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ObjectiveTo investigate the effect of Rehmanniae Radix Praeparata on neurological function injury in ischemic stroke rats and explore its mechanism. MethodMale SD rats were randomized into sham operation， model， low- and high -dose （3.5 g·kg-1 and 7 g·kg-1） Rehmannia Radix Praeparata， and nimodipine （0.01 g·kg-1） groups. The rat model of middle cerebral artery occlusion （MCAO） was established with the modified suture occlusion method. Zea-Longa 5-point scoring was employed to evaluate the neurological function of rats. The cerebral infarction volume was detected by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Hematoxylin-eosin staining and Nissl staining were employed to observe the morphology and damage of the brain tissue. Meanwhile， the serum levels of lactate dehydrogenase （LDH）， oxidative stress-related indicators superoxide dismutase （SOD）， glutathione peroxidase 4 （GPX4）， and malondialdehyde （MDA）， and the iron （Fe） content in the brain tissue were determined. To explore the mechanism of Rehmanniae Radix Preparata in mitigating the neurological damage in ischemic stroke rats， Western blotting was employed to determine the expression levels of proteins in the ischemic brain tissue. The autophagy-associated proteins included autophagy effector （beclin-1）， microtubule-associated protein light chain 3 （LC3B）， and ubiquitin-binding protein p62 （p62）. The ferroptosis-associated proteins included transferrin （TF）， transferrin receptor 1 （TFR1）， ferritin heavy chain 1 （FTH1）， and ferropotin （FPN1）. The neurological function injury-associated proteins included brain-derived neurotrophic factor （BDNF） and tyrosine kinase receptor B （TrkB）. ResultCompared with the sham operation group， the model group showed increased neurological function score， cerebral infarction volume， and appearance of nuclear pyknosis and vacuole of cells in the cerebral cortex. In addition， the model group presented elevated levels of LDH， MDA， and Fe （P<0.01） and lowered levels of SOD and GPX4 （P<0.01）. Compared with the model group， Rehmanniae Radix Praeparata decreased the content of LDH， MDA， and Fe （P<0.05， P<0.01） and elevated the levels of SOD and GPX4 （P<0.05， P<0.01）. Compared with the sham operation group， the modeling promoted the expression of beclin-1，LC3B Ⅱ/Ⅰ， TF， and TFR1 and inhibited the expression of p62， FTH1， FPN1， BDNF， and TrkB （P<0.01）. The expression levels of these proteins were recovered after the treatment with Rehmanniae Radix Praeparata. ConclusionRehmanniae Radix Praeparata may inhibit ferroptosis and improve the neurological function in ischemic stroke rats by down-regulating the autophagy level in the brain tissue.
ABSTRACT
ObjectiveThe pulmonary edema （PE） model where the disease was located in the viscera was established according to the treatment principle that patients with the disease location inside should be treated with descending and sinking medicine， combined with changes in the disease tendency， to verify the scientificity of descending and sinking properties of Descurainiae Semen Lepidii Semen （SD）， and to preliminarily elucidate the scientific connotation of descending， ascending， floating and sinking of Chinese medicine. MethodSixty male SD rats were randomly divided into normal control group， model group （20 mg·kg-1）， positive drug group （dexamethasone， 0.075 mg·kg-1） and SD low （1.167 g·kg-1）， medium （2.334 g·kg-1）and high （4.668 g·kg-1） dose groups. The PE model was established by intrapleural injection of 1% carrageenan （2 mL·kg-1）. The evaluation indexes （lung autopsy， amount of pleural effusion， number of white blood cells， lung wet/dry weight ratio， lung water content and lung permeability） were tested to determine the optimal dose of SD decoction for intervention of PE. The relevant indexes of the five major systems （central nervous system， respiratory system， circulatory system， digestive system and urinary system） closely related to the body's Qi movement were detected and changes in the disease tendency in PE rats were analyzed， to determine the descending， ascending， floating and sinking properties of SD. In addition， histopathological changes were investigated by hematoxylin-eosin （HE） staining， and types and numbers of inflammatory cells and mediators were detected to preliminarily explore the mechanism of SD in improving PE. ResultCompared with the conditions in the normal control group， the