[Effect of twirling-reinforcing-reducing needling manipulations on contents of serum acetylcholine and arterial NOS and cGMP in stress-induced hypertension rats].

OBJECTIVE: To observe the effect of twirling-reinforcing or reducing needling manipulations on plasma acetylcholine (Ach) content and expression of nitric oxide synthetase (NOS) and cyclic guanosine monophosphate (cGMP) in thoracic artery tissue in stress-induced hypertension rats. METHODS: A total of 60 male rats were randomly divided into blank control, model, acupuncture (no-needle-manipulation) , twirling-reinforcing needling and twirling-reducing needling groups (n = 12 in each group). The stress hypertension model was established by giving the animals with noise and electric shock stimulation (paw), twice a day for 15 days. Acupuncture stimulation was applied to bilateral "Taichong" (LR 3) for 1 min, followed by retaining the needles for 20 min. The treatment was conducted once daily for 7 days. Systolic blood pressure of the rat's tail was detected with non-invasive method and plasma Ach, and NOS and cGMP contents in the thoracic artery tissue were measured using ELISA method. RESULTS: Compared with the control group, the systolic blood pressure was significantly higher in the model group after 15 days' stress stimulation (P < 0.01), while the contents of plasma Ach, arterial NOS and cGMP were markedly down-regulated (P < 0.01). Following 7 days' acupuncture interventions, the increased blood pressure was down-regulated in the no-needle-manipulation, twirling-reinforcing needling and twirling-reducing needling groups (P < 0.05, P < 0.01); and the decreased Ach and NOS in the 3 treatment groups, and cGMP levels in the twirling-reinforcing and twirling-reducing needling groups were remarkably up-regulated (P < 0.01, P < 0.05). No significant change of arterial cGMP content was found in the no-needle-manipulation group (P > 0.05). The effect of the twirling-reducing needling was superior to that of no-needle-manipulation and twirling-reinforcing needling in lowering blood pressure and raising plasma Ach content (P < 0.05, P < 0.01). CONCLUSION: The twirling-reducing needling of acupuncture has a significant anti-hypertensive effect in stress hypertension rats, which may be associated with its effects in raising blood Ach, and arterial NOS and cGMP levels.

[Effect of Yishen Daluo Decoction on the expression of PLP, Olig1, and Olig2 in mice with experimental autoimmune encephalomyelitis].

OBJECTIVE: To study the effect of Yishen Daluo Decoction (YDD) on the expression of protein lipoprotein (PLP), oligodendrocyte transcription factor 1 (Olig 1), and oligodendrocyte transcription factor 2 (Olig2) in mice with experimental autoimmune encephalomyelitis (EAE). METHODS: Totally 40 mice were randomly divided into 4 groups, i.e., the normal group, the model group, the Chinese medicine (CM) group, and the Western medicine (WM) group, 10 mice in each group. Each mouse in the model, CM, and WM groups was subcutaneously injected with 200 microL antigen emulsion (containing 150 micro g PLP139 -151 and 400 micro g H37RA) in two parts at the upper abdomen on the first day. 100 microLBordetella pertussis juice (containing 0. 6 x 10(6) Bordetella pertussis) was injected by caudal vein on the first and the third day. On the 7th day after modeling, each mouse in the normal group and the model group was intragastrically given normal saline (0. 1 mL/10 g). YDD (0. 2 g crude drug/10 g) was intragastrically given to mice in the CM group, and prednisone (0. 039 mg/10 g) was intragastrically given to mice in the WM group. All mice were intervened for 54 days. Changes of PLP, Olig1, and Olig2 in the brain tissue of EAE mice were detected by Western blot. Results The levels of PLP and Olig2 in the brain tissue of the model group were less than those of the normal group (P <0.05). Compared with the model group, the levels of PLP, Olig1, and Olig2 in the brain tissue increased in the CM group (P <0.05); the levels of PLP and Olig2 in the brain tissue increased in the WM group (P <0.05). Compared with the WM group, the level of Olig1 in the brain tissue increased in the CM group (P <0.05). CONCLUSION: YDD could enhance remyelination by elevating the levels of Olig1 and Olig2 in the brain tissue of EAE mice.

[Correlation between pulmonary function impairment and levels of alpha1-antitrypsin in serum and colon of ulcerative colitis patients: a clinical research].

