Mechanism of Wenxin Granule Inhibits Myocardial Cell Apoptosis After Myocardial Infarctionby Regulating Endoplasmic Reticulum Stress in Rats / 中国实验方剂学杂志

Objective:To explore the intervention effect of Wenxin granule on endoplasmic reticulum stress apoptosis pathway and its related mechanism in rats with myocardial infarction. Method:Ligating the left anterior descending coronary artery to establish the rat model， the rats were randomly divided into 5 groups， namely sham group， model group， betaloc group and low， high-dose Wenxin granule groups，10 in each group. The sham group and the model group were given 10 mL∙kg^-1∙d^-1 deionized water， the low， high-dose modified Wenxin groups were given 1.35， 2.7 g∙kg^-1∙d^-1 aqueous solution respectively， and the betaloc group was given 2.25 mg∙kg^-1∙d^-1 aqueous solution. After 14 days，the catheter method was used to detect the cardiac hemodynamics and hematoxylin-eosin staining （HE） was used to observe the pathological morphological changes. The levels of endoplasmic reticulum stress protein glucose regulatory protein 78（GRP78）， protein kinase R like endoplasmic reticulum kinase（PERK）， phosphorylated activated PERK（p-PERK）， activated transcription factor 6 （ATF6）， nuclear transcription factor X cassette binding protein（XBP1） and apoptosis protein C/EBP homologous protein（CHOP）， B cell lymphoma/leukemia-2（Bcl-2） and Bcl-2 associated X protein（Bax） were detected by Western blot. Deoxyribonucleotide end-transferase-mediated notch end labeling （TUNEL） was also used to detect the myocardial cell apoptosis. Result:Compared with control group， the levels of the maximum ascending rate of left ventricular pressure（+dp/dt_max）， the maximum descending rate of left ventricular pressure（-dp/dt_max） and the left ventricular systolic blood pressure（LVSP） were decreased significantly（<italic>P</italic><0.05），but the level of the left ventricular end diastolic blood pressure （LVEDP） was significantly increased（<italic>P</italic><0.05）. The expressions of GRP78， p-PERK， PERK， ATF6， XBP1， CHOP， Bax and apoptosis index were increased significantly （<italic>P</italic><0.05，<italic>P</italic><0.01）， while Bcl-2/Bax and Bcl-2 were decreased significantly（<italic>P</italic><0.01） in model group. Compared with the model group， the levels of -dp/dt_max and Bcl-2 were increased significantly （<italic>P</italic><0.05），while the level of the apoptosis index was decreased significantly（<italic>P</italic><0.01） in low-dose Wenxin granule group. In high-dose Wenxin granule group， the levels of +dp/dt_max and -dp/dt_max were increased significantly（<italic>P</italic><0.05，<italic>P</italic><0.01）， the levels of endoplasmic reticulum stress pathway protein GRP78， p-PERK， PERK， ATF6， XBP1，apoptosis-related protein CHOP， Bax and apoptosis index were decreased significantly（<italic>P</italic><0.01）， but Bcl-2 and Bcl-2/Bax were increased significantly （<italic>P</italic><0.01）. Conclusion:Wenxin granule has effect in inhibiting excessive endoplasmic reticulum stress， reducing myocardial cell apoptosis and improving cardiac hemodynamics. Its molecular mechanism may be related to decrease the levels of GRP78， PERK， p-PERK， ATF6， XBP1， CHOP， Bax and increasing the expression of Bcl-2.

