Effect of Qufeng Tongluo Recipe on expression of desmin and CD2AP proteins in adriamycin-induced nephropathy rats: an experimental research / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of Qufeng Tongluo Recipe (QTR) on the expression of desmin and CD2-associated protein (CD2AP) in adriamycin-induced nephropathy rats.</p><p><b>METHODS</b>The adriamycin-induced nephropathy rat model was induced by a disposable intravenous injection of adriamycin. The model was successfully established after 3 weeks. Rats were then randomly divided into the blank control group (Group A, n =12), the model control group (Group B, n = 8), the small, medium, large dose QTR group (Group C, n = 8; Group D, n = 8; Group E, n = 8), and the positive control group (Group F, n = 8). From the fourth week normal saline was given to rats in Group A and Group B, QTR 1.0 g/mL, 2.1 g/mL, and 4.2 g/mL was respectively administered to those in Group C, D, and E. Prednisone 25 mg/kg was given to rats in Group F. All medication was performed by gastrogavage at 10 mL/kg, once daily, for 28 successive days. 24-h urinary protein excretion and sera biochemical indices were determined during medication. At the end of the experiment, ultrastructure was observed, mRNA expression of desmin, mRNA and protein of CD2AP were detected by Real-time PCR and Western blot.</p><p><b>RESULTS</b>(1) Compared with Group B, 24-h urinary protein excretion significantly decreased in Group C, D, E, and F (P < 0.05). (2) Compared with Group B, Alb in Group C, D, and E increased (P < 0.05) and TC significantly decreased (P < 0.05). TG significantly increased in Group F (P < 0.05). (3) Results of electron microscope showed, compared with Group B, the morphology of foot cells was improved to various degrees in Groups D, E, and F, especially the foot process structure and the number of foot processes were significantly improved, which was more obviously shown in Group D and Group E. (4) mRNA expression of desmin, mRNA and protein of CD2AP increased in adriamycin-induced nephropathy rats (P < 0.05). After intervention, when compared with Group B, mRNA expression of desmin and CD2AP were significantly lower in Group C, D, E, and F (P < 0.05). (5) Compared with Group A, expression of desmin and CD2AP significantly increased (P < 0.05). Compared with Group B, the expression of desmin protein were obviously lower in Group C, D, E, and F, and the protein expression of desmin obviously decreased in Group D, E, and F (P < 0.05). The protein expression of desmin and CD2AP gradually decreased in Group C, D, and E (P < 0.05). Compared with Group F, the expression of CD2AP protein obviously increased in Group C and D (P < 0.05); the expression of CD2AP protein obviously decreased in Group E (P < 0.05); the expression of desmin protein was higher in Group C, D, and E (P < 0.05).</p><p><b>CONCLUSION</b>QTR's therapeutic effect on adriamycin-induced nephropathy rats might be achieved through altered expression of desmin and CD2AP.</p>

Effects of controllable dynamic inhaled exposure of moxa smoke on LDL-r, ICAM-1 and morphology of heart tissue in rats / 中国针灸

<p><b>OBJECTIVE</b>To observe the change of lipid metabolism and vascular endothelium as well as morphology of heart tissue in rats who were long-time exposed to moxa smoke with different concentrations in order to provide reference for safety assessment of moxa smoke on cardiovascular system.</p><p><b>METHODS</b>One hundred and sixty-eighty Wistar rats were randomly divided into a control group, a low-concentration group, a median-concentration group and a high-concentration group, 42 rats in each one. The rats were exposed to moxa smoke with concentration of 0%, 10%, 40% and 70%, respectively, for 20 min per day. After continuous intervention for six months, enzyme-linked immunosorbent assay (ELISA) was applied to measure the level of low density lipoprotein-receptor (LDL-r) and intercellular adhesion molecule-1 (ICAM-1) in blood serum in each group; the slices of heart tissue were stained with hematoxylin-eosin staining method to observe morphology change of heart tissue.</p><p><b>RESULTS</b>(1) After the intervention of moxa smoke, the levels of LDL-r and ICAM-1 in the low-concentration group were not statistically different from those in the control group (both P > 0.05); the level of LDL-r in the median-concentration group was significantly increased, which was statistically different from that in the control group [(3.87 +/- 0.27) mg/mL vs (2.12 +/- 0.13) mg/mL, P < 0.01], however, the content of ICAM-1 was not obviously changed; although the level of LDL-r in the high-concentration group was presented with an escalating trend, it was not statistically different from that in the control group (P > 0.05) while the level of ICAM-1 was obviously increased (P < 0.01). (2) Under the light microscope, the abnormalities of cardiac muscle fibers and myocardial cell in each group were not been observed.</p><p><b>CONCLUSION</b>The long-time intervention of low-concentration moxa smoke has no significant effects on lipid metabolism and vascular endothelium of rats, indicating that clinical application of low-concentration moxa smoke is relatively safe. The long-time intervention of moderate-concentration moxa smoke could significantly increase the clearance rate of cholesterol, implying the beneficial regulation of moxa smoke on lipid metabolism. The high-concentration moxa smoke could induce certain damage to vascular endothelium but its mechanism is in need of further research. The pathologic change of heart tissue could not be induced by moxa smoke with any concentration.</p>