OBJECTIVE: To explore the mechanism of pulmonary involvement in ulcerative colitis (UC) patients by observing the correlation between pulmonary functions and levels of alpha1-antitrypsin (A1AT) in serum and colon tissue in UC patients. METHODS: Totally 90 patients with confirmed UC were assigned to different groups according to the extent of disease, the disease activity, the staging of severity, and course of disease. The serum level of A1AT in UC patients with different extent of disease, the disease activity, the staging of severity, and course of disease were compared. And 30 healthy volunteers were recruited as the control group. The serum renal and hepatic functions, pulmonary functions, and serum levels of A1AT were detected in the UC group and the control group. The correlation between A1AT and each pulmonary function index in UC patients was analyzed. The A1AT content in the colon tissue was detected with immunohistochemical assay in 20 UC patients as well as in 10 healthy volunteers. RESULTS: Of the 90 UC patients, 54 patients were accompanied with pulmonary function abnormality (60.0%), and 24 with extraintestinal manifestations (26.7%). Compared with the control group, the serum level of A1AT was significantly lower in the UC group (P < 0.05). The serum level of A1AT was significantly higher in those with proctitis than in those with distal colonitis and pancolitis (P < 0.05). The serum level of A1AT was lower in patients with the course of disease 5 years and more than 5 years than in those with the course of disease less than 5 years (P < 0.05). Vital capacity (VC), forced vital capacity (FVC), forced expiratory volume in one second (FEV1.0), total lung capacity (TLC), function residual volume (FRV), and the ratio of diffusion capacity for carbon monoxide of lung (DLCO) were much lower in those with proctitis than in those with distal colonitis and pancolitis (P < 0.05). The ratio of FVC was negatively linear correlated with the course of disease (r = -0.23, P = 0.018). There was a positive correlation between the serum level of A1AT and peak expiratory flow (PEF) (r = 0.22, P = 0.03). The level of A1AT in the colon tissue was obviously lower in the UC patients than in those of the control group (P < 0.05). Mild and moderate UC patients had increased levels of A1AT in the colon tissue, when compared with severe UC patients (P < 0.05). The level of A1AT in the colon tissue was higher in those with proctitis than in those with distal colonitis and pancolitis (P < 0.05). CONCLUSIONS: The prevalence of pulmonary function impairment was higher than other extraintestinal manifestations in UC patients. The pulmonary function test was helpful to screen the pulmonary impairment of UC patients. The A1AT level in the serum and the colon tissue obviously decreased in UC patients, indicating the pulmonary function impairment of UC patients might manifest as decreased A1AT levels correlated chronic airway inflammation, remodeling of airway, and obstructive changes.

Effect of extracts from Radix Ginseng, Radix Notoginseng and Rhizoma Chuanxiong on delaying aging of vascular smooth muscle cells in aged rats.

OBJECTIVE: To observe the effect of extracts from Radix Ginseng, Radix Notoginseng and Rhizoma Chuanxiong (EXT) on delaying vascular smooth muscle cells (VSMCs) aging in aged rats. METHODS: VSMCs were obtained by the modified tissue explants technique and were shown to be positive for smooth muscle α-actin (SM-α-actin) by immunohistochemistry staining. VSMCs obtained from the young rats were served as the young control group; VSMCs obtained from the old rats were treated with no drug (the old group), with low dose extracts (20 mg/L, the EXT low-concentration group) and high dose extracts (40 mg/L, the EXT high concentration group), and with Probucal (10(-6) mol/L, the Probucal group) as a positive control. All groups were cultured for 24 h in the medium with 10% serum for 24 h followed by another 24 h in the serum-free medium. At the end of the 48-h culture, the following analyses were performed including determination of senescence-associated ß-galactosidase (SAß-Gal) activity, flow cytometry analysis of cell cycle, real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) analyses of p16, Cyclin D1, cyclin-dependent kinase 4 (CDK4) and retinoblastoma (Rb) mRNA expression, and Western blotting analyses of p16, cyclin D1, CDK4 and phosphoretinoblastoma (pRb) protein expressions. RESULTS: (1) In comparison to the younger rats, VSMCs from aged rats had significantly more SAß-Gal positive cells (P<0.01) and more cells in S phase (P<0.05). VSMCs from the all treated groups showed a significant decrease in both SAß-Gal positive cells (P<0.05) and S phase (P<0.05) compared to the old rats. (2) Compared with the young group, VSMCs in the old group had a significant decrease in p16 and Rb mRNA expression and a significant increase in Cyclin D1 and CDK4 mRNA expression. Compared with the old group, VSMCs in the treated groups had a significant increase in p16 and Rb mRNA expression and a significant decrease in Cyclin D1 and CDK4 mRNA expression (P<0.05). (3) Compared with the young group, VSMCs in the old group had a significant decrease in p16 protein expression and a significant increase in Cyclin D1, CDK4 and pRb protein expressions (P<0.05). Compared with the old group, VSMCs in the treated groups had a significant increase in p16 protein expression and a significant decrease in cyclinD1, CDK4 and pRb protein expressions (P<0.05). CONCLUSIONS: VSMCs obtained from old rats showed typical signs of cellular senescence and vascular aging. EXT had an effect on delaying senescence of VSMCs in vitro by altering the p16-cyclinD/CDK-Rb pathway.

