Huoxue Anxin Recipe () promotes myocardium angiogenesis of acute myocardial infarction rats by up-regulating miR-210 and vascular endothelial growth factor / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the microRNAs (miRNAs) expression profile of acute myocardial infarction (AMI) rats and the regulating effects of Huoxue Anxin Recipe (, HAR) on angiogenesis-related miRNAs and genes.</p><p><b>METHODS</b>Forty-five Wistar rats were randomly assigned to 3 groups according to a random number table: sham, AMI, and AMI+HAR groups (15 in each group). AMI rats were established by ligation of the left descending coronary artery. HAR was intragastrically administered to rats of the AMI+HAR group for successive 21 days since modeling, meanwhile the same volume of 0.9% normal saline was administered to rats of the sham and AMI groups. Doppler echocardiography was used for noninvasive cardiac function test. Hematoxylin and eosin staining was used to observe the histopathological change. miRNAs expression profile was detected by quantitative realtime polymerase chain reaction (qRT-PCR). The mRNA and protein expressions of vascular endothelial growth factor (VEGF), and a target gene of miR-210 was further detected by qRT-PCR and Western blot, respectively. The microvessels density of myocardium was evaluated by CD31 immunostaining.</p><p><b>RESULTS</b>Compared with the sham group, ejection fraction (EF) and fractional shortening (FS) values were decreased significantly in the AMI group (P<0.01), while the infarction area and the interstitial collagen deposition were increased obviously. As for the AMI+HAR group, EF and FS values were increased significantly (P<0.05 vs. AMI group), and the infarction area was reduced and the interstitial collagen deposition were alleviated significantly. Total of 23 miRNAs in the AMI group expressed differently by at least 1.5 folds compared with those in the sham group; 5 miRNAs in the AMI+HAR group expressed differently by at least 1.5 folds compared with those in the AMI group. Among them, miR-210 was low in the AMI group and high in the AMI+HAR group. The relative mRNA and protein expressions of VEGF were decreased significantly in the AMI group (P<0.05 vs. sham group), and increased significantly in the AMI+HAR group (P<0.01 vs. AMI group). CD31 expression area and optical intensity were decreased significantly in the AMI group (P<0.05 vs. sham group), and increased significantly in the AMI+HAR group (P<0.01 vs. AMI group).</p><p><b>CONCLUSIONS</b>HAR could reduce the infarction area, alleviate the interstitial fibrosis and improve the cardiac function of AMI rats. Those effects could be related to promoting myocardium angiogenesis of HAR by up-regulating miR-210 and VEGF.</p>

Huoxue anxin recipe alleviated peroxidation damage of acute myocardial infarction rats by regulating iNOS/eNOS imbalance: an experimental research / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the protective mechanism of Huoxue Anxin Recipe (HAR) on peroxidation damage of acute myocardial infarction (AMI) rats.</p><p><b>METHODS</b>The AMI rat model was established by occluding the left anterior descending coronary artery. Compound Danshen Dripping Pill (CDDP) was used as the positive control. CDDP and HAR were administered to rats for 7 successive days since modeling. The heart function was detected using color Doppler echocardiography. Activities of induced nitric oxide synthase (iNOS), endothelial nitric oxide synthase (eNOS), total superoxide dismutase (tSOD) activity, and contents of malondialdehyde (MDA) were detected by ultraviolet spectrophotometer method.</p><p><b>RESULTS</b>Compared with the sham-operation group, ejection fraction (EF) and fraction shortening (FS) rate significantly decreased in the model group (P < 0.01). Compared with the model group, EF and FS rate significantly increased in the HAR group, showing statistical difference (P < 0.05). There was no statistical difference in activities of serum iNOS, eNOS, or tSOD among all groups (P > 0.05). Compared with the sham-operation group, iNOS activities and MDA contents significantly increased in the myocardial tissue of the model group (P < 0.01), activities of eNOS and tSOD significant decreased (P < 0.01). Compared with the model group, iNOS activities in the myocardial tissue, MDA contents both in serum and the myocardial tissue significantly decreased (P < 0.05), activities of eNOS and tSOD significantly increased in the HAR group (P < 0.05). There was no statistical difference in each index between the CDDP group and the HAR group (P > 0.05).