Model Group Levels Research Articles

MiR-497 Prevents Septic Kidney Injury and Improves Immune Function Through Inhibition of Janus Kinase/Signal Transducers and Activators of Transcription (JAK/STAT) Pathway

Abnormal expression of miR-497 is related to the progression of septic renal injury. This study aimed to identify the protective effect of miR-497 on septic renal injury and immune function. We established a rat model of septic renal injury with sham-operated group and treated rats with culture solution of miR-497, gentianella acuta (positive control group), miR-497 plus JAK/STAT pathway agonist, and distilled water (model group). After treatment, urine output and renal histopathological changes were detected. Flow cytometry and RT-qPCR determined the levels of serum Scr, BUN, KIM-1, NGAL, IgG, IgA, and IgM, and evaluated the CD4+T, CD8+T, NK cell activity. Western blot assessed the activity of JAK/STAT signaling pathway. The model group and pathway agonist group had the highest 24-hour urine output, serum Scr, BUN, KIM-1, and NGAL levels, followed by miR-497 group and positive control group, and sham-operated group. The CD4+T, and NK cell activity was reduced with a drop in IgG, IgA, IgM levels in rats with septic renal injury, but treatment with miR-497 or gentianella acuta restored the cell activity and Ig levels and addition of JAK/STAT pathway agonist would further decrease the immune cell activity. There was no difference between the model group and pathway agonist group, miR-497 group and positive control group (p < 0.05). Following miR-497 group, and sham-operated group, the expression of miR-497 in model group, pathway agonist group, and positive control group was lowest (p < 0.05). As the expression of JAK and STAT did not vary among five groups (p > 0.05), we found highest expression of p-JAK and p-STAT levels in model group, positive control group, and pathway agonist group, and lowest expression in sham-operated group. miR-497 inhibits the activity of JAK/STAT signaling, up-regulates the activity of CD4+T, NK cells and levels of IgG, IgA, IgM, while inhibiting CD8+T cells activity to improve immune function. In conclusion, miR-497 attenuates septic kidney injury through inhibiting KIM-1 and NGAL expression, indicating that miR-497 and JAK/STAT pathways may be potential therapeutic targets for treating septic kidney injury.

MiR-181a regulates mitophagy and improves cognitive function in vascular dementia via targeting PINK1/Parkin pathway

This study investigates the mechanism of miR-181a in cognitive function and mitophagy in vascular dementia. A rat model of vascular dementia was established by common carotid artery ligation and then divided into model group (n= 10), mimics group (n= 10), inhibitors group (n= 10), NC group (n= 10), and the remaining 10 rats were in sham operation group. The rats in mimics group were intravenously injected with miR-181a mimics and rats in inhibitors group were injected with miR-181a inhibitors. Model group and mimics group rats had a longer time on the 3rd to 4th day than sham operation group, and inhibitors group had a shorter time than model group and mimics group (allP<0.05). Time in quadrant of the cylindrical platform in model group and mimics group was shortened while it was prolonged in inhibitors group (P< 0.05). miR-181a mRNA level in model group, inhibitors group and NC group was lower. On the contrary, PINK1 and Parkin level was higher. miR-181a mRNA level was higher in mimics group with lower levels of PINK1 and Parkin (P<0.05). miR-181a level in mimics group was significantly increased and PINK1 and Parkin mRNA was decreased. miR-181a mRNA level in inhibitors group decreased significantly, while PINK1 and Parkin mRNA was increased (P<0.05). Parkin expression in mimics group and NC group was reduced. Up-regulation of miR-181 (mimics group) can target and inhibit PINK1/Parkin mRNA, thereby inhibiting mitophagy. miR-181a downregulation in mitochondria can promote PINK1/Parkin expression, thereby activating mitophagy in rats with vascular dementia, alleviating mitochondrial dysfunction, and improving the cognitive ability of rats.