[Effect of ginseng-sanqi extract on the Ras associated signal proteins].

OBJECTIVE: To explore the effect of ginseng-sanqi extract (GSE) on the vascular endothelium growth factor receptor-2 (VEGFR-2), Ras and mitogen-activated protein kinase (MAPK) in VEGFR-2-Ras-MAPK signal pathway of angiogenesis in cultured human umbilical vascular endothelial cells (HUVECs). METHODS: The protein expressions of VEGFR-2, Ras and MAPK in HUVEC and the effects of GSE (at different doses) and basic fibrin growth factor (bFGF) on them were detected by Western-Blot test. RESULTS: GSE can enhance the expression of angiogenesis signaling proteins (VEGFR-2, Ras, MAPK) to different extents, especially in the groups treated by bFGF or higher dose GSE, the levels showed significant differences to those in the blank group (P<0.05). CONCLUSION: GSE can promote the expressions of VEGFR-2, Ras and MAPK in the angiogenesis signaling pathway, which may be the foundation of Chinese medicine for tonifying qi and activating blood circulation in promoting endothelial proliferation and angiogenesis.

[Effects of huoxue injection on the adherence of human monocytes to endothelial cells and expression of vascular cell adhesion molecules].

OBJECTIVE: To observe the effect of Huoxue Injection (HXI, a Chinese herbal preparation consisted of red sage, chuanxiong, safflower and red peony root) on the expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), and the adherence of monocytes to endothelial cells in cultured human umbilical vein endothelial cells (HUVECs) injured by oxidized low-density lipoprotein (ox-LDL). METHODS: Model of injured cell was established by adding ox-LDL into the culture of HUVECs, and the model cells were intervened with HXI. The adhesive percentage of the model cells to monocytes was determined by protein quantification; mRNA and protein expressions of ICAM-1 and VCAM-1 were determined by RT-PCR and flow cytometry respectively. RESULTS: After HUVEC being treated with ox-LDL for 12 h and 24 h, its adhesion rate to monocytes increased, with the mRNA and protein expressions of ICAM-1 and VCAM-1 in HUVEC enhanced significantly, showing significant differences as compared with those in the normal control (P < 0.05 or P < 0.01). HXI could significantly reverse the above-mentioned changes dose-dependently, showing that these parameters in the HXI intervened cells significantly different to those in the untreated model cells respectively. CONCLUSION: HXI could inhibit the adherence of endothelial cells to monocytes by way of down-regulating the endothelial superficial adhesion molecules, so as to display its protection on endothelial cells, which should be helpful for reducing or suppressing the formation of atherosclerosis.

[Effects of tetramethylpyrazine on angiotensin II -induced proliferation and type I collagen synthesis of rat cardiac fibroblasts].

OBJECTIVE: To observe the effects of tetramethylpyrazine (TMP) on the proliferation and type I collagen synthesis of rat cardiac fibroblasts (CFBs) induced by angiotensin II (Ang II), and to explore the mechanism of TMP in treating myocardial fibrosis. METHODS: CFBs were isolated from neonatal rats, and the fourth-passage CFBs were used in the entire test and were stimulated by 0.1 micromol/L Ang II in vivo. The CFB proliferation was measured by methyl thiazolyl tetrazolium (MTT) assay. Type I collagen in the cell culture supernatant was measured by enzyme-linked immunosorbent assay. The expression of mRNA of type I collagen was semi-quantitatively measured by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: (1) In MTT assay, the optical density of CFBs cultured with 0.1 micromol/L Ang II was higher than that of the blank control cultured with 2% fetal bovine serum-Dulbecco's modified Eagle's medium (FBS-DMEM). The difference was statistically significant (P < 0.05). Both optical densities of CFBs cultured with 0.1 micromol/L Ang II plus 800 microg/mL TMP and 0.1 micromol/L Ang II plus 600 microg/mL TMP were lower than that of CFBs cultured with 0.1 micromol/L Ang II, but only the difference between 0.1 micromol/L AngII plus 800 microg/mL TMP group and 0.1 micromol/L Ang II group was significant (P < 0.05). (2) The content of type I collagen secreted by CFBs cultured with 0.1 micromol/L Ang II was higher than that with 2% FBS-DMEM (P < 0.01). The content of type I collagen secreted by CFBs cultured with 0.1 micromol/L Ang II plus 800 microg/mL TMP was lower than that with 0.1 micromol/L Ang II (P < 0.05). (3) The level of type I collagen mRNA in 0.1 micromol/L Ang II group was higher than that in blank control group, and lower than that in 0.1 micromol/L Ang II plus 800 microg/mL TMP group. Both the differences between 0.1 micromol/L Ang II group and the blank control group and between 0.1 micromol/L Ang II group and 0.1 micromol/L Ang II plus 800 microg/mL TMP group were statistically significant (P < 0.05). CONCLUSION: TMP can not only inhibit the proliferation of CFBs, but also decrease the secretion and the mRNA expression level of collagen I in cultured CFBs of rat which are increased by Ang II.

