Protective effect of Zige lyophilized powder on hypoxia/reoxygenation injury of human umbilical vein endothelial cells / 中国中药杂志

<p><b>OBJECTIVE</b>To study the protective effect of single prescription of traditional Chinese medicine Zige lyophilized powder on hypoxia/reoxygenation injury of human umbilical vein endothelial cells (HUVECs).</p><p><b>METHOD</b>HUVECs were cultured in vitro and Krebs liquid was adopted to establish the hypoxia/reoxygenation injury model. After intervention of Zige lyophilized powder, MTT colorimetric method was used to determine cell activity, the xanthine oxidase method is adopted to detect the activity of superoxide dismutases (SOD) in cells, the thibabituric acid developing method was adopted to determine the content of malondialdehyde (MDA), the nitrate reductase method was used to detect the content of nitric oxide (NO), the 2,4-dinitrophenylhydrazine developing method was adopted to the content of extracellular lactate dehydrogenase (LDH) , and the Western blot method was used to analyze the expression of cell apoptosis-related proteins Bcl-2, Bax and Caspase-3.</p><p><b>RESULT</b>Compared with the model group, 12.5 mg x L(-1) of Zige lyophilized powder can enhanced cell viability (P < 0.05). Besides, 49.0, 24.5, 12.5 mg x L(-1) of Zige lyophilized powder prevents the decline of SOD activity during H/R (P < 0.05). Furthermore, 24.5, 12.5 mg x L(-1) Zige lyophilized powder significantly inhibited the increase of MDA and NO content and improved the expression of Bcl-2 protein (P < 0.05, P < 0.01). And 49.0, 24.5, 12.5 mg x L(-1) of Zige lyophilized powder significantly decreased Bax level and up-regulated Bcl-2/Bax ratio. Caspase activation and apoptosis were monitored in the reoxygenated cells. 49.0, 24.5 mg x L(-1) Zige lyophilized powder can decreased Caspase-3 expression (P < 0.01).</p><p><b>CONCLUSION</b>Zige lyophilized powder can prevent H/R induced injury to HUVECs, which may be related to its effect in inhibiting cell peroxidation injury and enhancing cell antioxidant and antiapoptotic capacities</p>

Effect of catalpol and puerarin freeze-dried powder on coagulability, hemorheology and no in rats with Qi-deficiency and blood-stasis syndrome / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of catalpol and puerarin freeze-dried powder for injection (CPFPI), a new compound traditional Chinese medicine (TCM) preparation, on coagulability, hemorheology and NO in rats with qi-deficiency and blood-stasis syndrome.</p><p><b>METHOD</b>The model of rats with qi-deficiency and blood-stasis syndrome was established by hunger, fatigue, cold-dampness, panic and high fat diet. Coagulation time (CT) was observed by the glass method, and bleeding time (BT) was measured by tail-cutting method. The effects of CPFPI were also evaluated with prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time (TT). HCT was measured by the electric tesistance method, hemorheology indicators were observed by auto-hemorheological instrument. The level of NO in blood serum was measured by NO assay kit.</p><p><b>RESULT</b>CPFPI 65.40 mg x kg(-1) significantly prolonged CT, BT, PT, APTT and TT in rats. The viscosity of whole blood and plasma, hematocrit, erythrocyte aggregation and rigidity index, and reduced viscosity of whole blood in 65.40 mg x kg(-1) groups were lower than model group. CPFPI 65.40 mg x kg(-1) can raise the level of NO in blood serum. 32.70 mg x kg(-1) markedly prolonged CT, PT and APTT and decreased whole blood viscosity, erythrocyte aggregation index and whole blood reduction viscosity.</p><p><b>CONCLUSION</b>CPFPI has a significant effect in improving coagulability and hemorheology index and enhancing NO content in blood serum.</p>

Experimental studies on blood lipid regulating effects of shuanghua granules / 中国中药杂志