Effect of aerobic exercise on AKT/GSK3β mediated hepatocyte apoptosis in nonalcoholic fatty liver disease

To investigate the effect of aerobic exercise on AKT/GSK3β pathway-mediated hepatocyte apoptosis in non-alcoholic fatty liver diseases(NAFLD). A total of 30 6-week-old male C57BL/6J mice, and mice were fed adaptively for one week. The control group was fed with ordinary diet, and the model group and model exercise group were fed with high-fat diet until 18 weeks. At the 10th week of the experiment, the model exercise group received aerobic exercise intervention for 8 consecutive weeks until the end of the experiment at the 18th week. Automatic biochemical analyzer to detect serum total cholesterol(TC), triglycerides(TG), alanine aminotransferase(ALT), aspartate aminotransferase(AST), low-density lipoprotein(LDL-C) and high-density lipoprotein(HDL-C) levels. Liver pathological morphology was observed by staining with oil red O and HE. The expression changes of AKT, P-AKT~( Ser473), GSK3β, P-GSK3β~(Ser9) and Caspase-3 proteins were detected by western blot, and the apoptosis of hepatocytes was detected by in situ terminal transferase labeling(TUNEL). (1) After intervention, compared with control group, body weight, liver index, serum TC, TG, ALT, AST and LDL-C levels in model group were significantly increased(P<0.01 or P<0.05), while HDL-C level was significantly decreased(P<0.01). Compared with model group, body weight, liver index, serum TC, TG, ALT, AST and LDL-C levels in model exercise group were significantly decreased(P<0.01 or P<0.05), while HDL-C level was significantly increased(P<0.01). (2) Compared with the control group, hepatocyte steatosis and the number of lipid droplets in model group were significantly increased. Compared with the model group, the degree of hepatic adipose degeneration was significantly improved and the number of hepatic lipid droplets was significantly decreased in the model exercise group. (3) Compared with control group, the protein expression levels of P-AKT~(Ser473) and P-GSK3β~(Ser9) in model group were significantly decreased(P<0.01 or P<0.05), the protein expression levels of Caspase-3 were significantly increased(P<0.05), and the number of hepatocyte apoptosis was significantly increased(P<0.05). Compared with model group, the expression of P-AKT~(Ser473) and P-GSK3β~(Ser9) protein in model exercise group was significantly increased(P<0.01 or P<0.05), the expression of Caspase-3 protein was significantly decreased(P<0.05), and the number of hepatocyte apoptosis was significantly decreased(P<0.01). Aerobic exercise can effectively improve NAFLD, by activating AKT/GSK3β pathway and increasing the expression of AKT/GSK3β pathway related molecules, thereby reducing caspase-3 expression and hepatocyte apoptosis.

Protective Role of Tripterygium wilfordii Polyglycosides in Rheumatoid Arthritis by Regulating Janus Kinase 2/Signal Transducer and Activator of Transcription 3 Pathway

To investigate the effect and mechanism of Tripterygium wilfordii polyglycosides on rheumatoid arthritis rats by regulating the signal transducer and activator of transcription 3 pathways. 45 specific-pathogen free male Sprague-Dawley rats were randomly divided into normal control group, model group and Tripterygium wilfordii polyglycosides group with 15 rats in each group. The histopathological changes of the synovium of the ankle joint of rats in each group, on the 12th, 20th and 28th d were compared. On the 12th d, the degree of tissue swelling in the model group was significantly higher than that in the normal control group (p<0.05); there was no significant difference in the degree of toe swelling between the Tripterygium wilfordii polyglycosides group and the model group (p>0.05). At 20 d and 28 d, the degree of toe swelling in the model group was significantly higher than that in the normal control group (p<0.05); the swelling degree of toes in Tripterygium wilfordii polyglycosides group was significantly lower than that in model group (p<0.05). The serum interleukin-6 level in model group was significantly higher than that in normal control group and interleukin-10 level was significantly lower than that in normal control group (p<0.05). The level of interleukin-6 in serum of rats in Tripterygium wilfordii polyglycosides group was significantly lower than that in model group and the level of interleukin-10 was significantly higher than that in model group (p<0.05). The expression levels of phosphorylated Janus kinase 2 and phosphorylated signal transducer and activator of transcription 3 protein in model group were significantly higher than those in normal control group; the expression levels of phosphorylated Janus kinase 2 and phosphorylated signal transducer and activator of transcription 3 protein in Tripterygium wilfordii polyglycosides group were significantly lower than those in the model group (p<0.05). Tripterygium wilfordii polyglycosides has the effect of anti-rheumatoid arthritis and its mechanism may be through regulating the Janus kinase 2/ signal transducer and activator of transcription 3 signal pathways by reducing the expression level of phosphorylated Janus kinase 2 and phosphorylated signal transducer and activator of transcription 3 proteins.