[Effects of Qingkailing effective components on nuclear factor-kappa B in an ischemia-reperfusion injury model of rat brain microvascular endothelial cells in vitro].

OBJECTIVE: To establish an ischemia-reperfusion injury model of rat cerebral microvascular endothelial cells (MVECs) in vitro, and to explore the relationship between nuclear factor-kappa B (NF-kappaB) and the protective effects of Qingkailing effective components (hyocholic acid, taurocholic acid, baicalin, jasminoidin, Pinctada martensii) on MVECs. METHODS: Brain MVECs of male rats were digested with trypsin and subcultured, then the content of MVECs was adjusted to 1x10 (5)/mL and the MVECs were divided into normal control group, untreated group, hyocholic acid group, taurocholic acid group, baicalin group, jasminoidin group, Pinctada martensii group and nimodipine group, with six holes in each group. Except for the normal control group, the MVECs in the other groups were exposed in oxygen and glucose deprivation (OGD) circumstance in vitro to simulate ischemia-reperfusion injury. Immunocytochemical staining and image analysis system were used to observe the expression of NF-kappaB protein. RESULTS: Under a light microscope, the nuclei of MVECs in the normal control group were blank. Staining intensity of NF-kappaB protein in the nucleus in the untreated group was much deeper than that in the endochylema, with NF-kappaB shifted to nucleus after activation; a small quantity of NF-kappaB protein were expressed in the border of nucleus next to endochylema in groups of Qingkailing effective components, and the NF-kappaB protein expression was weaker than that in the untreated group. With the image analysis, we found that transmittance of nucleus and endochylema in the untreated group was significantly lower than that in the normal control group (P<0.01). Transmittance of nucleus and endochylema in the treated groups was higher than that in the untreated group (P<0.05, P<0.01). CONCLUSION: Qingkailing effective components have significant effect in inhibiting NF-kappaB protein transferring from endochylema to nucleus in vitro.

[Effect of Huoxue injection on oxidized low-density lipoprotein induced activation of human umbilical vein endothelial cells].

OBJECTIVE: To observe effect of Huoxue injection on the expression of intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), the adherence of monocytes to endothelial cells, and the regulation role of nuclear factor kappa B (NF-kappaB) in cultured human umbilical vein endothelial cells (HUVEC) injury induced by the oxidized low-density lipoprotein (ox-LDL). METHOD: The ox-LDL (100 mg x L(-1)) was added to the cultured HUVEC to prepare the injury model of HUVEC. The adhesive percentage between HUVEC treated with ox-LDL and monocytes was determined by protein quantification. Expression of mRNA and protein of ICAM-1 and VCAM-1 were determined by RT-PCR and flow cytometry respectively. The percentage of positive cells and the ratio of nuclei and cytoplasm of NF-kappaB p65 staining in HUVEC the were examined by cell immunochemistry. RESULT: Treatment of HUVEC with ox-LDL for 12, 24 hours significantly increased adhesion of monocytes to HUVEC and enhanced the expressions of mRNA and protein of ICAM-1 and VCAM-1. The percentage of positive cells and the ratio of nuclei and cytoplasm of NF-kappaB p65 staining in HUVEC were significantly increased after treatment with ox-LDL for 24 hours. Huo Xue Injection could significantly inhibit the adhesion between monocyte and HUVEC, the expression of mRNA and protein of ICAM-1 and VCAM-1, and declined the percentage of positive cells and the ratio of nuclei and cytoplasm of NF-kappaB p65 staining in HUVEC. The effects were strengthened with increasing the deal of Huoxue injection. CONCLUSION: Huoxue injection has an inhibitory effect on the adherence of monocytes to HUVEC, probably by way of down-regulating the expression of mRNA and protein of ICAM-1 and VCAM-1 in HUVEC. The mechanism is probably associated with inhibiting the activation of NF-kappaB p65 of HUVEC. The effects of Huoxue injection can bring about the protective effect to endothelial cells injury induced by ox-LDL.