Hyperlipidemia plays a vital role in cardiovascular disease, and threatens our lives. The aim of this paper is to study the effects of Shuanghua granules on blood lipid in normal mice and different hyperlipidemia models. Acute and endogenous hyperlipidemia was induced in mice with yolk and Triton WR-1339 respectively. The model of hyperlipidemia in rats was set up by feeding high cholesterol diet. Then preventive effects of Shuanghua granules was observed compared with lovastatin and Zhibituo. We found that Shuanghua granules 5.6, 11.3, 22.5 g x kg (-1) could significantly reduce the serum TG level in normal mice (P < 0.01, P < 0.05). No significant difference was found in liver index, serum TG and HDL-C levels. When the mice were treated with either yolk or Triton WR-1339 in the presence of Shuanghua granules, the plasma lipoprotein levels (TC and LDL-C) were significantly reduced (P < 0.01, P < 0.05). Shuanghua granules could reduce the serum TC, TG, LDL-C, MDA, NEFA and liver TC, TG, LDL-C levels, simultaneously raise serum and liver HDL-C, serum SOD, LPL, HL, LA levels of hyperlipidemia rats (P < 0.01, P < 0.05). Shuanghua granules also significantly reduced whole blood viscosity, RV, etaP, IER and IEA (P < 0.01, P < 0.05), and lowered fatty degeneration of liver tissue. Compared with hyperlipidemia model, there was no significant increase in faeces lipoids concentrations. The results confirmed the mechanism of blood lipid regulating effects of Shuanghua granules is probably related with its antioxidation, regulating hemorheology and improving LPA, HL, LA enzymatic activity.

Progresses in research of hemolysis induced by puerarin injection / 中国中药杂志

Puerarin injection has been widely used for the treatment of cardiac/cerebral blood vascular diseases. The clinical statistical investigation had confirmed that puerarin injection can induce acute intravascular hemolysis, which effected the use of puerarin injection seriously. The review was focus on the research progress of the hemolysis of puerarin injection including the work in laboratory and clinic. The potential hemolysis mechanism and allergen of the puerarin injection were summarized and discussed.

Research of therapeutical effect and immunologic mechanism of Jiawei Foshou San on model rats of endometriosis / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the effect and mechanism of Jiawei Foshou San on the accretion ectopic endometrium of rats.</p><p><b>METHOD</b>Endometriosis model was established by surgical implant of endometrial tissue which belongs to its body in rats. Jiawei Foshou San was administrated to the model rats. Twenty-eight days later, the length of ectopic endometrium was measured by vernier caliper, the spleen was weighed by electronic balance, the content of tumor necrosis factor (TNF)-alpha in blood serum and peritoneal fluid was detected by ELISA test, the expression of interleukin (IL)-8 in ectopic endometrium was detected by immunohistochemical method, the expression of NF-kappaB p65 protein and inhibitory KBalpha (IkappaBalpha) protein in ectopic endometrium were analyzed by western blot.</p><p><b>RESULT</b>Jiawei Foshou San 0.045, 0.09, 0.18 g x kg(-1) group reduced the volume of ectopic endometrium. Jiawei Foshou San 0.18 g x kg(-1) group raised the spleen exponent of model rats. Jiawei Foshou San 0.09, 0.18 g x kg(-1) group decreased the content of TNF-alpha in blood serum and peritoneal fluid, and the content of IL-8 in ectopic endometrium was also decreased. Jiawei Foshou San can decrease the expression of NF-KB p65 and increase the expression of IkappaBalpha in ectopic endometrium.</p><p><b>CONCLUSION</b>Jiawei Foshou San can inhibit the accretion of endometriosis implants of rats, and its mechanism might be associated with improving the environment of body.</p>

Inhibitory effect on estrogen production and its influence on invasive ability of human endometrial cells of endometriosis by medicated serum of SLW / 中国中药杂志

