.94％ (RSD=0.92％,n=9),98.33％-102.05％ (RSD=1.27％,n=9).CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for simultaneous determination of paeoniflorin,berberine hydrochloride and ammonium glycyrrhizinate in Xiaoer huadu powder.

Objective To study the effects of morroniside on the expression of Wnt signaling-related transcription factors neurogenin 2 (Ngn2), Pax6 and Tbr2 in the ischemic ipsilateral cortex 7 days after cerebral ischemia-reperfusion in rats. Methods 15 male Sprague-Dawley rats were randomly divided into sham group (n=3), ischemia group (n=3), and morroniside groups (low, medium and high dosage groups, n=3). The middle cerebral artery were occluded for 30 min, and re-perfused. Morroniside was administered intragastrically once a day at dose of 30 mg/kg, 90 mg/kg and 270 mg/kg 3 hours after operation. The expression of Ngn2, Pax6 and Tbr2 in the ischemic ipsilateral cortex were detected with Western blotting analysis 7 days after operation. Results The expression of Ngn2 increased in the ischemia group compared with the sham group (P<0.05), and it further increased the morroniside groups of medium and high dosage compared with the ischemia group (P<0.01). There was no significant difference between the ischemia group and sham group in the expression of Pax6, while it increased the morroniside groups of medium and high dosage compared with the ischemia group (P<0.01). There was no significant difference among all the groups in the expression of Tbr2. Conclusion Morroniside could increase the expression of Ngn2 and Pax6 in the ischemic ipsilateral cortex 7 days after ischemia-reperfusion in rats, suggesting promoting the neurogenesis after ischemia.

Objective To study the effects of morroniside on the expression of matrix metalloproteinase (MMP) -2 and MMP-9 in the peri- infarct cortex 3 days after cerebral ischemia- reperfusion. Methods 15 male Sprague-Dawley rats were randomly divided into sham group (n=3), ischemia group (n=3), and morroniside groups (low, medium and high dosage groups, n=3). The middle cerebral artery were occluded for 30 min, and re-perfused. Morroniside was administered intragastrically once a day at dose of 30 mg/kg, 90 mg/kg and 270 mg/kg 3 hours after operation. The expression of MMP-2 and MMP-9 in peri-infarct cortex were detected with immunohistochemistry staining 3 days after operation. Results The expression of MMP-2 and MMP-9 increased in the ischemia group compared with the sham group (P<0.01), and it decreased in all the morroniside groups compared with the ischemia group (P<0.01). Conclusion Morroniside could decrease the expression of MMP-2 and MMP-9 in the peri-infarct cortex 3 days after ischemia, suggesting protecting the function of blood-brain barrier from ischemia.

Objective To investigate the effects of Morroniside on H2O2-induced oxidative injury in SH-SY5Y neuroblastoma cells.MethodsSH-SY5Y cells were pre-incubated with Morroniside(1,10,and 100 μmol/L)for 24 h prior to exposure to H2O2(500 μmol/L)for 18 h.The content of reactive oxygen species(ROS),nitric oxide(NO),glutathione(GSH)were determined.ResultsPretreatment the cells with Morroniside(10 and 100 μmol/L)lowered H2O2-induced ROS release by 37%(P<0.01)and 27%(P<0.05)versus H2O2,and NO release in a dose-dependent manner by 31%(P<0.05),53%(P<0.01)versus H2O2.Morroniside(1,10,and 100 μmol/L)increased GSH level in cells treated with H2O2 in comparison with cells exposed only to H2O2,respectively.ConclusionMorroniside shows neuroprotection effect against H2O2-induced oxidation injury in SH-SY5Y cell.

Neural stem cells are a category of cells with self-renewal and differentiation potential. Proliferation and differentiation of neural stem cells have closely relation with growth factor, cell factor, environmental signal, and so on. In recent years, evidence indicates that neural stem cells possessing multipotency naturally occur in adult brain. The found of adult neural stem cells have changed the theory that central nervous system cannot regenerative. Chinese Traditional Medicine can effectively induce adult neural stem cells to proliferate and differentiate, opening up unprecedented direction in the research for the treatment of nerve regeneration, and showing great potency for curing cerebral ischemic injury.

Brain ; pathology ; Cerebrovascular Disorders ; diagnosis ; Child ; Diagnosis, Differential ; Female ; Humans ; Magnetic Resonance Imaging ; Neurofibromatosis 1 ; diagnosis ; therapy ; Treatment Outcome

This study aimed to investigate the inhibitory effect of tubuloside B (Tub B) on amyloid β-protein (Aβ), and analyse the potential mechanism. A model of amyloid fibril was established by incubation of Aβ_1-42 in vitro. Thioflavin-T (ThT), Congo red (CR), 8-anilino-1-naphthene sulfonic acid (ANS) staining and transmission electron microscopy (TEM) were applied to detect the suppression of Tub B on the formation of Aβ_1-42 fibril. Circular dichroism (CD) was used to analyse the regulatory effect of Tub B on the secondary structure of Aβ_1-42. 3-(4,5-Dimethyl-2-thiazole) -2,5-diphenyltetrazolium bromide (MTT) and red blood cell hemolysis experiments were used to investigate the attenuation of Tub B on Aβ_1-42 induced cytotoxicity. 2',7'-Dichlorofluorescin diacetate (DCFH-DA) staining was used to assess the expression of intracellular reactive oxygen species (ROS) induced by Aβ_1-42. And molecular docking experiment was used to explore the interaction between Tub B and Aβ_1-42. The results indicated that Tub B could inhibit Aβ_1-42 fibrillization in a certain extent, which retarded the structural transition of α-helix to β-sheet of Aβ_1-42, hampered the exposure of hydrophobic regions, and attenuated amyloid-induced cytotoxicity and hemolysis. In summary, Tub B can prevent the formation of Aβ_1-42 amyloid fibril, which may be related to its antioxidant activity and hydrogen bonding and hydrophobic interactions with protein molecules. All animal experiments were approved by the Experimental Animal Research Center of Air Force Medical University (No. 20190051).

ObjectiveTo investigate the effects of morroniside on super oxide dismutase (SOD) and neurons in rats cortex with focal cerebral ischemia/reperfusion. MethodsThe animal model was induced by middle cerebral artery occlusion with suture embolus, cerebral ischemia 30 min and reperfusion 3 d or 7 d. Vitamin E for the positive control. The content of SOD was detected with spectrophotometry and the nerve cells was observed with immunohistochemistry. ResultsCompared with model group, morroniside （270 mg/kg）increased the activity of SOD and the number of neurons （30 mg/kg, 90 mg/kg, 270 mg/kg） significantly. ConclusionMorroniside may have neuroprotective effect and increasing the activity of SOD in rats cortex.

Objective To investigate the effects of morroniside on total antioxidative capacity (T-AOC) in rat cortex with focal cerebral ischemia/reperfusion. Methods The animal model was induced with occlusion of middle cerebral artery. The T-AOC was detected with spectrophotometer. Results Compared with model group, morroniside （270 mg/kg） increased the T-AOC significantly. Conclusion Morroniside may take the neuroprotection through increased T-AOC of rat cortex.