</p><p><b>CONCLUSIONS</b>HAR could significantly improve cardiac functions of AMI rats. Its roles might be associated with regulating imbalanced iNOS/eNOS expressions and alleviating peroxidation damage of the myocardial tissue.</p>

The protective mechanism study of fengshiqing recipe against bone destruction in CIA rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the protective mechanism of Fengshiqing Recipe (FR) against bone destruction in collagen-induced arthritis (CIA) rats.</p><p><b>METHODS</b>Rats were divided into four groups in the experiment,i.e., the blank control group, the model group, the MTX group (MTX, 1 mg/1 000 g), and the FR group (24 g crude FR/kg). The CIA model was prepared except the blank control group. Medication was started in the MTX group and the FR group from the 14th day after modeling to the 56th day. The toe volume was measured on every Tuesday and Friday. Expression levels of serum IL-17, RANKL, MIP-1alpha were detected after 3-and 6-week intervention. The bone scintigraphy with nuclide (SPECT), bone mineral density (BMD), and the pathological section were observed to assess the intervention of drugs of heat clearing blood activating actions in the bone destruction of CIA rats.</p><p><b>RESULTS</b>From the 10th day of modeling, the volume of both toes started to swell and reached the peak at about 21 days. It was obviously shrunk at about 30 days. Of them, the swelling degree was milder in the MTX group and the FR group than in the model group. Compared with the model group at the same phase, the levels of IL-17 and RANKL decreased in the MTX group after 3 weeks of intervention (P < 0.01, P < 0.05). The IL-17 level decreased in the FR group after three weeks of intervention (P < 0.05). The RANKL level decreased in the MTX group and the FR group after 6 weeks of intervention (P < 0.01, P < 0.05). Compared with the model group and the MTX group, the overall BMD and ankle BMD increased in the FR group after 6 weeks of intervention (P < 0.01, P < 0.05). The ankle ROI/mandible and the toe ROI/mandible were elevated in the FR group after 3 weeks of intervention (P < 0.05). Pathological results suggested that the joint lacunae was significantly widened, the hyperplasia of the synovial tissue was so severe, and the bone tissue was destroyed in the model group. Compared with the model group, the aforesaid conditions were significantly improved in the MTX group and the FR group. The cartilage structure was complete.</p><p><b>CONCLUSION</b>QR could inhibit decreased BMD, prevent bone destruction, which might be achieved by down-regulating expression levels of IL-17, RANKL, and MIP-1alpha through the osteo immunological Th/RANKL system,inhibiting maturation and differentiation of osteoclasts, thereby, inhibiting bone destruction.</p>

Salvianolic acid B inhibits the TLR4-NFκB-TNFα pathway and attenuates neonatal rat cardiomyocyte injury induced by lipopolysaccharide / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the role of the TLR4-NFκB-TNFα inflammation pathway on: lipopolysaccharide (LPS)-induced neonatal rat cardiomyocyte injury and the possible protective effects of salvianolic acid B (Sal B).