<p><b>OBJECTIVE</b>To study the inhibitory effect of medicated serum of SLW on estrogen production and to approach on the key mechanism of SLW in inhibiting the invasion ability of endometrial cells of endometriosis.</p><p><b>METHOD</b>First, the model of eutopic primary cultured endometrial cells of endometriosis and hysteromyoma in vitro was successfully established. Taking that of endometrial cells of hysteromyoma as control, the secretion level of E2 of endometrial cells in the culture media supernatant at different time point with the treatment of high, middle and low dose of SLW serum was detected by electrochemiluminescence immunoassay, the activities of matrix metalloproteinase-2, 9 (MMP-2, 9) were detected by gelatinase zymography assay, and the expression of tissue inhibitor of metalloproteinase-1, 2 (TIMP-1, 2) protein was observed by immunofluorescence. After the optimal time for SLW to inhibit invasion ability of endometrial cells was identified based on time-effect relationship, another endometrial cells were divided into six groups: hysteromyoma endometrium group, eutopic endometrium of endometriosis group, eutopic endometrium of endometriosis + middle dose of SLW serum group, eutopic endometrium of endometriosis + middle dose of SLW serum + E2 group, eutopic endometrium of endometriosis + anastrozole serum group , and eutopic endometrium of endometriosis + E2 group. The activities of MMP-2, 9 and the expression of TIMP-1, 2 protein were detected according to the optimal time point.</p><p><b>RESULT</b>The secretion level of E2 of eutopic endometrium in endometriosis could be decreased by SLW, which showed the dependence of time and concentration. The result of gelatinase zymography assay and immunofluorescence respectively showed that along with the time the activities of MMP-2, 9 of eutopic endometrial cells of endometriosis were significantly higher than those of hysteromyoma at the same time point (P < 0.01). After 48 hours, with the treatment of middle dose of SLW serum, the activities of MMP-2, 9 of eutopic endometrial cells of endometriosis could be decreased (P < 0.01) while the expression of TIMP-1, 2 protein could be increased obviously (P < 0.01). The malignant invasion ability improved by SLW of eutopic endometrial cells of endometriosis was partly recruited by add-back E2 treatment. There was no significant difference in the activity of MMP-2 and the expression of TIMP-1, 2 protein between eutopic endometrium of endometriosis + middle dose of SLW serum + E2group and untreated group. The behavior of invasion of endometrial cells of endometriosis could be deteriorated treated by E2 as contrast to anastrozole, a specific aromatase inhibitor.</p><p><b>CONCLUSION</b>SLW could decrease the secretion of E2 so as to inhibit the invasion of the eutopic endometrial cells of endometriosis.</p>

Effects of Erbie San on Walker-256 liver cancer and adjustment to unbalance of VEGF/endostatin in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the anticancer effects of Erbie San on the rats bearing Walker-256 liver cancer and the potential mechanism of its angiogenesis effects.</p><p><b>METHOD</b>Wistar rats bearing Walker-256 liver cancer were used in this study. The experimental groups were treated with Erbie San 1.25, 2.5, 5 g x kg(-1) x d(-1), and 5-Fluorouracil injection 75 mg x kg(-1), respectively. The tumor's weight, the expression of VEGF, Endostatin and the ratio of VEGF/endostatin in serum of each groups were observed.</p><p><b>RESULT</b>Compared to the model group, Erbie San 2.5, 5 g x kg(-1) x d (-1) and 5-Fluorouracil injection groups can reduce the tumor's weight significantly (P<0.05 or P<0.01). The expression of VEGF was reduced, while endostatin was increased, and the ratio of VEGF/endostatin was reduced (P<0.05 or P<0.01).</p><p><b>CONCLUSION</b>Erbie San can effectively inhibit the growth of Walker-256 liver cancer of rats and can inhibit the expression of VEGF but increase the expression of endostatin, which suggest that Erbie San has the inhibition of angiogenesis which is responsible for its anticancer effects.</p>

Inhibitory effect of Sanlengwan on expression of adhesion molecules ICAM-1, CD44 in rats of endometriosis / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the effect of Sanlengwan (SLW) on adhesion molecules expression and the accretion in ectopic endometrium of rats.</p><p><b>METHOD</b>Endometriosis was established by surgical implant of endometrial tissue which belong to its body. Forty EMS model rats were randomly divided into five groups: model control group, three dose of SLW groups and anastrozole group. Meanwhile, eight normal rats were used as the normal group. All the rats were treated for 4 weeks respectively, and the volume of grafts were measured by vernier caliper, morphological changes were measured by HE stain, and the adhesion molecules, ICAM-1 and CD44 protein, were also measured by immunohistochemical test before and after treatment of SLW.</p><p><b>RESULT</b>SLW markedly reduced the volume of grafts, improve the morphological characters and decreased the expression of ICAM-1 and CD44 in ectopic endometrial tissue.</p><p><b>CONCLUSION</b>SLW can inhibit the accretion of ectopic endometrium tissue of rats, and its mechanism might be associated with inhibiting the expression of ICAM-1 and CD44 protein.</p>