Effect of berberine on hyperandrogenemia, ovulation dysfunction and inflammation in a mouse model of polycystic ovary syndrome

Purpose: To study the effect of berberine (BBR) on hyperandrogenemia (HA), ovulation dysfunction and inflammation in a mouse model of polycystic ovary syndrome (PCOS).Methods: Forty-five female Kunming mice were randomly divided into control group (9 mice), and mice injected with dehydroepiandrosterone (n = 36) for establishment of PCOS model. The PCOS mice were randomly divided into model group, low-dose BBR (0.25 g/kg), medium-dose BBR (0.5 g/kg) and highdose (1.0 g/kg) groups, with 9 mice in each group. Changes in ovarian morphology were monitored, and sex hormone levels i.e. testosterone (T) and luteinizing hormone (LH)), and inflammatory factors were determined.Results: The model group levels of T and LH were significantly higher than those of the blank control group (p &lt; 0.05), but T and LH levels were significantly lower in middle- and high-dose BBR groups than in the model mice. There were marked increases in IL-6 and TNF-α levels in model mice, when compared to blank control mice, but reduced in the mice treated at the 3 doses of BBR, relative to model mice (p &lt; 0.05). In contrast, the number of follicles was higher at each stage of development in mice for each BBR dose than in the model mice, with increase in corpus luteum.Conclusion: Berberine lowers the weight of PCOS mice, mitigates hyperandrogenemia and inflammatory state, and enhances recovery of ovulation. However, there is need for further studies on its clinic applicability.&#x0D; Keywords: Berberine, Polycystic ovary syndrome, Hyperandrogenemia, Ovulation dysfunction, Inflammatory

MiR-23a-5p exacerbates intestinal ischemia–reperfusion injury by promoting oxidative stress via targeting PPAR alpha

MiR-23a-5p is involved in the occurrence and development of some serious diseases, but its effects on intestinal ischemia–reperfusion (II/R) injury is unclear. In this research, the hypoxia/reoxygenation (H/R) model on IEC-6 cells and II/R model in mice were used. The data showed that the ROS level in model group was significantly increased compared with control group. The level of intestinal MPO was increased and serum SOD was decreased in mice compared with sham group. Moreover, the expression levels of miR-23a-5p in model groups were obviously increased in vitro and in vivo, while the expression levels of PPARα, FOXO3α, PGC-1α, Nrf2, CAT, NQO1, HO-1 and SOD2 were significantly decreased. The double luciferase reporter gene assay showed that there was binding site between miR-23a-5p and PPARα. When miR-23a-5p was inhibited or PPARα gene was overexpressed, H/R-caused cell damage was alleviated and ROS level was decreased compared with NC group. PPARα expression level was increased, accompanied by the increased levels of FOXO3α, PGC-1α, Nrf2, CAT, NQO1, HO-1 and SOD2. After enhancing miR-23a-5p expression or silencing PPARα gene, H/R-caused cell damage was further aggravated compared with NC group, and ROS level was increased associated with the decreased levels of FOXO3α, PGC-1α, Nrf2, CAT, NQO1, HO-1 and SOD2. Our study demonstrated that miR-23a-5p exacerbated II/R injury by promoting oxidative stress via targeting PPARα, which should be considered as one new drug target to treat II/R injury.

LncRNA-mRNA competing endogenous RNA network in IR-hepG2 cells ameliorated by APBBR decreasing ROS levels: a systematic analysis.