[Effect of three TCM methods for activating blood circulation on early stage apoptosis in rats with chronic atrophic gastritis complicated precancerous lesion].

OBJECTIVE: To comparatively study the effects of three TCM methods for activating blood circulation, i.e. in combined with resolving stasis (A), regulating qi (B) and supplementing qi (C), respectively, on early stage cell apoptosis in precancerous lesion of rats with chronic atrophic gastritis (CAG). METHODS: Rat model of CAG with precancerous lesion was duplicated by insertion of spring in pylorus and gastric perfusion of high-salt hot paste; and the impact of treatment on cell apoptosis was determined using Annexin V/PI double labeled flow cytometry. RESULTS: After being intervened for 12 weeks, the early stage cell apoptosis rate in the natural recovery group was significantly higher than that in the control group (P <0.01); while it lowered more significantly in the three groups receiving TCM therapeutic methods for activating blood circulation, showing significant difference compared with the natural recovery group (P <0.01). CONCLUSION: Three therapies of activating blood circulation all show inhibitory action on the early stage apoptosis of precancerous lesion in CAG rats, which is possibly one of their action mechanisms for improving or reversing the precancerous lesion.

[Effect of effective components of Huanglian Jiedu decocting on NF-kappaB in cultured rat cerebral microvascular endothelial cells injury induced by hypoxia and reoxygenation].

OBJECTIVE: To investigate the protective mechanism of geniposide, baicalin and berberine on hypoxia and reoxygenation injury in cultured rat cerebral microvascular endothelial cells. METHOD: To establish a model of hypoxia four hours and reoxygenation twelve hours injury in rat cerebral microvascular endothelial cells in vitro. The injured cells were treated with geniposide (0. 128, 0.064, 0.032 micromol mL(-1), baicalin (0.028, 0.014, 0.007 micromol mL(- 1)) and berberine (0.024, 0.012, 0.006 micromol mL(-1)). The expression of p65 subunit of NF-kappaB was detected by immunocytochemical assay and techniques of image quantitative analysis. The protein expression of NF-kappaB was calculated with the mean optical density and mean area. The nuclear translocation of NF-kappaB was calculated with the percentage of positive cells and ratios of light transmittance of cytoplasm and cell nucleus. RESULT: Compared with the normal group, both the protein expression and the nuclear translocation of NF-kappaB of model group were significant increased (P <0.01). Compared with the model group, the mean optical density of all treated groups was decreased ,but these was no significant difference between them. As compared with model group, the mean area of all treated groups was significant decreased (P < 0.01). The percentage of nuclear translocation of all treated groups is not only lower than that of the model group but higher than that of the normal group (P <0.01). Compared with the model group, the ratios of light transmittance of cytoplasm and cell nucleus of all treated groups was significantly elevated (P <0.01). CONCLUSION: The results suggesed that geniposide, baicalin and berberine could protect hypoxia/reoxygenation injuried rat cerebral microvascular endothelial cells injury. One of the mechanism may lie in inhibiting both the protein expression and the nuclear translocation of NF-kappaB.

Effects of Radix notoginseng extracts drug-containing serum on expressions of bcl-2, Bax and p21WAF1 proteins in MNNG transformed GES-1 cells.

OBJECTIVE: To investigate the effects of Radix notoginseng extracts drug-containing serum on the expressions of apoptosis-regulating proteins including Bax, bcl-2 and p21WAF1 in precancerous gastric cells. METHODS: The N-methyl-N'-nitro-N-nitroso-guanidine (MNNG) transformed eternalized human gastric mucosa epithelium GES-1 cell line (MC cell) was used in vitro as a model of gastric precancerous lesion. The medicated canine serum was prepared by feeding to the adult Beagle dog with Radix notoginseng extracts and obtaining the serum after 2-hour medication. MC cells were cultured with medicated canine serum (medicated serum group) or non-medicated canine serum (normal control group) for 72 hours. Expressions of Bax, bcl-2 and p21WAF1 proteins were detected by immunocytochemical assay and the average optical density of the cells was determined by an image analysis system. RESULTS: Compared with those of the normal control group, Bax and p21WAF1 expressions in medicated serum group were significantly enhanced (P<0.01), while the expression of bcl-2 was significantly reduced (P<0.01). CONCLUSION: Radix notoginseng extracts may inhibit the proliferation and promote the apoptosis of precancerous gastric cells through altering expressions of the bcl-2, Bax and p21WAF1 genes.