Effects and mechanisms of the functional parts of Dahuang Zhechong pill containing serum on platelet-derived growth factor-stimulated proliferation of vascular smooth muscle cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate and compare the effects and mechanisms of three functional parts of Dahuang Zhechong Pill (DHZCP), including drugs with the function of removing blood stasis and promoting blood circulation (FP-I), drugs with the function of expelling heat and moistening dryness (FP-II), and drugs with the function of nourishing yin and replenishing blood (FP-III) of DHZCP, on platelet-derived growth factor (PDGF)-stimulated vascular smooth muscle cells (VSMCs) proliferation with the method of serum pharmacology.</p><p><b>METHODS</b>VSMCs proliferation of rat was assayed by measuring the cell viability with the 3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyltetrazolium bromide (MTT) method. DNA synthesis in VSMCs was examined by detecting 5'-bromo-2'-deoxyuridine incorporation with the immunocytochemical method. Cycle of VSMCs was evaluated with flow cytometry. Expression of cyclin D1, p27, PKCα, and phosphorylated extracellular signal regulated kinase 1/2 (ERK1/2) was quantified by the Western blotting method.</p><p><b>RESULTS</b>The FP-I and FP-III containing serum was capable of inhibiting PDGF-stimulated proliferation and DNA synthesis of VSMCs, arrested VSMCs in G phase, downregulated cyclin D1, and upregulated p27 expression (P <0.01 or P <0.05). The FP-I and FP-III containing serum also inhibited the PDGF-induced phosphorylation of tyrosine of ERK1/2 and PKCα expression (P <0.01 or P <0.05).</p><p><b>CONCLUSIONS</b>FP-I and FP-III of DHZCP are able to inhibit VSMCs proliferation via interrupting PKCα-ERK1/2 signaling, modulating the expression of cell cycle proteins to result in arresting the cells in G phase. The inhibitory effect is mainly related to the function of removing blood stasis and promoting blood circulation, slightly to the function of nourishing yin and replenishing blood, but not to the function of expelling heat and moistening dryness.</p>

Dahuang zhechong pill containing serum inhibited platelet-derived growth factor-stimulated vascular smooth muscle cells proliferation by inducing G1 arrest partly via suppressing protein kinase C α-extracellular regulated kinase 1/2 signaling pathway / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate effects of dahuang zhechong pill ( DHZCP) on the cell cycle and the related signal pathways in vascular smooth muscle cells (VSMCs) stimulated by platelet-derived growth factor (PDGF) with the method of serum pharmacology.</p><p><b>METHODS</b>DNA synthesis in VSMCs was examined by detecting 5'-bromo-2'-deoxyuridine incorporation with the immunocytochemical method. The cycle of VSMCs was evaluated with flow cytometry. Expressions of cyclin D1, p27, protein kinase Cα (PKCα), and phosphorylated extracellular signal regulated kinase 1/2 (ERK1/2) were quantified by Western blot method.</p><p><b>RESULTS</b>DHZCP containing serum significantly inhibited DNA synthesis of PDGF-stimulated VSMCs, arrested the cells in G G(1) phase, modulated the protein expressions of cyclin D D(1) and p27, and suppressed the activation of PKCα and ERK1/2.</p><p><b>CONCLUSION</b>DHZCP containing serum inhibits VSMCs proliferation via modulating the expressions of cell cycle proteins to arrest the cell in G G(1) phase, which is attributed to, at least in part, suppressing PKCα-ERK1/2 signaling in VSMCs.</p>

Ellagic acid-induced hypercoagulable state in animals: a potentially useful animal hypercoagulable model for evaluation of anticoagulants.