BackgroundRadix Astragali (Astragalus membranaceus var. mongholicus (Bunge)) and Coptis chinensis (Coptis chinensis var. angustiloba) are two commonly prescribed traditional Chinese herbs for diabetes. Astragalus Polysaccharide (AP) and Berberine (BBR) are active ingredients of these two herbs respectively and they are scientifically proved to have immunomodulatory and anti-inflammatory effects. They are also known for their antidiabetic potential by ameliorating insulin resistance (IR). AP and BBR have shown different advantages in treating diabetes according to previous reports. However, very few studies focus on the combined activities of the two potential antidiabetic ingredients. In this study, we discovered that reactive oxygen species (ROS) accumulated in IR-hepG2 cells and APBBR can decrease ROS level in model group significantly. We conjectured that APBBR can ameliorate IR in hepG2 cells by decreasing ROS level. In order to verify this hypothesis, we obtained phenotype and transcriptome information of IR-HepG2 cells and explore the underlying mechanism of the combination of AP and BBR(APBBR) activity on the relationship between ROS change in IR at whole-transcriptome level, so as to shed new light to efficacy and application of APBBR in treating diabetes.MethodsThe IR cell model was established with high-level insulin intervention. Glucose content, HepG2 cell viability as well as ROS level was detected to study the effect of IR-hepG2 cell phenotype. Unbiased genome-wide RNA sequencing was used to investigate alterations in experimental groups. Then, GO and KEGG functional enrichment was performed to explore the function and pathway of target genes. Venn analysis found out the differentially expressed lncRNAs that had close relationship with IR and ROS. Finally, we screened out candidate lncRNAs and these target genes to construct interaction network of differentiated lncRNA–miRNA–mRNA by according to the principle of competitive endogenous RNA (ceRNA).ResultsThe biochemical experiments showed that APBBR administration could improve the proliferation activity of IR-HepG2 cells and decrease ROS level in model cells. The GO and KEGG functional enrichment analyses demonstrated several mRNAs remarkably enriched in biological processes and signaling pathways related to ROS production and IR progression. Interaction network suggest that APBBR ameliorates IR in HepG2 cells by regulating the expression of multiple genes and activating relevant signaling pathway to decrease ROS level. Thus, we demonstrated that APBBR ameliorated IR in hepG2 cells via the ROS-dependent pathway.

Effect of Ellagic Acid on Wound Healing of Chronic Skin Ulceration in Diabetic Mice and Macrophage Phenotype Transformation

Objection: To study the effect of modified formulae for ellagic acid (EA) on wound healing and macrophage phenotype transformation in diabetic mice. Materials and Methods: 10 db/m mice regarded as normal group, Dividing 20 db/db mice as the model group and EA group, 10 in every group. Surgical operation on the back was made to prepare experimental wound models. External medicines were applied and changed dressings every other day and measurement. Wound tissues were gained when animals were sacrificed by anesthesia 7 days later. Pathological changes were observed by HE and Masson staining methods, while using RT-PCR and IHC to measure mRNA and protein levels of MI/M2 type cytokines of the wound tissues. Results: Compared with the normal group, the wound area of db/db was greater (P &lt; 0.001), while EA group less (P &lt; 0.001). IL-12 and TNF-α mRNA and proteins levels in model group were higher (P &lt; 0.001, respectively), while VEGF and TGF-βmRNA and proteins were lower. mRNA and protein levels of inflammato cytokines of EA group was all lower (P &lt; 0.001), while the growth factor were significantly higher (P &lt; 0.001). Conclusion: Wound healing of db/db mice was delayed and modified formulae for EA had effects on wound healing. The way by decreasing the Ml type inf1ammatory factors and improving the M2 type growth factors might be one of the mechanisms for wound healing.

Effects of ivabradine hydrochloride combined with trimetazidine on myocardial fibrosis in rats with chronic heart failure.

