[Effect of Modified Dihuang Yinzi Recipe and Huatan Tongluo Decoction on Neurological Function of MCAO Rats].

Objective To observe the effect of Modified Dihuang Yinzi Recipe (MDYR) and Hua- tan Tongluo Decoction (HTD) on neurological function of middle cerebral artery occlusion (MCAO) model rats. Methods Forty SD rats were randomly divided into 5 groups, i.e., the sham-operation group, the model group, the nimodipine (NMDP) group, the MDYR group, and the HTD group, 8 in each group. NMDP liquid was administered to rats in the NMDP group by gastrogavage at the daily dose of 12 mg/kg. MDYR liquid was administered to rats in the MDYR group by gastrogavage at the daily dose of 7. 9 mg/kg. HTD liquid was administered to rats in the HTD group by gastrogavage at the daily dose of 6. 5 mg/kg. Equal volume of distilled water as administered to rats in the sham-operation group and the model group by gastrogavage. All intervention lasted for 7 successive days. MCAO rat model was established. The Zealonga neurology score was measured. Neurological function was scored at 3 and 6 h, day 1 , 6, and 7, respectively. Levels of corticotropin releasing hormone ( CRH) in plasma, adrenocorticotropic hormone (ACTH) and cortisone (CORT) in serum were detected using radioimmunoassay. The expression of ma- trix metalloproteinase-9 ( MMP9 ) in brain tissue was detected using immunohistochemical staining. Results Compared with the sham-operation group, the Zea-longa neurological score increased; levels of CRH and ACTH decreased (P <0. 05, P <0. 01) , expression levels of CORT and MMP-9 (in brain tis- sue) increased in the model group (P <0. 01). Compared with the model group, the Zea-longa neurologi- cal score obviously decreased, levels of CRH and ACTH increased, expression levels of CORT and MMP-9 (in brain tissue) decreased (P <0. 05, P <0. 01) in the MDYR group and the HTD group. Com- pared with the NMDP group, serum CORT decreased in the MDYR group (P <0. 05) ; MMP-9 expression level decreased in the HTD group (P <0. 01). Conclusions MDYR and HTD could obviously improve neurologic function of MCAO rats. Its mechanism might be related to MDYR regulating disordered HPA ax- is and HTD inhibiting MMP-9 expression.

[Effect of chronic psychological stress on vascular endothelial dysfunction rats and intervention tongxinluo on it].

OBJECTIVE: To observe the effect of chronic psychological stress on vascular endothelial dysfunction rats and to explore the intervention and mechanism of Tongxinluo (TXL) on it. METHODS: Forty male Wistar rats were randomly divided into the normal control group (with no modeling), the endothelial dysfunction group (the HCY group), the psychological stress group (the model group), and TXL group, ten in each group. Rats in the latter three groups were fed with 3% high methionine diet to duplicate vascular endothelial dysfunction (VED) model. In addition, chronic psychological stress was applied in VED rats using repeated binding method. TXL at the dose of 1.2 g/kg body weight was given by gastrogavage. The plasma endothelin (ET) and angiotensin II (Ang II), serum cortisone (CORT) were detected by radioimmunoassay. The serum nitric oxide (NO) was detected by nitrate reductase. Ultrastructural changes of aortic endothelial cells were observed by transmission electron microscope. Serum levels of norepinephrine (NE) and epinephrine (E) were detected by enzyme linked immunosorbent assay (ELISA). RESULTS: Compared with the plasma ET level and the serum NO level in the HCY group (161.70 +/- 13.96 pg/mL and 26.82 +/- 13.03 micromol/L), the plasma ET level obviously increased (178.25 +/- 21.85 pg/mL) (P < 0.05) and the serum NO level decreased (24.91 +/- 9.95 micromol/L, P > 0.05), levels of CORT, NE, and E obviously increased in the model group (all P < 0.05). Ultrastructural changes of aortic endothelial cells were obviously injured. Compared with the model group, the plasma ET level (154.74 +/- 13.27 pg/mL), Ang II, CORT, NE, and E obviously decreased (P < 0.05, P < 0.01), the serum NO level obviously increased (34.44 +/- 18.35 micromol/L, P < 0.05). Ultrastructural changes of aortic endothelial cells were obviously improved. CONCLUSIONS: Chronic psychological stress could obviously aggravate endothelial injury in VED rats. TXL showed protection on the vascular endothelial structure and function.

Chinese medicine Tongxinluo modulates vascular endothelial function by inducing eNOS expression via the PI-3K/Akt/HIF-dependent signaling pathway.

AIM OF THE STUDY: To investigate the molecular mechanisms whereby the Chinese medicinal compound Tongxinluo improves vascular endothelial function through studying the induction of endothelial nitric oxide synthase (eNOS) and its upstream signaling pathway. MATERIALS AND METHODS: Hyperhomocysteinemia was induced in Wistar rats by a methionine-rich diet followed by Tongxinluo treatment. The aorta ring was isolated for measuring vascular dilation of aorta and eNOS expression. Human umbilical vein endothelial cells (HUVECs) were transfected with AP-1, NF-κB, HRE or eNOS reporter plasmid followed by Tongxinluo exposure. Expression of the reporter genes was measured by luciferase assay. The level of eNOS was studied by western blot and the nitric oxide content was measured using the nitrate reductase method. HUVECs were also transiently transfected with the dominant negative mutant of HIF-1, PI-3K or Akt to explore the role of HIF and PI-3K/Akt pathway in eNOS induction by Tongxinluo. RESULTS: Tongxinluo could significantly up-regulate the expression of eNOS in the aortic tissue and improve the endothelium-dependent vasodilation of the aorta ring. Additionally, Tongxinluo at various doses could significantly enhance the expression of HRE and eNOS reporter gene as well as up-regulate the protein level of eNOS. Meanwhile, Tongxinluo caused a dose-dependent increase in the NO content in the supernatant of HUVECs. Suppression of HIF-1 activation by DN-HIF or inhibition of PI-3K/Akt pathway by ΔP85 or DN-Akt both attenuated HRE reporter gene activation and eNOS induction by Tongxinluo. CONCLUSION: Tongxinluo, a compound Chinese traditional medicine, up-regulates the expression of eNOS via the PI-3K/Akt/HIF-dependent signaling pathway, thus improving the endothelium-dependent vasodilation.