amount of pleural effusion， number of white blood cells in pleural effusion， lung wet/dry weight ratio， lung water content and lung permeability were increased （P<0.01） in the model group， where the rats presented cough， dyspnea， shortness of breath and arched back， and a small number of them had wet nose and bubble liquid in nostrils. In the autopsy of the rats in the model group， the lungs were enlarged or accompanied by congestion and plenty of pink bubble liquid appeared at the trachea. Compared with the conditions in the model group， SD reduced the amount of pleural effusion， number of white blood cells in pleural effusion， lung coefficient， lung wet/dry weight ratio and lung water content （P<0.01）， and improved pulmonary edema symptoms such as damage， inflammation and infiltration around the lumen， thickening of the trachea， and accumulation of edema fluid， and the SD medium dose group had the best effect on the treatment of PE. In terms of respiratory system， compared with the normal control group， the model group had reduced latent time of cough and asthma （P<0.05， P<0.01） and elevated number of cough and wheezing （P<0.01）. The three SD groups had increased latent time of cough and asthma and decreased number of cough and wheezing （P<0.01）. In terms of urinary system， compared with the normal control group， the model group presented decreased urine volume. The SD low， medium and high dose groups had increased urine volume （P<0.05， P<0.01）， but they had no effect on perspiration. In terms of digestive system， compared with the conditions in the normal control group， the gastric residual rate and gastrin （GT） level were increased （P<0.05， P<0.01）， and the gastric emptying rate and small intestine transit rate were decreased （P<0.01）. The SD low dose group had elevated small intestine transit rate （P<0.01）， and the SD high and medium groups had enhanced gastric emptying rate and small intestine transit rate （P<0.01）， reduced gastric residual rate， lowered GT level to promote gastrointestinal movement and transportation （P<0.01）， and increased motilin （MTL） level to promote gastric emptying in rats （P<0.05， P<0.01）. In terms of circulatory system， compared with the normal control group， the model group displayed reduced left ventricular ejection fraction （LVEF）， left ventricular short axis shortening rate （LVFS） and cardiac output （CO） （P<0.01）， and elevated tendency of heart rate， systolic blood pressure （SBP， P<0.01） and diastolic blood pressure （DBP， P<0.05）. Compared with the model group， the SD low dose group increased LVEF and decreased DBP （P<0.05）， while the SD medium dose group increased LVEF， LVFS， CO and SBP （P<0.01） and decreased DBP （P<0.05）， and the SD high dose group increased LVFS （P<0.01） and decreased SBP （P<0.01） and DBP （P<0.05）. In terms of central nervous system， compared with the conditions in the normal control group， the standing times dropped in the model group （P<0.01）. SD reduced the movement distance， movement time， standing times and activity time in the center of the open field， and increased the rest time and activity time at the edge of the open field （P<0.05， P<0.01）. Moreover， compared with the normal control group， the model group had serious inflammatory infiltration around the lung lumen， thickened trachea with accumulated edema fluid， seriously damaged lung tissue， increased number of white blood cells and percentage of neutrophils in blood （P<0.01）， elevated percentage of monocytes， interleukin-4 （IL-4）， immunoglobulin E （IgE） level and reactive oxygen species （ROS） level in lung tissue （P<0.05，P<0.01）， and decreased IFN-γ in alveolar lavage fluid （P<0.01）. Compared with the model group， SD decreased the number of white blood cells， neutrophil accumulation， pulmonary congestion and interstitial edema， IFN-γ and IL-4 levels in alveolar lavage fluid and ROS level in lung tissue， and increased IgE level （P<0.05， P<0.01）. ConclusionSD had a significant improvement effect on PE model where the disease was located in the viscera. It could regulate the excretion of water by purgation， regulate Qi movement and expel Qi stagnation by descending and sinking lung Qi， and promote purification and descent of lung qi to make Qi movement downward. This indicated SD had the descending and sinking properties. The medium dose of SD decoction exerted the best effect， and its mechanism of action might be through regulating the neutrophil inflammatory response.