Effects of ivabradine hydrochloride (Iva) and trimetazidine on myocardial fibrosis (MF) in rats with chronic heart failure (CHF) werφe explored. Fifty Wistar rats were randomly divided into sham operation, model, Iva, trimetazidine and combined drug group with 10 rats each. All rats except those in sham operation group were subjected to establish CHF model by constricting the abdominal aorta. After successful modeling, rats in the sham operation and model group received normal saline (10 mg/kg) gavage daily, the Iva group received Iva (10 mg/kg) gavage, the trimetazidine group received trimetazidine (10 mg/kg) gavage, and the combined drug group were given Iva (10 mg/kg) and trimetazidine (10 mg/kg) gavage for 12 weeks. The changes of hemodynamic indexes and heart rate, connective tissue growth factor (CTGF) and superoxide dismutase (SOD) levels as well as transforming growth factor β1 (TGF-β1) and collagen I (COL-I) expression levels in myocardial tissue of each group were detected. Compared with sham operation group, the left ventricular end-diastolic pressure (LVEDP) level, CTGF expression, TGF-β1 mRNA and COL-I mRNA expression levels in model group increased significantly, but the ± dp/dtmax and SOD content in myocardial tissue decreased significantly. Compared with model group, the LVEDP level, CTGF expression, TGF-β1 mRNA and COL-I mRNA expression levels in Iva group, trimetazidine group and combined drugs decreased significantly, but the ± dp/dtmax and the SOD content in myocardial tissue increased significantly (P<0.05). Changes in the combined drug group were the most notable (P<0.05). Iva combined with trimetazidine reduces LVEDP in rat with CHF, increases SOD content, and inhibits CTGF expression and TGF-β1 and COL-I expression levels in myocardial tissues, thus achieving the inhibitory effect on MF.

Effects of hypoxia ischemia on caspase-3 expression and neuronal apoptosis in the brain of neonatal mice.

Effects of hypoxia ischemia on caspase-3 expression and neuronal apoptosis in the brain of neonatal mice were investigated. Twenty-five neonatal CD1 mice aged 1 week were selected and randomly divided into sham-operation group (n=8) and newborn hypoxia ischemia encephalopathy (NHIE) model group (n=17). The messenger ribonucleic acid (mRNA) expression levels of caspase-3 and Fas ligand (FasL) in brain tissues of mice in both groups were detected via reverse transcription-polymerase chain reaction (RT-PCR). The protein expression levels of caspase-3 and FasL in mice in both groups were detected via western blotting. Moreover, apoptosis of brain tissues was detected using the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), and caspase-3 protein expression level in brain tissues was detected using immunohistochemical methods. Results of RT-PCR and western blotting revealed that compared with those in sham-operation group, caspase-3 and FasL expression levels in model group were significantly increased. Results of TUNEL showed that the number of apoptotic neurons in model group was significantly increased. Besides, results of immunohistochemical detection manifested that the caspase-3 protein expression level in model group was obviously increased. Hypoxia ischemia can lead to significant increase of caspase-3 expression and increase of neuronal apoptosis in the brain of neonatal mice.

Changes of helper T cell 17/regulatory T cells and cytokines in peripheral blood of rats with acute alcoholic liver injury and clinical significance

Objective To investigate the changes of helper T cell 17 (Th17)/regulatory T cell (Treg) cells and cytokines in peripheral blood of rats with acute alcoholic liver injury and the clinical significance. Methods Thirty-six male SD rats were randomly divided into control group and model group. The animals in the model group were given 30°, 40° or 50° liquor at a dose of 10 ml/kg for 2 consecutive weeks, and those in the control group were given an equal volume of distilled water. The aspartate aminotransferase (AST), alanine aminotransferase (ALT) and total bilirubin (TBIL) levels in peripheral blood were determined using AST, ALT and TBIL test kits. The liver function damage was evaluated by hematoxylin and eosin (HE) staining. The expression of Th17 and Treg cells were analyzed by flow cytometry. The serum levels of interleukin (IL)-6, IL-17, tumor necrosis factor-α (TNF-α), IL-10 and IL-35 were measured by enzyme linked immunosorbent assay (ELISA). Results As compared with the levels of AST [(130.66±12.07) U/L], ALT [(60.49±6.20) U/L] and TBIL [(30.83±3.22) μmol/L] in control group, the AST [(195.61±20.74) U/L], ALT [(88.16±9.33) U/L] and TBIL (48.54±5.41) μmol/L levels in model group were significantly increased (t=3.419, 3.174 and 4.058 respectively, P<0.05). Pathological staining showed that the liver tissue of the model group was seriously damaged. As compared with the proportion of Th17 [(2.48±0.30)%], Treg [(7.74±0.79)%] and the Th17/Treg ratio (0.33±0.07) in control group, the proportion of Th17 [(5.87± 0.61)%] and Th17/Treg ratio (1.76±0.22) in model group was significantly increased [(5.87± 0.61)%], and the proportion of Treg cells was significantly decreased [(3.25±0.35)%, t=4.331, 4.428 and 5.663 respectively, P<0.01]. As compared with the expression of IL-6 [(28.06±3.03) ng/L], IL-17 [(21.39±2.63) ng/L], TNF-α [(70.56±6.99) ng/L], IL-10 [(25.67±2.42) ng/L] and IL-35 [(33.17±3.25) ng/L] in control group, the levels of IL-6 [(62.81±6.55) ng/L], IL-17 [(52.46±5.40) ng/L] and TNF-α [(161.17±17.32) ng/L] in model group were significantly increased in model group (t=4.220, 4.316, 4.207 and 3.574 respectively, P<0.05), and those of IL-10 [(15.72±1.70) ng/L] and IL-35 [(19.66±2.03) ng/L] were significantly reduced (t=3.618, P<0.05). Conclusion The imbalance of Th17/Treg cells and unusual expression of cytokines may play an important role in acute alcoholic liver injury. Key words: Acute alcoholic liver injury; Lymphocyte subset; Cytokines