[Effects of Tongxinluo on neuron ultrastructure and endothelial cell self-repairing ability in hypoxia preconditioning mice].

OBJECTIVE: To observe the ultrastructure changes of cerebral cortex neuron and endothelial cell in hypoxia preconditioning mice and the effects of Tongxinluo (TXL, Chinese traditional medilihe) on them. METHODS: Mice were randomly divided into 4 groups: control group, hypoxia group, hypoxia preconditioning (HP) group and Tongxinluo (TXL) group. The hypoxia preconditioning mice were exposed by repetitive hypoxia for 5 runs. The animal's tolerance time of each hypoxia run was recorded. The ultrastructure change of cerebral neuron and endothelial cell were studied by electron microscope. RESULTS: The hypoxic tolerance time in HP and TXL groups were significantly increased run by run. Compared with HP group, the tolerance time of TXL group were increased in every run. The ultrastructure of cerebral neuron and endothelial cell in hypoxia group changed obviously, mitochondrion and endoplasmic reticulum destroyed. However they were slighter in HP group than those in hypoxia group. The change in TXL group had no obvious differentce with control group and were slighter than those in HP group. CONCLUSION: Hypoxia preconditioning shows that organism has a strong self-repairing ability. Tongxinluo self-repairing; could increase self-repairing ability and adaptive ability of mice to hypoxia obviously.

[Effects of Tongxinluo on repeated hypoxic tolerance in mice].

OBJECTIVE: To observe the effects of Tongxinluo (TXL) on repeated hypoxic tolerance in mice and explore the underlying mechanisms. METHODS: Mice were randomly divided into groups of repeated hypoxia (control) and TXL according to body weights. The mice in each group were exposed to acute repeated hypoxia for 0 run (H0), 1 run (H1), 3 runs (H3) and 5 runs (H5). The animal's tolerance time of each hypoxic exposure was recorded. Western blot was used to measure the protein levels of hypoxia inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) in cortex tissue. RESULTS: The hypoxic tolerance time in control and TXL groups significantly increased run by run. Both HIF-1alpha and VEGF proteins in two groups increased gradually. Compared with control group, the tolerance time in H1 of TXL group [(18.0 +/- 2.4) minvs (15.6 +/- 2.0) min], H3 [(68.3 +/- 13.2) min vs (41.7 +/- 9.0) min) and H5 (85.9 +/- 7.0) min vs (51.4 +/- 14.4) min] increased (P < 0.05 or P < 0.01); the HIF-1alpha protein expression in H1 of TXL group (0.95 +/- 0.04 vs 0.79 +/- 0.02), H3 (1.01 +/- 0.03 vs 0.85 +/- 0.02), H5 (1.16 +/- 0.02 vs 0.92 +/- 0.03) increased (P < 0.05 or P < 0.01); the VEGF protein expression in H3 of TXL group (1.14 +/- 0.02 vs 0.89 +/- 0.03), H5 (1.34 +/- 0.05 vs 0.99 +/- 0.07) increased (P < 0.05 or P < 0.01). CONCLUSIONS: Under repeated hypoxia, an organism has a strong adaptive ability. The rises of HIF-1alpha and VEGF may be an adaptive mechanism. TXL can increase obviously the adaptive ability of mice to hypoxia.

[Extraction, combination and distribution regularity of syndrome elements in patients with variant angina pectoris].

OBJECTIVE: To explore the pathogenesis characteristics of variant angina pectoris (VAP) by extracting its syndrome elements and analyzing the combination and distribution regularity of the syndrome elements. METHODS: One hundred and seventy-five case files of VAP patients were collected. The extraction of syndrome elements and symptom contribution to syndrome was completed by the partition method of complex system based on entropy theory. Diagnostic threshold was established by receiver operator characteristic curve. According to the results diagnosed by diagnostic criteria for syndrome element with quantitation, the combination and distribution regularity of the syndrome elements in patients with VAP was analyzed. RESULTS: The basic syndrome elements in the patients with VAP were qi deficiency, qi stagnation, blood stasis, phlegm turbidity, phlegm-heat, stagnation-heat, yin deficiency and yang deficiency syndromes. It showed that the combination types of syndrome elements could be made up of one syndrome, two, three, four or more than four syndromes. Qi deficiency, yin deficiency, qi stagnation, blood stasis and phlegm turbidity syndromes had the higher frequency than other syndrome elements in the patients with VAP. CONCLUSION: The partition method of complex system based on entropy theory can be used in extracting the syndrome elements of the patients with VAP. It is found that VAP has complicated pathogenesis according to the combination and distribution regularity of syndrome elements. Qi deficiency, qi stagnation, blood stasis, phlegm turbidity and yin deficiency syndromes are the main syndrome elements.