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@#Objective To investigate the features of eating difficulties in acute stroke patients, and its related factors. Methods From April to June, 2016, 192 stroke patients within 30 days were conveniently sampled, and investigated with a questionnaire about eating difficulties. Results The total rate of eating difficulties was 66.67% (including 15 nasal feeding patients). Among the 177 patients with oral feeding, the incidence of eating difficulties was 113, including 66 with feeding difficulties, 28 with dysphagia, and 91 with loss of energy and appetite, while there were 19 patients reporting all the problems above. The incidence of eating difficulties was various with the National Institutes of Health Stroke Scale (NIHSS) scores, Drinking Test scores, aphasia, prosopoplegia, stay in bed, eating assistance, activities of daily living (ADL) score, and oral health score (P<0.05). Myodynamia of upper limbs and ages also related with eating difficulties (P<0.05). The nutrition score of patients with eating difficulties was lower than those patients without eating difficulties. NIHSS score, eating assistance, ADL score, oral health score were the independent factors related with eating difficulties through multiple linear regression.Conclusion Eating difficulties are common in acute stroke patients, not only from dysphagia, but also about feeding difficulties and loss of energy and appetite. High NIHSS score, eating assistance, lower ADL score, bad oral health condition are the related factors of eating difficult, which may result in nutrition problems.
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This article was aimed to study the impact on substance and energy metabolism by chemical split fractions of Mori Cortex among hypoglycemic diabetic mouse model, in order to explain the new hypothesis of the science connotation in nature and flavor of traditional Chinese medicine (TCM). Male Kunming mice were intraperitoneally injected with a large dose of streptozotocin (STZ) (170 mg·kg-1) to establish type 1 diabetes mellitus mouse model. Medication was given consecutively for four weeks. The enzyme-linked immunesorbent assay (ELISA) was used to detect glucosekinase (GCK), glycogen phosphorylase (PYGL), pyruvate dehydrogenase (PDH), α-ketoglutarate dehydrogenase (α-KGDHC), phosphoglycerate kinase (PGK), acetyl coenzyme A (acetyl-CoA), adenylate kinase (ADK), fumarase (FUM), cytochrome C reductase (CCR), cytochrome C oxidase (COX) and other indicators. Enzymatic detection was used to determine the content of ATP coenzyme (ATPs), the content and ratio of NAD and NADH, the content of myocardial cell Na+-K+ ATP enzyme, as well as the content of ATP and ADP. The results showed that in the model group, the expression of PYGL was increased; and the expressions of GCK and PDH were decreased. It prompted that the source of glucose increased and the expelling of glucose decreased. The glucose level was increased. The COX expression was reduced and the respiratory chain was blocked. It regulated oxidative phosphorylation and the substrate phosphorylation level. It upregulated the expression of CCR, ATPs, NAD+, PGK, α-KGDHC and ADK. However, the expression of FUM was decreased. The activity of Na+-K+ ATPase was decreased significantly. At last, the metabolic disorders appeared. Mori Cortex aqueous extracts and the chemical split fractions significantly increased the GCK and PDH level in substance metabolism among diabetic mice. The levels of PYGL, α-KGDHC, PGK and acetyl-CoA were decreased (P < 0.05, or P <0.01). Meanwhile, it increased ATP and FUM, myocardial cell Na+-K+ ATP enzyme, and COX level in the energy metabolism (P < 0.05). It decreased the level of NAD+, CCR and ATPs (P < 0.05, or P <0.01). It was concluded that both the aqueous extracts and chemical split fractions of Mori Cortex can effectively improve the substance and energy metabolism disorders of diabetic mouse model. This effect may be related to the cold nature of Mori Cortex.
ABSTRACT
This article was aimed to study effects of aqueous extracts and all chemical split fractions ofMori Cortex on hypoglycemic effect of diabetic mice model. Intra-peritoneal injection of 170 mg·kg-1 streptozocin (STZ) was given to male Kunming mice to establish type I diabetes mode. Continuous administration of medication was given for 4 weeks. And then, indicators such as body weight, water intake, food intake, fasting plasma glucose (FBG), insulin, C-peptide, total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-c), and low density lipoprotein cholesterol (LDL-c) were detected. Pathological morphology of the liver and pancreas were observed by light microscopy. The results showed that high-dose group ofM. Cortex aqueous extracts can improve weight loss of type I diabetic mice, significantly reduce water intake and food intake, reduce FBG, TC, TG and LDL-c levels in different degrees (P<0.05 orP<0.01), and increase C-peptide and HDL-c levels (P<0.05 orP<0.01). When dosages of 30% ethanol fraction and fatty oil fraction were only about 1/2 and 1/4 ofM.Cortex aqueous extracts, we found that it can improve lipid disorder status,repair liver cells, and improve liver tissue damage. Its effect was superior to M. Cortex aqueous extracts. It was concluded thatM. Cortexaqueous extracts showed a better hypoglycemic effect. The effective component parts were 30% ethanol fraction and fatty oil fraction. Its hypoglycemic mechanism may be related to the promotion of insulin secretion, regulation of blood lipid disorders, as well as the protection of liver structure and function.