San-Ao Decoction () Regulates Urine Volume on Bronchial Asthma Model Mice.

To observe the effect of San-Ao Decoction (, SAD) on water metabolism of bronchial asthra model mice. Forty-five female BALB/c mice were randomly divided into control, model and SAD groups by a random number table, 15 mice in each group. A composite method with ovalbumin (OVA) sensitization and challenge was developed to establish bronchial asthma model. Mice in the control group were intraperitoneally injected with distilled water without aerosol inhalation challenge. On day 15-22, 0.3 mL SAD was administered via gastric route in SAD group, one time per day, while an equivalent volume of normal saline was used for gastric administration in the control and model groups. Changes in airway resistance in the inspiratory phase (RI-R-Area) were detected using an AniRes2005 system, and 5-h urine output was collected by metabolic cages. Histopathological changes in lung and kidney were observed by hematoxylin-eosin staining. mRNA expressions of aquaporin (AQP) 1 and AQP2 in kidney were detected by reverse transcription-polymerase chain reaction, and the protein expressions of AQP1 and AQP2 in kidney were detected by immunohistochemistry. Enzyme-linked immune sorbent assay was used to detect the OVA-specific endothelium-1 (ET-1), antidiuretic hormone (ADH), atrial natriuretic peptide (ANP), prostaglandin E2 (PGE2), and angiotensin II (Ang II) levels in serum, lung and kidney tissues, respectively. The nitric oxide (NO) contents in serum, lung, and kidney tissues were tested by chemical method, respectively. Compared with the control group, the serum IgE level in model group increased (P<0.01). Following the pathologic changes in lung tissue, no significant change in kidney tissue was observed among 3 groups. Compared with the control group, the mice in the model group showed elevated airway resistance during inhalation phase, higher mRNA and protein expression levels on AQP1 and AQP2 in kidney tissue and higher ET-1 levels in serum, lung and kidney tissues, ADH and ANP in lung and serum, PGE2 in kidney, Ang II in lung and kidney tissues (P<0.05 or P<0.01), but decreased in 5-h urinary output as well as NO and PGE2 contents in serum and lung tissues (P<0.05 or P<0.01). Compared with the model group, the mice in the SAD group showed a weakened airway resistance in inspiratory phase, lower mRNA and protein expressions of AQP1 and AQP2 in kidney tissues, lower levels of ET-1, ADH, ANP in serum as well as ET-1, ANP, Ang II levels in kidney tissues (P<0.05 or P<0.01), whereas 5-h urinary output, NO content in kidney, ADH, ANP and PGE2 levels in lung and Ang II in serum increased (P<0.05 or P<0.01). San-Ao Decoction can regulate the urine volume through regulating AQP1 and AQP2 expression, and the expression of these in the kidneys might be regulated by ET-1, NO and Ang II.