[Effect of effective components of huanglian jiedu decoction on hypoxia, reoxygenation injury and the expression of VCAM in cultured rat cerebral microvascular endothelial cells].

OBJECTIVE: To investigate the protective mechanism of geniposide, baicalin and berberine on hypoxia and reoxygenation injury in cultured rat cerebral microvascular endothelial cells. METHOD: A model of four hours hypoxia and twelve hours reoxygenation injury in rat cerebral microvascular endothelial cells in vitro was established. The injured cells were treated with geniposide (0.128, 0.064, 0.032 mmol x L(-1)), baicalin (0.028, 0.014, 0.007 mmol L(-1)) and berberine (0.024, 0.012, 0.006 mmol L(-1)), respectively. The immunocytochemical method and techniques of image quantitative analysis were used to detect the mean optical density and mean area in order to match the protein expression of VCAM-1. The method of RT-PCR was adopted to observe and match the mRNA expression of VCAM-1. RESULT: As compared with the normal group, the mean optical density, the mean area and the mRNA expression of VCAM-1 of model group were significant increased (P < 0.01, P < 0.01, P < 0.01). As compared with the model group, both the mean optical density and the mean area of all treated groups were decreased, and there was significant difference between them (P < 0.01, P < 0.01). As compared with normal group, the mean optical density of baicalin (0.007 mmol x L(-1)) and berberine (0.012, 0.006 mmol x L(-1)) were significant decreased (P < 0.05, P < 0.01, P < 0.01), but there was no significant difference between the other groups and the normal group. As compared with normal group, the mean area of baicalin (0.0014 mmol x L(-1)) was significant decreased (P < 0.05), but there was significant difference between the other groups and the normal group. The mRNA expression of all treated groups was not only lower than that of the model group but also higher than that of the normal group (P < 0.05, P < 0.05). CONCLUSION: The results suggest that geniposide, baicalin and berberine, which are effective compositions of huanglian jiedu decoting, can protect hypoxia-reoxygenation injuried rat cerebral microvascular endothelial cells. One of the protected mechanisms is that they can inhibit the expression of VCAM-1.

[Effects of panax notoginseng saponins on expression of intercellular adhesion molecule-1 in human umbilical vein endothelial cells injury induced by oxidized low-density lipoprotein].

OBJECTIVE: To study the protective effect of panax notoginseng saponins (PNS) on human umbilical vascular endothelial cells (HUVECs) injury induced by oxidized low-density lipoprotein (ox-LDL) for investigating the mechanism of PNS in treating arteriosclerosis obliterans (ASO). METHODS: Taking the cultured HUVECs as target cells, ox-LDL was used to establish a model of injured HUVEC and it was then intervened by PNS. The morphologic changes of HUVEC were observed under light microscope; activity of cells was determined by MTT method; the adhesive percentage between ox-LDL treated HUVEC and monocyte detennined by protein quantification and the protein expression of intercellular adhesion molecule-1 (ICAM-1) determined by flow cytometry. RESULTS: At the time points of HUVEC being treated with ox-LDL (100 mg/L) for 12 h and 24 h, significant injury of HUVEC was shown, its activity reduced, the adhesion rate with monocytes elevated, and the protein expression of ICAM-l in HUVEC increased significantly (P < 0.05, P < 0.01). PNS showed significant effect in reversing all the above changes, as compared with the control group (without PNS treaded), respective significant difference was shown in all the four indexes (P < 0.05, P < 0.01). CONCLUSION: PNS has a protective effect on endothelial cells injury induced by ox-LDL,which may be one of its mechanisms in treating ASO. The protective effect of PNS is probably by way of down-regulating the expression of ICAM-1 in endothelial cells and inhibiting the adherence of monocytes to endothelial cells.

[Comparative study on early application of the recipe for activating blood circulation and the recipe for supplementing qi for inhibiting left ventricular remodeling and apoptosis in rats with heart failure].