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This study was aimed to investigate the antitussive, expectorant and antiasthmatic effects of aqueous extracts and the chemical split fractions ofMori Cortex. Cough mice models induced by ammonia water were used to observe the effect on arresting cough. The phenol red expectoration method in mice was used to observe the effect on expelling phlegm. Histamine and acetylcholine mixture induced asthma model was used to observe the effect on relieving asthma. Isolated trachea model was used to observe the effect on relieving spasm. Compared with the control group, the low, medium and high doses of aqueous extracts ofM. Cortex can obviously decrease the frequency of cough, increase phenol red output of trachea in mice, prolong the latent period of asthma in guinea pigs, and increase the antispasmodic rate. The medium dose had the best effect. The comparison between different chemical split fractions ofM. Cortex and the control group showed that the 30% and 50% ethanol fraction ofM. Cortex can obviously decrease the frequency of cough and prolong the latent period of cough induced by ammonia water in mice, increase phenol red output of trachea in mice (P<0.05 orP<0.01); and 30% ethanol fraction and fatty oil fraction can prolong the latent period of asthma in guinea pigs (P< 0.05 orP<0.01). In addition, 30% ethanol fraction can obviously reduce the degree of tracheal contraction. It had certain effect of relieving spasm. It was concluded that aqueous extracts ofM. Cortex had better effects on relieving cough, expectorant and antiasthma. The effective part was the 30% ethanol fraction, which was the dose equivalent of 1/3 of the medium dose. It had significant effect on relieving cough, expectorant and antiasthma. The effect was equivalent to the medium dose of aqueous extracts of M. Cortex.
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This article was aimed to study immunomodulatory effect of chemical split fractions ofMori Cortex, in order to initially explain effective parts that played a role in immunomodulatory effect ofMori Cortex. The carbon clearance test, serum hemolysin test, E-rosette test, and lymphocyte transformation test were carried out to explore influence of these chemical split fractions ofMori Cortex on immune organs, nonspecific immunity, humoral immunity and cellular immunity. The results showed that in the carbon clearance test, 50% ethanol fraction markedly reduced the thymus index (P<0.01) and the correction indexα (P<0.05). In hemolysin test, the half value hemolysis (HC50) was improved by 30% ethanol fraction and fatty oil fraction (P<0.05). Besides, in the E-rosette test, the E-rosette ration was increased in the 30% ethanol fraction group (P<0.05). In the lymphocyte transformation test, the 30% ethanol fraction can promote the thymus and spleen lymphocytes proliferation (P<0.05 orP<0.01), while the 50% ethanol fraction inhibited the proliferation (P<0.05 orP<0.01). It was concluded that the 30% ethanol fraction can boost both the humoral immunity and cellular immunity; the 50% ethanol fraction can induce the growth of thymus with a suppressive effect on nonspecific immunity and cellular immunity; the fatty oil fraction can improve humoral immunity.
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This study was aimed to investigate the estrogenic effects ofCoicis Semen in order to preliminarily discuss the mechanism. Mouse uterine weight test and MCF-7 cell proliferation assay were used to evaluate the estrogenic effects ofC. Semen. Reporter gene assays were adopted to explore the action mechanism ofC. Semen. In reporter gene assay, HEK293 cells were co-transfected with pERE-TAL-luc, pβgal-Control, pCXN2-hERα, or pCXN2-hERβ. And the expression of reporter gene luc was controlled by ERE. Mouse uterine weight test showed that compared with the control group, the aqueous extracts ofC. Semen can increase the uterus index of premature female mice (P<0.01). It can significantly promote the proliferation of MCF-7 cells in the medium without estrogen (P<0.01). The reporter gene controlled by ERE technology showed thatwhen mediated by ERα or ERβ respectively, the normalized luciferase activity of aqueous extracts ofC. Semen was significantly higher than activity of the control group (P< 0.05 orP< 0.01). It was concluded that the aqueous extracts ofC.Semen can increase the uterus index of premature female mice and promote the proliferation of MCF-7 cells in the medium without estrogen. We found the estrogenic effects ofC. Semen for the first time. And the estrogenic effects ofC. Semen were mainly mediated by ERβ.