Dose and timing of normal saline resuscitation on endothelial glycocalyx in early septic shock

To observe the effect of different doses and timing of normal saline (NS) resuscitation combined with norepinephrine (NE) on endothelial glycocalyx in rabbits with early septic shock. Thirty New Zealand male rabbits were randomly divided into sham group, model group, 30 mL and 60 mL timely resuscitation groups (30 mL and 60 mL timely group), and 30 mL delayed resuscitation group (30 mL delayed group) with 6 rabbits in each group. The rabbit model of septic shock was reproduced by cecal ligation and puncture (CLP). The rabbits in sham group were only received abdominal cavity open without cecal and ligation. The rabbits in 30 mL and 60 mL timely groups and 30 mL delayed group were intravenous infused with 30 mL/kg or 60 mL/kg NS immediately or 1 hour after model reproduction for 1 hour, and the mean arterial pressure (MAP) was maintained over 75 mmHg (1 mmHg = 0.133 kPa) compared with intravenous pumping of 0.02-0.05 μg×kg-1×min-1 NE followed by 5 mL/h NS infusion till the end of the experiment. The rabbits in sham and model groups were only given 5 mL/h NS. The changes in arterial blood gas before and immediately after resuscitation were observed in three fluid resuscitation groups. The internal jugular vein blood was collected at 0, 3, 6 hours after model reproduction. The levels of syndecan-1 (polysaccharide envelope marker) in plasma were determined by enzyme linked immunosorbent assay (ELISA). The rabbits were sacrificed at 6 hours after model reproduction, and the lung tissue was harvested. Western Blot was used to determine the protein expressions of intercellular adhesion molecule-1 (ICAM-1), matrix metalloproteinase 2 (MMP-2) and syndecan-1. The positive expression of syndecan-1 in lung tissue was observed by immunohistochemical method. (1) Blood gas analysis: compared with the results before resuscitation, the levels of lactic acid (Lac) after resuscitation in three fluid resuscitation groups were significantly decreased, especially in 30 mL timely group; the central venous blood oxygen saturation (ScvO2) was significantly increased, especially in 30 mL delayed group. Oxygenation index (PaO2/FiO2) was improved in 30 mL timely and 30 mL delayed resuscitation groups, which was decreased in 60 mL delayed group. (2) Plasma marker: compared with sham group, plasma syndecan-1 level in model group was significantly increased with a time-dependent manner. Plasma syndecan-1 levels at 3 hours in 30 mL timely and 30 mL delayed groups were significantly decreased as compared with those of model group (ng/L: 138.0±2.4, 139.7±15.7 vs. 161.5±4.1, both P < 0.05), but it was significantly increased at 6 hours in 30 mL delayed group (ng/L: 213.1±19.4 vs. 206.4±15.5, P < 0.05). The plasma syndecan-1 levels at 3 hours and 6 hours in 60 mL timely group were significantly higher than those in model group (ng/L: 233.0±28.9 vs. 161.5±4.1, 252.3±27.2 vs. 206.4±15.5, both P < 0.05). (3) Protein expression in lung tissue: compared with sham group, the protein expressions of ICAM-1 and MMP-2 in lung tissue of model group were significantly increased, and syndecan-1 protein expression was significantly decreased. After 30 mL timely or 30 mL delayed resuscitation, the protein expressions of ICAM-1 and MMP-2 in lung tissue were significantly decreased, and syndecan-1 protein expression was significantly increased, especially in 30 mL timely group, which showed statistical differences as compared with those of model group (ICAM-1 protein: 0.56±0.09 vs. 1.04±0.05, MMP-2 protein: 0.83±0.15 vs. 1.06±0.06, syndecan-1 protein: 2.09±0.08 vs. 0.99±0.03, all P < 0.05). The change tendency of protein expressions in 60 mL timely group was opposite to the other two resuscitation groups. (4) Immunohistochemistry: the positive expression of syndecan-1 in lung tissues was significant in the sham group, and it was lowered in model group. The positive expression of syndecan-1 was increased after 30 mL timely or 30 mL delayed resuscitation, but further weakened in 60 mL timely group. The dose and timing of resuscitation with NS in septic shock can affect pulmonary vascular endothelial glycocalyx function. The timely resuscitation with 30 mL NS in combination with NE plays a protective effect on endothelial cell and glycocalyx. However, NS resuscitation which was not timely or excessive infusion can make the glycocalyx degradation more obvious, resulting in increased endothelial permeability, microcirculation damaged, thus aggravate lung injury.

