2.Effects of Xuebijing injection on vascular endothelial cell function in rats with heat stress
Zongbao YIN ; Chao JI ; Qun XIANG ; Yu HOU ; Bin XIE
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2014;(5):360-363
ObjectiveTo investigate the mechanism of protective effect of Xuebijing injection on vascular endothelial cells in rats with heat stress.Methods Ninety Sprague-Dawley(SD) rats were randomly divided into control, model and Xuebijing injection treatment groups, 30 rats in each group. Heat stress model was reproduced by placing rats in constant temperature box at 40℃, 60% relative humidity for 1 hour, Xuebijing injection group was treated by intraperitoneal injection of Xuebijing 2.5 g/kg, while the control and model groups were treated by intraperitoneal injection of normal saline 2 mL/kg, once a day only in 1 day for both groups. After model establishment, the rectum temperature, heart rate and the mean arterial pressure(MAP) were recorded at 2, 6, 12 hours in each group. At the same time, the rat abdominal aortic blood was collected and serum was separated, the enzyme-linked immunosorbent assay(ELISA) was used to determine the aortic serum levels of lipopolysaccharide(LPS), nuclear factor-κB(NF-κB) and p53, and the prothrombin time(PT), activated partial thromboplastin time(APTT) and D-dimer of venous blood were detected by automatic blood coagulation analyzer(ACLTOP).Results Compared with those in control group, the rectum temperature, heart rate, LPS, NF-κB, p53, PT, APTT, D-dimer were significantly increased, and MAP was obviously decreased in model group(P<0.05 orP<0.01). Compared with model group, the above indexes were improved significantly in Xuebijing injection treatment group at 2 hours〔rectum temperature(℃): 38.02±0.22 vs. 39.32±0.33, heart rate(bpm): 507±14 vs. 562±35, MAP(mmHg, 1 mmHg=0.133 kPa): 98±6 vs. 87±13, LPS(ng/L): 0.65±0.03 vs. 0.82±0.05, NF-κB(ng/L): 1.10±0.04 vs. 1.33±0.05, p53(ng/L): 1.33±0.03 vs. 1.73±0.02, PT(s):15.47±1.03 vs. 20.28±2.01, APTT(s): 40.26±2.46 vs. 47.46±3.51, D-dimer(μg/L): 238.54±8.32 vs. 323.12±8.14,P<0.05 orP<0.01〕.Conclusion Xuebijing injection can correct the disorders of blood PT, APTT, D-dimer via decreasing the secretion of the levels of NF-κB, p53 from vascular endothelial cells in rats with heat stress, thus the integrity of the vascular endothelium can be protected, and LPS entering into the blood stream can be inhibited.
3. Effects of solanine on microtubules system of human breast cancer MCF-7 cells
Chinese Traditional and Herbal Drugs 2012;43(1):111-114
Objective To investigate the effects of solanine on microtubular system in MCF-7 cell line. Methods Proliferation inhibition of MCF-7 cell line was evaluated by MTT assay. Cell cycle of MCF-7 cells was analyzed and the changes of a-tubulin protein and microtubule-associated protein 2 (MAP-2) protein were detected by flow cytometry. Results The IC50 of MCF-7 cells was 22.08 μg/mL. Solanine could induce MCF-7 cells arrested in S phaseand increase the levels of a-tubulin and MAP-2 in MCF-7 cell line. Conclusion Solanine could inhibit the MCF-7 cell proliferation by increasing a-tubulin and MAP-2 expression and inducing MCF-7 cells arrested in S phase.
4. Material basis of cytotoxicity in polysaccharide from Solanum nigrum
Chinese Traditional and Herbal Drugs 2011;42(11):2275-2278
Objective: To study the material basis of cytotoxicity in polysaccharide from Solanum nigrum. Methods: Crude products of polysaccharide were isolated from S. nigrum fruit with free protein being removed by Sevage method. The crude products were decolored by 10% H2O2 and precipitated by 95% ethanol treatment. Polysaccharide-protein complex was isolated by DEAE-52 fiber column and the relative molecular weight was detected by SDS-PAGE. MTT assay was used in detecting the cytotoxicity of polysaccharide-protein complex for MCF-7 cells in vitro. Then polysaccharide-protein complex was separated by SephadexG-200 gel column and the cytotoxicity was detected by MTT method. Results: Polysaccharide-protein complex was isolated by DEAE-52 fiber column and the relative molecular wight was detected by SDS-PAGE as 3.0×10 4 and 2.5×104 for the two polysaccharide-protein complexes, It suggested that IC50 was 804.51 μg/mL by MTT. Glycoproteins A andd B were gained by Sephadex G-200 gel column from polysaccharide-protein complex. The IC50 of glycoproteins A and B were 532.96 and 613.91 μg/mL, respectively. Conclusion: Material basis of cytotoxicity in polysaccharide from S. nigrum is two kinds of glycoproteins whose relative molecular weight are 3.0×104 and 2.5×104.