OBJECTIVE: To establish and evaluate a hypercoagulable animal model for the assessment of anticoagulants. METHODS: Forty mice, thirty-two rats, and twenty-four rabbits were randomly and equally divided into control group (saline) and three ellagic acid (EA)-treated groups (low, middle, and high doses). In the mice, bleeding time (BT) was estimated with tail transaction, and clotting time (CT) with template method. Prothrombin time (PT) and the activated partial thromboplastin time (APTT) in rats and rabbits were measured by means of Quick's one-stage assay and modified APTT assay respectively. In addition, thrombin activity was estimated in rats with PT assay using a hemagglutination analyzer. The circulating platelet aggregates were detected in rabbits through platelet counting and presented as the circulating platelet aggregate ratio (CPAR). RESULTS: EA shortened BT and CT in mice, PT and APTT in rats, and increased thrombin activity and CPAR, all in a dose-dependent manner. EA also brought reduction of PT and APTT in rabbits in dose- and time-dependent manners. CONCLUSION: EA could induce hypercoagulable state through activating coagulation system and platelets in mice, rats, and rabbits.

Effect of Jinguo Weikang Capsule on proto-oncogene expression of gastric mucosa in rats with gastric precancerous lesions / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the effect of Jinguo Weikang Capsule [see text] on the gene expression of H-ras, epidermal growth factor receptor (EGFR), P53 and C-myc of the gastric mucosa in rats with gastric precancerous lesions, and to investigate the action mechanism of JWC on gastric precancerous lesions.</p><p><b>METHODS</b>A rat model with paratypical proliferation of the gastric epithelium mucosa was established by using 60Co irradiation. Rats were divided into the normal group, model group, high-, medium-, low-dose JWC treatment groups, and the vitacoenzyme control group, and were treated for 30 days. The expression of H-ras, EGFR, P53 and C-myc genes of the gastric mucosa was detected by using immunohistochemical methods.</p><p><b>RESULTS</b>The expression and over-expression rates of H-ras, EGFR, P53 and C-myc gene in the high-and medium-dose JWC treatment groups were significantly lower (P<0.05) as compared with those of the model group.</p><p><b>CONCLUSION</b>JWC can inhibit the expression of the H-ras, EGFR, P53 and C-myc genes expression of the gastric mucosa in rats, which may be one of mechanisms involved in suppressing or reversing gastric carcinogenesis.</p>

Study on anti-atherosclerotic mechanisms of divided functional recipes of dahuang zhechong pill in rabbits / 中国中药杂志

<p><b>OBJECTIVE</b>To study anti-atherosclerotic mechanisms of divided functional recipes of Dahuang Zhechong pill (DHZCP) in rabbits.</p><p><b>METHOD</b>The atherosclerotic rabbit model was established by high fat feeding combined with immune endothelial injury. Male New Zealand rabbits were divided into 9 groups: normal control group, model control group, Danshen positive control group, and 6 DHZCP-divided groups including divided functional recipes No. 1, 2, 3 with low and high doses for each divided recipe. After intragastric administration for 60 days, blood lipids and serum MDA and NO levels and SOD activity and plasma ET concentration, and contents of hydroxyproline and proteins in the vascular wall were determined.</p><p><b>RESULT</b>Compared with the model group, the level of blood lipids did not significantly change, serum MDA and ET levels, and the contents of hydroxyproline and proteins in the vascular wall significantly decreased (P < 0.05), and SOD activity and NO level increased in the divided functional recipes (all P < 0.05).</p><p><b>CONCLUSION</b>The divided functional recipes of DHZCP can inhibit development of atherosclerosis via a non-lowering lipid mechanisms, including anti-peroxidation of lipids, protection of endothelial function, and decrease of formation of extracellular matrix by reducing synthesis of collage and protein on the vascular wall. Among them, the divided functional recipe No. 1 exhibits the most obvious effect.</p>