Huotan Jiedu Tongluo decoction alleviates early atherosclerosis of rabbits by inhibiting eNOS uncoupling pathway / 中国中药杂志

The aim of this paper was to study the curative effect of Huotan Jiedu Tongluo (HTJDTL) decoction on a rabbit model with early atherosclerosis (AS),and furtherly to explore whether it could inhibit the BH4/eNOS uncoupling ROS or not. Twenty-four Japanese white rabbits were randomly divided into sham operation group, model group, HTJDTL decoction group and atorvastatin group. Rabbit models with early atherosclerosis were established by high fat diet, nitrogen drying and carotid artery balloon injury. The rabbits were sacrificed at 7th days after balloon injury and several parameters were measured. The pathological morphology of the common carotid artery was observed by HE staining. The blood lipids were detected by peroxidase method. The ratio of vascular eNOS dimer and monomer was measured by Western blot. The ELISA and biochemical technology were respectively used for testing BH4 and ROS levels in serum. The results showed that compared with the sham operation group, the model group had mild stenosis of the common carotid artery lumen, uneven intimal hyperplasia, lipid deposition in the intima and media, and obvious hyperplasia of the adventitia with inflammatory cell infiltration. The HTJDTL decoction could significantly inhibit the intimal hyperplasia compared with the model group, meanwhile, reduce the lipid deposition of the media and the infiltration of the adventitial cells. Compared with the sham operation group, the blood lipids and ROS of the model animals significantly increased, but BH4 and the ratio of eNOS dimer/monomer decreased. Compared with the model group, HTJDTL decoction significantly reduced the TC, ox-LDL and ROS levels, and also up-regulated eNOS dimer/monomer ratio, but it increased BH4 trend without statistical difference. According to the results, it was found that HTJDTL decoction couldsignificantly prevent and improve the vascular remodeling of rabbits model with early atherosclerosis. The mechanism of decoction may largely be related to the inhibition of BH4/eNOS uncoupling and the reduction of oxidative stress.

Combined therapy with methylprednisolone and ulinastatin in experimental autoimmune encephalomyelitis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Our previous study had demonstrated that ulinastatin (UTI) had a neuroprotective effect in experimental autoimmune encephalomyelitis (EAE). Methylprednisolone has been recommended to be a standard drug in multiple sclerosis (MS) therapies. The present study was to investigate the protective effects of UTI combined methylprednisolone in EAE.</p><p><b>METHODS</b>Mice were divided into a UTI treatment group, a methylprednisolone treatment group, a combined treatment group with UTI and methylprednisolone, a normal saline treatment group, and a normal control group. EAE mice were induced in groups receiving different combined treatments, or respective monotherapies. Demyelination was evaluated by Solochrome cyanin staining. 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP)/ myelin basic protein (MBP)/ the precursor form of nerve growth factor (proNGF)/p75/ inducible nitric oxide synthase (iNOS) proteins in cerebral cortex of EAE were detected by Western blotting.</p><p><b>RESULTS</b>The combined treatment group had a lower clinical score (0.61 ± 0.06) and demyelinating score (1.33 ± 0.33) than the groups with normal saline (clinical score: 1.39 ± 0.08, P < 0.001; demyelinating score: 2.75 ± 0.49, P < 0.05) or monotheraphies. Compared with the saline treated EAE group, UTI combined methylprednisolone significantly increased expressions of CNP (1.14 ± 0.06 vs. 0.65 ± 0.04, P < 0.001), MBP (1.28 ± 0.14 vs. 0.44 ± 0.17, P < 0.001), and decreased expressions of proNGF (1.08 ± 0.10 vs. 2.32 ± 0.12, P < 0.001), p75 (1.13 ± 0.13 vs. 2.33 ± 0.17, P < 0.001), and iNOS (1.05 ± 0.31 vs. 2.17 ± 0.13, P < 0.001) proteins in EAE. Furthermore, UTI combined methylprednisolone could significantly upregulate MBP (1.28 ± 0.14 vs. 1.01 ± 0.15, P < 0.05) expression and downregulate iNOS (1.05 ± 0.31 vs. 1.35 ± 0.14, P < 0.05) expression compared to methylprednisolone treatment EAE group. And proNGF expression was significantly lower in combined treatment (1.08 ± 0.10) than that in UTI (1.51 ± 0.24, P < 0.05) or methylprednisolone (1.31 ± 0.04, P < 0.05) treatment group.</p><p><b>CONCLUSION</b>Combination treatment of UTI with methylprednisolone was shown to protect EAE, suggesting that combination therapy is a potential novel treatment in MS.</p>