5. Inhibitory mechanism of oridonin on G2/M phase arrest in SGC-7901 cell growth
Chinese Traditional and Herbal Drugs 2010;41(12):2024-2026
Objective: To investigate the effect of oridonin on G2/M phase arrest of human stomach cancer SGC-7901 cell growth in vitro and its molecular mechanism. Methods: The inhibition of oridonin on SGC-7901 cells was detected by MTT assay. Effect of oridonin on the phase distribution of SGC-7901 cell cycle was observed by flow cytometry (FCM). Western blotting was used to analyze the expression of Cdk1 and CyclinB1. Results: IC50 of oridonin on SGC-7901 cells was 15. 64 μmol/L. The effect of oridonin on SGC-7901 cells presented the increasing percentages of cells in G2/M phase as the concentration of oridonin increased. With the increase of oridonin dose, the expression of Cdk1 and CyclinB1 proteins was remarkably decreased. Conclusion: Oridonin can down-regulate the expression of Cdk1 and CyclinB1, which may be the mechanism of arresting SGC-7901 cells in G2/M phase.
6.Total flavonoids of litsea coreana decreases the cytotoxicity of oxaliplatin in TM3 Leydig cells via enhancing the function of gap junction.
Bin-Bin YU ; Xu-Hui TONG ; Shu-Ying DONG ; Yu-Chen GU ; Hao JIAO ; Jie JI ; Biao QU
National Journal of Andrology 2014;20(5):400-404
OBJECTIVETo investigate the effects of total flavonoids of Litsea Coreana (TFLC) on the gap junction (GJ) intercellular communication in TM3 testicular Leydig cells and whether TFLC can reduce the cytotoxicity of oxaliplatin (OHP) in vitro.
METHODSWe detected the effect of TFLC on the dye spread of the in vitro cultured TM3 cells by parachute assay, observed changes in the expression of connexin 43 (Cx43) total protein in the TFLC-treated TM3 cells by Western blot, and determined the effects of TFLC on the expression of Cx43 on the membrane of the TM3 cells by immunofluorescence assay and on the cytotoxicity of OHP by MTT assay.
RESULTSTFLC obviously enhanced the GJ function with the increasing of the TFLC concentration in the TM3 cells. Western blot and immunofluorescence assay confirmed that TFLC significantly enhanced the expression of Cx43 total protein and Cx43 expression on the membrane of the TM3 cells. MTT assay showed that at a high cell density (confluent with GJ formation), 20 microg/ml TFLC enhanced the GJ function of the TM3 cells and reduced the cytotoxicity of OHP (P < 0.05), while at a low density (preconfluent with no GJ formation), TFLC exhibited no effect on the cytotoxicity of OHP (P > 0.05).
CONCLUSIONTFLC increases the Cx43 expression and GJ function in normal TM3 Leydig cells, and the enhancement of GJ function reduces the cytotoxicity of OHP.
Antineoplastic Agents ; toxicity ; Cell Communication ; drug effects ; physiology ; Cell Count ; Connexin 43 ; metabolism ; Flavonoids ; pharmacology ; Gap Junctions ; drug effects ; Humans ; In Vitro Techniques ; Leydig Cells ; drug effects ; ultrastructure ; Litsea ; chemistry ; Male ; Organoplatinum Compounds ; antagonists & inhibitors ; toxicity ; Proteins ; metabolism
7.Effect of pedicle subtraction osteotomy on spino-pelvic parameters in ankylosing spondylitis patients with thoracolumbar kyphosis
Bangping QIAN ; Mingliang JI ; Yong QIU ; Zezhang ZHU ; Bin WANG ; Yang YU ; Jun JIANG
Chinese Journal of Orthopaedics 2012;32(5):398-403
ObjectiveTo explore the change of spino-pelvic parameters following pedicle subtraction osteotomy (PSO) for thoracolumbar kyphosis secondary to ankylosing spondylitis(AS).MethodsTwenty-one AS patients with thoracolumbar kyphosis,who underwent PSO at L1 level from July 2006 to October 2010 in our hospital,were retrospectively reviewed.There were 18 males and 3 females with a mean age of 35.6 years (range,21-53 years).The pre- and post-operative thoracic kyphosis(TK),lumbar lordosis (LL),globe kyphosis (GK),angle of the fusion levels (AFL),sagittal imbalance (SVA),pelvic incidence (PI),sacral slope (SS) and pelvic tilting (PT) were measured.ResultsSignificant differences were observed in terms of the improvement of LL,PT,SS,SVA,GK and AFL (P< 0.01).The alteration of LL showed significant correlation with the change of PT (r=0.59,P=0.005),SS (r=0.64,P=0.002),SVA (r=0.49,P=0.025),and AFL (r=0.60,P=0.004).The change of PT exhibited cardinal correlation with the change of SS(r=0.94,P=0.000).The improvement of AFL significantly correlated with the improvement of SS(r=0.61,P=0.003),PT (r=0.59,P=0.005).ConclusionThe change of the sagittal spino-pelvic profile following PSO in AS-related thoracolumbar kyphosis is closely related with the improvement of LL.