Effect of the disassembled recipes of dahuang zhechong pill on proliferation and apoptosis of vascular smooth muscle cells in atherosclerotic rabbits / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effects of the disassembled recipes of Dahuang Zhechong Pill (DRDZP) on proliferation and apoptosis of vascular smooth muscle cell (VSMC) of thoracic aorta in atherosclerotic rabbits.</p><p><b>METHODS</b>The atherosclerotic rabbit model was induced by high-cholesterol diet and immune injury of endothelium. Expression of proliferating cell nuclear antigen (PCNA) in VSMC was detected by SP immunohistochemical technique, and VSMC apoptosis was observed with TUNEL technique.</p><p><b>RESULTS</b>Compared with the model control, PCNA expression in VSMC decreased, while apoptotic cells increased significantly in all the groups treated by DR-DZP (P < 0.01).</p><p><b>CONCLUSION</b>All the DR-DZP could inhibit VSMC proliferation and promote its apoptosis to modulate the balance between cell proliferation and apoptosis, so as to exert antiatherosclerotic action, among which the disassembled-recipe I is the main composition to contribute to antiatherosclerotic action of Dahuang Zhechong Pill.</p>

Anti-atherosclerotic effects of divided functional recipes of Dahang Zhechong pill in rabbits / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effects of the divided functional recipes of Dahuang Zhechong pill( DHZCP) on atherosclerosis in rabbits.</p><p><b>METHOD</b>The atherosclerotic model was established by the combination of hypercholesterol feeding and immune-injured endothelium in rabbits. Male New Zealand rabbits were randomly divided into nine groups: normal group, model group, Danshen group (0. 5 g x kg(-1) ), the low-dose(0. 5 g x kg(-1) ) and high-dose( 1.0 g - kg(-1) ) groups of the first divided recipe, the low-dose(0. 75 g x kg-' ) and high-dose(1. 5 g x kg(-1)) groups of the second divided recipe, the low-dose(0. 8 g x kg(-1) ) and high-dose( 1.6 g x kg(-1) ) groups of the third divided recipe. The effects of the divided functional recipes of DHZCP were observed in macropathology, histopathology and ultrastructure. Image analyzing system was used to determine atherosclerotic plaque area, intima thickness(IT) and intima-media thickness(IMT) in rabbit aorta.</p><p><b>RESULT</b>The divided functional recipes of DHZCP could significantly decreased the deposit of lipid and the atherosclerotic plaque area in aorta intima, relieve the histopathological changes of atherosclerosis, and inhibited the proliferation of vascular smooth muscle cells and collagen to reduce pachynsis of vascular intima. The divided functional recipes of DHZCP also reduced IT, IMT and IT/MT and reversed the contractive vascular remodeling.</p><p><b>CONCLUSION</b>The divided functional recipes of DHZCP produce the different anti-atherosclerotic action, among which the first divided functional recipe exhibits more effective action.</p>

Effect of ligustrazine on injury of vascular endothelial cells ECV-304 induced by hypoxia and lack of glucose / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effects of ligustrazine on nitric oxide (NO), malonaldehyde (MDA) production, release of intracellular lactate dehydrogenase (LDH) and membrane fluidity of the injured human umbilical vein vascular endothelial cell line (ECV-304) with hypoxia and lack of glucose.</p><p><b>METHOD</b>The experiments were performed in culture of ECV-304 injured with hypoxia and lack of glucose in vitro. The released LDH of ECV-304 was measured with automatic biochemistry analyse. NO content of ECV-304 was monitored with colorimetry. Lipid peroxidation of ECV-304 was monitored as MDA with a fluorometric assay. The membrane fluidity of ECV-304 was measured with the fluorescence polarization method.</p><p><b>RESULT</b>After culture ECV-304 in hypoxia and lack of glucose for 24 h, the LDH release, MDA production and the membrane fluidity increased significantly and NO level was decreased. Preincubation of ECV-304 with ligustrazine for 24 h reduced LDH release, MDA production, membrane fluidity increasing and increased the level of NO in ECV-304 due to hypoxia and lack of glucose.</p><p><b>CONCLUSION</b>Ligustrazine has protective effect on injury of ECV-304 induced by hypoxia and lack of glucose.</p>