OBJECTIVE: To compare the characteristics of early application of the recipe for activating blood circulation and the recipe for supplementing qi for inhibiting left ventricular remodeling and apoptosis in rats with heart failure. METHOD: The left coronary artery occlusion was conducted to establish the rat model of heart failure after myocardial infarction. The model rats were randomly divided into 5 groups, including model group, activating blood circulation group (8 g x kg(-1)), supplementing qi group (8 g x kg(-1)), activating blood circulation plus supplementing qi group (16 g x kg(-1)), and captopril group (10.125 g x kg(-1)), and a sham operation group was set up as negative control group. The drugs were administrated on the second day after myocardial infarction with a therapeutic course of 4 weeks or 8 weeks. The heart function was detected by impedance method; Pathological staining and image analysis were used to determine the perimeter and the area of left ventricular cavity, and myocardial nuclei number and collagen content per unit area; Apoptosis percentage of the myocardial cell was detected by TUNEL and the content of Ang II in the cardiac muscle was measured by radioimmunoassay. RESULT: In comparison with the model group, the function of left ventricular contraction function improved, the area of left ventricular cavity diminished, and proliferation of collagen, content of Ang II and apoptosis percentage of the myocardial cell reduced in all of the treatment groups (P < 0.05 or P < 0.01). After treatment of 8 weeks, the activating blood circulation group was similar to the sham operation group in improvement of cardiac index, and decreases of the area of left ventricular cavity and the content of Ang II; Apoptosis percentage of the myocardial cell in the activating blood circulation group was significantly lower than that in the supplementing qi group. CONCLUSION: Both the recipes for activating blood circulation and for supplementing qi can inhibit left ventricular remodeling and myocardial apoptosis, and delay development of heart failure, with the best effect in the activating blood circulation group after treatment of 8 weeks.

[Protective action of effective components of Huanglian Jiedu decoction on hypoxia and reoxygenation injury in cultured rat cerebral microvascular endothelial cells].

OBJECTIVE: To investigate the protective effect of geniposide, baicalin and berberine for the rat cerebral microvascular endothelial cell. METHOD: The model of hypoxia and reoxygenation injury in rat cerebral microvascular endothelial cells in vitro was established. Both normal and model cells were treated with geniposide (1.024, 0.512, 0.256, 0.128, 0.064, 0.032, 0.016, 0.008 micromol x mL(-1)), baicalin (0.224, 0.112, 0.056, 0.028, 0.014, 0.007, 0.003 micromol x mL(-1)) and berberine (0.192, 0.096, 0.048, 0.024, 0.012, 0.006, 0.003 micromol x mL(-1)). Cell activity was measured by methyl thiazolyl tetrazolium (MTT) test. RESULT: After hypoxia/hypoglycemia cultures for 4 hour and reoxygenation for 12 hour, geniposide (0.128, 0.064, 0.032 micromol x mL(-1)), baicalin (0.028, 0.014, 0.007 micromol x mL(-1)) and berberine (0.024, 0.012, 0.006 micromol x microL(-1) could protect the injuried cerebral microvascular endothelial cells. CONCLUSION: Appropriate concentration of geniposide, baicalin and berberine, which are effective components of Huanglian Jiedu decoction, could protect the injuried cerebral microvascular endothelial cells.

[Cell proliferation inhibitive and apoptosis promoting effects of sanchi extract on GES-1 cell after being transformed by MNNG].

OBJECTIVE: To study the effect drug contained canine serum, prepared by gastric perfusion with Sanchi extract (SE), in inhibiting proliferation and promoting apoptosis of cultured precancerous gastric cells by cell culture. METHODS: The precancerous model cells (MC) used in the experiment were prepared through transforming eternalized human gastric mucosa epithelial cells GES-1 by N-methyl-N'-nitro-N-nitroso-guanidine (MNNG). After once gastric perfusion of SE extract to dogs, the canine serum gotten before and at different time points after medication was used for test. The inhibitory effect of the drug serum obtained at different time points on MC after acting for 72 hrs was detected by 3-(4,5)-dimethy thioazol-2-yl-2,5-diphenyl-tetrazoliumbromide (MTT) method to find the optimal time point for drug serum preparation, that were 2 hrs and 6 hrs after medication. Then the cell apoptosis promoting effect after acting for 72 hrs of the drug serum obtained at the optimal time points was determined by flow cytometry. RESULTS: The drug serum obtained at 2-hr and 6-hr after medication showed the highest inhibitive effect on MC cells, reaching 45.3% and 42.4% respectively, as compared with the effect of blank serum, the difference was significant (P<0.01). They could evidently promote the MC cell apoptosis, the apoptosis rate also showed significant difference to that of the blank serum (P < 0.05). Under their action, the proportion of MC cells in G0/G1 phase was obviously decreased (P < 0.05) while that in the G2/M phase significantly increased (P <0.05). However, the change of cells in S phase was not uniform. CONCLUSION: The drug contained canine serum gotten 2 hr and 6 hr after SE feeding shows the optimal MC proliferation inhibitive effect and significant apoptosis promoting effect. Besides, it could significantly decrease the proportion of MC cells in G0/G1 phase and significantly increase that in G2/M phase, this effect might be one of the mechanisms of ES in inhibiting MC cell proliferation and promoting its apoptosis.