</p><p><b>METHODS</b>Wistar rat (1-2 days old) cardiomyocytes were isolated and cultured. Sal B 10(-5)mol/L, 10(-6)mol/L and 10(-7)mol/L were pre-treated for 6 h in the culture medium. LPS (1 μg/mL) was added to mol/the culture medium and kept for 6 h to induce inflammation injury. The concentration of lactate dehydrogenase (LDH) in the supernatant was detected by spectrophotometry. The concentrations of tumor necrosis factor α (TNFα) and heat shock protein 70 (HSP70) in the supernatant were detected by enzyme linked immunosorbent assay. The protein expressions of toll, such as receptor 4 (TLR4) and nuclear factor kappa B (NFκB) were detected by immunohistochemistry. The mRNA expressions of TLR4 and NFκB were detected by real-realtime reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>(1) The concentrations of LDH and: TNFα in the LPS control group were significantly higher than those in the control group (561.41±67.39 U/L and 77.94±15.08 pg/mL, versus 292.13±26.02 U/L and 25.39±16.53 pg/mL, respectively, P<0.01, P<0.05). Compared with the LPS control group, the concentrations of LDH and TNFα were significantly decreased in the Sal B 10(-5)mol/L pre-treated group (451.76±83.96 U/L and 34.00±10.38 pg/mL, respectively, P<0.05). (2) The TLR4 and NFκB protein expression area in the LPS control group were significantly higher than those in the control group (1712.41±410.12 μm(2) and 2378.15±175.29 μm(2), versus 418.62±24.42 μm(2) and 1721.74±202.87 μm(2), respectively, P<0.01). The TLR4 and NFκB protein expression internal optical density (IOD) values in the LPS control group were also significantly higher than those in the control group (3.06±0.33 and 7.20±1.04, versus 0.91±0.21 and 4.24±0.48, respectively, P<0.05 and P<0.01). Compared with the LPS control group, the TLR4 and NFκB protein expression areas were significantly decreased in the Sal B 10(-5)mol/L pre-treated group (1251.54±133.82 μm(2) and 1996.37±256.67 μm(2), respectively, P<0.05), the TLR4 and NFκB protein expression IOD values were also significantly decreased in the Sal B 10(-5)mol/L pre- mol/pretreated group (1.92±0.28 and 5.17±0.77, respectively, treated P<0.05). (3) The TLR4 and NFκB mRNA expressions (2(-ΔΔ)CT value) in the LPS control group were significantly higher than those in the control group (3.16±0.38 and 5.03±0.43 versus 1.04±0.19 and 1.08±0.21, respectively, P<0.01). Compared with the LPS control group, the TLR4 and NFκB mRNA expressions (2(-ΔΔ) -CT value) were significantly decreased in the Sal B 10(-5)mol/L pre- mol/pretreated group (1.34±0.22 and 1.74±0.26, respectively, treated P<0.05). The concentration of HSP70 did not show any <statistical differences in all groups (P>0.05).</p><p><b>CONCLUSIONS</b>The TLR4-NFκB-TNFα pathway was quickly activated: and was independent of HSP70 in the early phase of neonatal cardiomyocyte injury induced by LPS. The protective effects of Sal B may be through inhibiting the TLR4-NFκB-TNFα pathway and are dose-dependent.</p>

Variation of TFAR-19 protein expression in the thymus of emotional stress mice and the regulatory effect of Modified Xiaoyao Pill / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the variation of TFAR-19 protein expression and Annexin-V apoptosis protein proportion in the mice thymus cell apoptosis procedure induced by water platform environment or electric emotional stress and the regulatory effect of Modified Xiaoyao Pill (MXP).</p><p><b>METHODS</b>The mouse emotional stress model was established by water platform equipment or electrical stimulation. The serum glucocorticoid was detected by radioimmunoassay, the TFAR-19 protein was detected by flow cytometry analysis, and Annexin-V apoptosis protein proportion was calculated by immunofluorescence technique.</p><p><b>RESULTS</b>In the groups of mouse stressed by water platform environment, the level of serum glucocorticoid, the TFAR-19 protein expression and the Annexin-V apoptosis protein proportion increased in the thymus cell along with the stress time prolonging (P<0.05 or P <0.01). The serum glucocorticoid level in mice treated with MXP was lower than that in the untreated group (P <0.05). In the groups of emotional stressed mouse established by electrical stimulation, the above-mentioned variations also revealed. All these variations could be alleviated with MXP (P<0.05).</p><p><b>CONCLUSION</b>The water platform environment stress is a chronic continuous stress and electrical stimulation is an acute smooth stress, both of them could damage thymus function through neuro-endocrineo-immune network, but different in duration for causing severe injury. Chinese medicine MXP can alleviate the damage of thymus induced by either of them to certain degree.</p>