Alteration of ion channel gene expression profile in rat model of post-myocardial infarction heart failure / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the ion channel genes related with the genesis and development of heart failure after myocardial infarction through analyzing the differential gene expressions in non-infarcted myocardial tissues of post-myocardial infarction heart failure (PIHF) rat model, and that of normal rat, and the bio-informatics Stc-GO analysis on them.</p><p><b>METHODS</b>Rat model of PIHF was established by left anterior descending coronary artery ligation in six SD rats, and a control group with six sham-operative rats was set. Myocardial samples were taken in two batches from them (three rats in each group) at the heart failure formation stage and the stable stage (10 days and 8 weeks after operation) respectively. Total RNA extraction, probe preparation with reverse transcription, hybridization with double-channel cDNA microarray of rat's ion channel genes, and computerized differential gene expression screening were conducted, and Stc-GO functional clustering analysis was performed on the outcomes obtained to search out the GO sort of significance in them.</p><p><b>RESULTS</b>At 10 days after operation, 319 common differential expressed genes were found from the 746 target genes, all of them were up-regulated. At 8 weeks after operation, in the 276 differential expressed genes, 274 were up-regulated while the other two down-regulated. The up-regulated genes were those concerning receptors of various hormones, cytokines, neuro-hormones, growth factors and nuclear receptor, protein phosphorylase, G protein, various ion channels mediated by ligand or voltage, transport protein, receptor interfering protein, etc. The down-regulated genes concerning the potassium channel and transport protein, etc. Stc-GO analysis found that the six genes concerning adrenergic receptor kinase beta 1 (betaARK1), amiloride-sensitive cation channel 2 neuronal (Accn2), voltage-dependent calcium ion channel gamma subunit 1 (Cacng1), cyclic nucleotide gated channel alpha 1 (Cnga1), Glutamate receptor ionotropic kainite 2 (Grik 2) and neurotrophic tyrosine kinase receptor type 2 (Ntrk 2), were all the significantly up-regulated differential genes of the model group related with the sham-operative group, and all showed a down-regulating trend as time goes on, and four genes in them were validated by the RT-PCR test.</p><p><b>CONCLUSION</b>Ion channel genes concerning Accn2, Grik2, Ntrk2 and Cacng1 were up-regulated in PIHF, and its mechanism is waiting for further study.</p>

Characteristics of TCM syndrome and appraisal with echocardiography in heart failure rats after myocardial infarction / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the characteristics of the TCM syndrome and the changes of ventricular structure and function in heart failure (HF) rats after- myocardial infarction (MI).</p><p><b>METHODS</b>Rats were randomly divided in to the model group and the sham-operative group. The HF rat with MI model was induced by ligation of the left coronary artery. Eight weeks after operation, appraisal on TCM syndrome revealed in the model was made from the aspects of general status, breathing frequency, heart rate, exhausting swimming time and electrocardiogram, and left ventricular structure and function were observed with echocardiography.</p><p><b>RESULTS</b>Eight weeks after operation, as compared with those in the sham group, in the model group, the heart rate and breathing frequency were accelerated, the exhausting swimming time shortened, the echocardiogram parameters such as interventricular septum end-diastole thickness (IVSTd), posterior wall end-diastole thickness (PWTd), posterior wall end-systolic thickness (PWTs), ejection fraction (EF) and fractional shortening (FS) of the left ventricular reduced (P < 0.01), while left ventricular end-diastolic dimension (LVDd) and end-systolic dimension (LVDs) obviously increased (P < 0.01). In 12 leads electrocardiogram, the leads of ST segment elevated and abnormal Q wave increased. Additionally, the ratio of whole heart weight/body weight increased (P < 0.05).</p><p><b>CONCLUSION</b>The HF rats after MI manifests Xin-qi deficiency and blood stasis syndrome, and shows the pathological changes of left ventricular remodeling and function impairment.</p>