[The protective effect of cholic acid on an in vitro injury model of ischemia-reperfusion in cerebral microvascular endothelial cells].

OBJECTIVE: To establish an in vitro injury model of ischemia-reperfusion in cerebral microvascular endothelial cells of rats and observe the protective effect of cholic acid. METHOD: Cultured rat microvascular endothelial cells were subjected to the oxygen-glucose deprivation (OGD) (Krebs solution) and recovery of oxygen-glucose, which simulated in vitro ischemia and reperfusion injury, and treated with cholic acid. The A value was measured with MIT chromatometry. RESULT: Cultured cells were impaired after OGD for 4 hours and recovery of oxygen-glucose for 12 hours, the A value of the cells treated with cholic acid was significantly higher than that of the cells without treatment (P < 0.01). CONCLUSION: Cholic acid could obviously protect rat cerebral microvascular endothelial cells from injury induced by an in vitro ischemia-reperfusion.

[Effect of danshensu and ligustrazine on related genes of myocardial hypertrophy induced by angiotensin II].

OBJECTIVE: To study the effect of Danshensu (DSS) and Ligustrazine (TMZ), the extracts of Chinese herbs for promoting blood circulation, on angiotensin II (Ang II) induced myocardial hypertrophy and its related genes, and to explore the mechanisms of inhibitory effect. METHODS: Adopting one-step method, the total RNA of myocardial cells was extracted by TRIzol reagent. Then the expression of ANP and beta-actin mRNA, as symbol of myocardial cells, were detected by RT-PCR. RESULTS: Molecular biological research showed that Ang II could significantly increase the expression of ANP mRNA in myocardial cells (P < 0.01), which could be significantly inhibited by Losartan (P < 0.01), both DSS and TMZ had the inhibitory effect (P < 0.05). Ang II could increase beta-actin mRNA expression in myocardial cells simultaneously, Losartan, DSS and TMZ could also significantly inhibit it (P < 0.05). CONCLUSION: The effective ingredients of Chinese herbs for promoting blood circulation, DSS and TMZ, have the effect of inhibiting the hyper-expression of ANP and beta-actin induced by Ang II, and preventing myocardial hypertrophy, therefore, it could be used to prevent and treat cardiomegaly.

[Apoptosis-promoting effect of Panax notoginseng extracts on MNNG-transformed GES-1 cells].

OBJECTIVE: To study the apoptosis-promoting effect of the serum from Panax notoginseng extracts-fed dog on precancerous gastric cells by means of flow cytometry. METHODS: In the experiment, we adopted eternalized human gastric mucosa epithelium GES-1 cells transformed by N-methyl-N'-nitro-N-nitroso-guanidine (MNNG) (MC cells) as the model of precancerous lesions for study in vitro. We took the serum of a dog before and at two different points of time (2 and 6 hours) after feeding the dog with Panax notoginseng extracts for experiment. The MC cells were cultured in mediums with different concentrations of the medicated serum at 2- or 6-hour point of time for 72 hours. By means of flow cytometry, we examined the apoptosis-promoting effects of the serums on the MC cells. RESULTS: The medicated serums at these 2 points of time had significant effects in promoting MC cell apoptosis. The proportions of apoptotic cells in culture mediums with medicated serums had a significant increase as compared with those in culture mediums with non-medicated serums (serum obtained before administration of extracts to the dog) under the same conditions (P<0.05). The number of MC cells in G(0)/G(1)phase was decreased (P<0.05) and that in G(2)/M phase increased (P<0.05), while no consistent changes were observed in S phase. CONCLUSION: The medicated serums obtained at the two different points of time have significant apoptosis-promoting effects on MC cells. They decrease the number of MC cells in G(0)/G(1) phase and increase the number of MC cells in G(2)/M phase. This is probably responsible for the effects of Panax notoginseng extracts in inhibiting the proliferation of MC cells and promoting its apoptosis.