8.Effect of curcumin on inflammatory responses in hippocampus during global cerebral ischemia-reperfusion injury in hypertensive rats
Xiaodan WANG ; Lulu SHEN ; Chenchen YU ; Hong CAO ; Bin JI ; Jun LI
Chinese Journal of Anesthesiology 2014;34(6):753-757
Objective To evaluate the effect of curcumin on the inflammatory responses in the hippocampus during global cerebral ischemia-reperfusion (I/R) injury in hypertensive rats.Methods Forty-eight SPF male Wistar-Kyoto rats,aged 8 weeks,weighing 180-200 g,were randomly divided into 2 groups (n =24 each) using a random number table:sham operation group (W-Sham group) and I/R group (W-I/R group).Seventy-two SPF male spontaneously hypertensive rats,aged 8 weeks,weighing 180-200 g,were randomly divided into 3 groups (n =24 each) using a random number table:sham operation group (S-Sham group),I/R group (S-I/R group),and curcumin group (S-Cur group).Global cerebral ischemia was induced by 4-vessel occlusion method (10 min of transient global ischemia followed by reperfusion).Curcumin 100 mg/kg was injected intraperitoneally at 30 min of reperfusion in S-Cur group,and corn oil 5 ml/kg was injected intraperitoneally at 30 min of reperfusion in the other groups.The ability of learning and memory was tested by step-down test at 7 days of reperfusion.Rats were sacrificed at 3 h and 1,3 and 7 days of reperfusion (T1-4) and the hippocampi were removed.The morphological changes of pyramidal cells in hippocampal CA1 area were observed by HE staining.The mean density of pyramidal cells in hippocampal CA1 area was quantified by Nissl staining.The contents of interleukin-lβ (IL-1β),tumor necrosis factor-alpha (TNF-α) and IL-10 in the hippocampus were determined by ELISA.Results Compared with W-Sham group,the ability of learning and memory was significantly decreased,the mean density of pyramidal cells in hippocampal CA1 area was decreased,the contents of IL-1β at T1-3,TNF-α at T1,and IL-10 at T2 were increased,and the contents of IL-10 were decreased at T1,3,4 in W-I/R group,and no significant changes were found in the parameters mentioned above in S-Sham group.Compared with W-I/R group,the ability of learning and memory was significantly decreased,the contents of IL-1β at T1,3 and IL-10 at T2 were decreased,the content of TNF-α was increased at T1,no significant change was found in the mean density of pyramidal cells in hippocampal CA1 area,and the damage to pyramidal cells in hippocampal CA1 area was severe in S-I/R group.Compared with S-I/R group,the ability of learning and memory was significantly increased,the mean density of pyramidal cells in hippocampal CA1 area was increased,the contents of IL-1β at T2,3 and TNF-α at T1-4 were increased,and the content of IL-10 was increased at T2,and the damage to pyramidal cells in hippocampal CA1 area was reduced in S-Cur group.Conclusion Inhibition of inflammatory responses in the hippocampus may be involved in the mechanism by which curcumin reduces global cerebral I/R injury in hypertensive rats.
9.Determination of atractylon in rat plasma by a GC-MS method and its application to a pharmacokinetic study
Han YAN ; Yuanyuan SUN ; Yuying MA ; Bin JI ; Xiaohong HOU ; Zhiguo YU ; Yunli ZHAO
Journal of Pharmaceutical Analysis 2015;5(5):327-331
A sensitive and selective method based on gas chromatography hyphenated to mass spectrometry (GC-MS) was developed and validated for the determination of atractylon in rat plasma. Plasma samples were processed by liquid-liquid extraction with ethyl acetate-n-hexane (1:1, v/v) using acetophenone as an internal standard (IS). Analytes were determined in selective ion monitoring (SIM) mode using target ions at m/z 108.1 for atractylon and m/z 105.1 for acetophenone. The calibration curve was linear over the concentration range of 10-1000 ng/mL with lower limit of quantification of 10 ng/mL. The intra- and inter-day precision variations were not more than 10.4% and 9.6%, respectively, whilst accuracy values ranged from -6.5% to 4.9%. Extraction recovery of the assay was satisfactory. This method was suc-cessfully applied to quantification and pharmacokinetic study of atractylon in rat plasma after in-tragastric administration of Atractylodis extract.