Effect of rat serum containing different concentrations of tramethylpyrazine on proliferation of vascular endothelial cells in vitro / 中国组织工程研究

BACKGROUND: Studies have shown that 160 mg/L tramethylpyrazine (TMP) can inhibit the proliferation of endothelial cells cultured in vitro,but whether TMP also inhibits vascular endothelial growth factor (VEGF)induced proliferation of vascular endothelial cells remains unkown. OBJECTIVE: To study the effect of rat serum containing TMP on vascular endothelial cell proliferation and its proliferation induced by VEGF. DESIGN: Randomized controlled experiment. SETTING: College of Traditional Chinese Medicine, Chongqing University of Medical Sciences. MATERIALS: The experiment was carried out in the Institute of Pediatrics, Chongqing University of Medical Sciences from March, 2002 to March, 2003 using human umbilical vein endothelial cell (HUVEC) line ECV304 and 30 female Wistar rats. METHODS: Thirty rats were randomly divided into 3 equal groups to receive intraperitoneal TMP injection at 143.0 mg/kg and 71.5 mg/kg and 0.8 mL normal saline (blank control group). All the injections were performed once a day for consecutive 7 days, then blood was collected from the intraperitoneal artery and diluted to 20％, 10％, and 5％ in triplicate.Effect of TMP-containing serum on the proliferation of ECV304 cells was observed by means of in vitro culture and 3H-TdR incorporation as well as methyl thiazolyl tetrazolium (MTT) methods. MAIN OUTCOME MEASURES: ① Effect of TMP-containing serum on proliferation of ECV304 cells in in vitro culture and VEGF-induced proliferation of the cells.RESULTS:All the 30 rats survived the experiment without losses. The absorbance (A) and scintillation counting (minute-1) of the cells were significantly lower in 143.0 mg/kg TMP group than those in the control group treated with rat serum at the dilution of 5％ (0.720±0.024 vs 0.816±0.068,3340.45±567.7 vs 5120.84±301.49), 10％ (0.630±0.017 vs 0.798±0.015,2430.06±265.98 vs 5225.83±100.10), and 20％ (0.765±0.027 vs 0.823±0.031,3570.45±130.52 vs 5256.82±183.18), and those in 71.5 mg/kg TMP group were also significantly lower than those in the control group after treatment with the serum of 10％ (0.775±0.023, 4571.14±275.39) and 20％(0.749±0.012, 3287.25±144.82). In the experiment evaluating the effect of TMP serum on VEGF-induced proliferation of ECV304 cells, the A value and scintillation counting in the 143.0 mg/kg TMP group were significantly lower than those in the control group after the cells were treated with the serum of 5％ (0.726 ±0.004 vs 0.964 ±0.004, 5760.46 ±49.64 vs 9821.82±128.05), 10％ (0.712±0.004 vs 0.933±0.014, 5024.48±100.57 vs 9052.76±65.19), and 20％ (0.717±0.003 vs 0.924±0.004, 5405.45±140.90vs 9197.07±169.92], and those of the cells treated with 71.5 mg/kg TMP serum of 10％ and 20％ were also significantly lower than those in the control group (0.703±0.005 and 7526.47±169.21 for 10％ serum, and 0.693±0.006 and 5720.09±279.03 for 20％ serum). CONCLUSION: The sera at the concentrations of 5％,10％, and 20％from rats treated with 143 mg/kg TMP and at higher concentrations from 71.5 mg/kg TMP-treated rats can inhibit the proliferation of ECV 304 in in vitro culture as well as VEGF-induced proliferation of the cells.