ABSTRACT
OBJECTIVE@#To study the protective effect of forsythiaside B (FB) against cerebral oxidative stress injury induced by cerebral ischemia/reperfusion (I/R) in mice and explore the underlying mechanism.@*METHODS@#Ninety C57BL/6 mice were randomized into sham-operated group, middle cerebral artery occlusion (MCAO) model group, and low-, medium and highdose (10, 20, and 40 mg/kg, respectively) FB groups. The expression levels of MDA, ROS, PCO, 8-OHdG, SOD, GSTα4, CAT and GPx in the brain tissue of the mice were detected using commercial kits, and those of AMPK, P-AMPK, DAF-16, FOXO3 and P-FOXO3 were detected with Western blotting. Compound C (CC), an AMPK inhibitor, was used to verify the role of the AMPK pathway in mediating the therapeutic effect of FB. In another 36 C57BL/6 mice randomized into 4 sham-operated group, MCAO model group, FB (40 mg/kg) treatment group, FB+CC (10 mg/kg) treatment group, TTC staining was used to examine the volume of cerebral infarcts, and the levels of ROS and SOD in the brain were detected; the changes in the protein expressions of AMPK, P-AMPK, DAF-16, FOXO3 and P-FOXO3 in the brain tissue were detected using Western blotting.@*RESULTS@#In mice with cerebral IR injury, treatment with FB significantly reduced the levels of ROS, MDA, PCO and 8-OHdG, increased the activities of antioxidant enzymes SOD, GSTα4, CAT and GPx, and enhanced phosphorylation of AMPK and FOXO3 and DAF-16 protein expression in the brain tissue (P < 0.01). Compared with FB treatment alone, the combined treatment with FB and CC significantly reduced phosphorylation of AMPK and FOXO3, lowered expression of DAF-16 and SOD activity, and increased cerebral infarction volume and ROS level in the brain tissue of the mice (P < 0.01).@*CONCLUSION@#FB inhibits oxidative stress injury caused by cerebral I/R in mice possibly by enhancing AMPK phosphorylation, promoting the downstream DAF-16 protein expression and FOXO3 phosphorylation, increasing the expression of antioxidant enzymes, and reducing ROS level in the brain tissue.
Subject(s)
Mice , Animals , AMP-Activated Protein Kinases/metabolism , Antioxidants/metabolism , Reactive Oxygen Species , Mice, Inbred C57BL , Brain Ischemia , Oxidative Stress , Infarction, Middle Cerebral Artery , Reperfusion Injury , Reperfusion , Superoxide Dismutase/metabolismABSTRACT
AIM: To study the effects of forsythiaside B on HeLa cells proliferation by inhibiting NF-κB through p21/Cyclin E/CDK2 signaling pathway. METHODS: The cell activity of HeLa cells was observed by MTT assay after 24 h, 48 h and 72 h treatment with different concentration (1, 2, 4 μmol/L) of forsythiaside B. The clone formation of HeLa cells was evaluated with the condition of different concentrations of forsythiaside B. The expression of transcription factor NF-κB (p-p65 and p65) was determined, and the expression of p21, Cyclin E and CDK2 were detected as well.RESULTS:Forsythioside B inhibited the clone formation of HeLa cells in concentration dependent manner, and the inhibitory effect on HeLa cells' proliferation by forsythioside B was in both time and concentration dependent manner. Forsythioside B up-regulated p21 protein expression and down-regulated p-p65, Cyclin E and CDK2 protein expression. CONCLUSION:Forsythioside B inhibits the transcription factor NF-κB and affects the p21/Cyclin E/CDK2 signaling pathway, thus inhibiting the proliferation of HeLa cells.
ABSTRACT
OBJECTIVE:To establish a method for the simultaneous determination of the contents of Shuanghuanglian freeze-dried powder in placental perfusate. METHODS:HPLC was performed on the column of Agilent Zorbax-C18 with mobile phase of acetonitrile-1%formic acid aqueous solution(gradient elution)at flow rate of 1.0 ml/min,the internal standard was puera-rin,detection wavelength was 280 nm,column temperature was 25 ℃ and sample volume was 10 μl. RESULTS:There was a lin-ear range between linear ranges and peak area of 8 ingredients(r≥0.999 0);RSDs of within-day and inter-day precision tests were no more than 1.9%,repeatability tests was no more than 7.3%;average recoveries were in the range of 92.73%-112.37%(RSD=3.2%-8.2%,n=6);and average matrix effects were 90.33%-105.78%(RSD=3.2%-8.0%,n=6). CONCLUSIONS:The method is rapid,sensitive and specific,and can be used to the simultaneous determination of the contents of 8 ingredients of Shuan-ghuanglian freeze-dried powder in placental perfusate.
ABSTRACT
Objective: To study the technology for separation and purification of phenylethanoid glycosides from Callicarpa kwangtungensis by macroporous resin. Methods: The absorption and separation properties of 10 kinds of absorption resins were studied and the separation and purification technological process of phenylethanoid glycosides from C. kwangtungensis were optimized by HPLC with the total content of forsythiaside B and poliumoside as an index. Results: Using X-5 resin column as adsorbent, the optional conditions were as follows: loading qualities were 32.79 mg/g, the resin was washed by 4 BV distilled water to remove impurity and 7 BV 30% ethanol to elute phenylethanoid glycosides, the eluting velocity was 2 BV/h, and the content of phenylethanoid glycosides in the extract was 52.8% (n = 3). Conclusion: This technology is simple and feasible, and the process with X-5 resin is an effective method to separate and purify phenylethanoid glycosides from C. kwangtungensis.