Study on the effect of moxibustion at “Shénquè” ( CV 8) on the immune system of rats taking long-term exhaustive exercise

ObjectiveTo explore the effect of moxibustion at “Shénquè” (▪ CV 8) on the immune system of rats taking long-term exhaustive exercise. MethodThirty male SD rats were randomly divided into blank group, model group and moxibustion group with 10 rats in each group. Except the rats in blank group, all the rats were established into long-term exhaustive models through swimming again and again. Mild moxibustion at CV 8 for 15 min was carried out in the rats in moxibustion group after modeling. The treatment was conducted once every other day, and treatment for 10 times was needed. The exhaustive duration for each time was recorded, the levels of serum IgA, IgG, IgM and ACP were detected 24 h after the 10th exhaustive exercise, and the histomorphological changes of spleen were observed. Immunoturbidimetry was adopted for detecting the IgA level, and ELISA method was used for detecting the levels of IgG, IgM and ACP. ResultsThe common exhaustive duration in moxibustion group was longer than that in model group, however, the time difference was statistically significant after the 5th exhaustive exercise (P<0.01). The levels of serum IgG and IgM in model group were lower than that in blank group (all P<0.01), and the serum ACP level in model group was higher than that in blank group (P<0.01), indicating that the splenic histomorphology was damaged. The levels of serum IgG and IgM in moxibustion group were higher than that in model group (all P<0.01), and the serum ACP level in moxibustion group was lower than that in model group, showing that the splenic histomorphology was repaired. ConclusionMoxibustion at CV 8 can improve the levels of immunoglobulin (IgG, IgA, IgM) of tired body for a long time, decrease the ACP level, and repair the damaged spleen tissues.

Effects of puerarin on blood lipids and inflammatory factors in rats with lower limb arteriosclerosis obliterans

Abstract This study investigated the effects of puerarin (PUE) on blood lipid and inflammatory factor levels in rats with lower limb arteriosclerosis obliterans (ASO). Sixty rats were randomly divided into control, model, simvastatin, low-PUE, middle-PUE and high-dose PUE group. The animals in later 5 groups were with lower limb ASO, and the later 4 groups were given 1 mg/kg simvastatin and 5, 10 and 20 mg/kg PUE, respectively. The blood lipid and inflammatory factor levels were determined. Results showed that, the serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and high-sensitivity C-reactive protein (hs-CRP) level in model group were significantly increased (P 0.05). PUE can obviously decrease the blood lipid and inflammatory factor levels in rats with lower limb ASO.

Target protein candidates of hypothalamus in aging rats with intervention by Qiongyugao

To explore the associated proteins of the hypothalamus in aging rat models with intervention by Qiongyugao(QYG) based on iTRAQ technology, find out the target protein candidates and investigate the mechanism of delaying aging for Qiongyugao. The results showed that Qiongyugao increased GSH-Px activity in serum and SOD activity in liver; the total protein count identified by iTRAQ was 3 522, FDR<1%. There were 20 kinds of differential proteins between the blank group and model group; there were 295 kinds of differential proteins between model group and QYG group, and 40 kinds of them had a difference multiple ≥1.30 (the maximum value was 1.47). Compared with blank group, there were 14 kinds of proteins that were down-regulated in model group and up-regulated in QYG group. Combined with literature search and gene function search, 12 kinds of target protein candidates were screened out ： ST18, Ptprc, PSMB8, INPP4B, Shc3, Pik3r1, PIP5K1C, Nampt, Rasgrp2, Asah2, Pdpk1, and Map2k7. The expression of nuclear factor-kappa B (NF-κB) in the hypothalamic inflammatory pathway was detected by Western blot and the results showed that its expression level in model group(0.96) was higher than that in control group(0.85), while its expression level in QYG group(0.89) was lower than that in model group. Q-PCR results showed that the relative mRNA expression levels of PIP5K1C and Ptprc in model group were significantly lower than those in blank group(P<0.01); while compared with the model group, the mRNA expression levels of PIP5K1C and Ptprc in QYG group were significantly increased(P<0.01) . This result was consistent with proteomics data. QYG may delay aging by regulating hypothalamic inflammatory reaction.

The effect of picroside II on the ERK1/2 signal transduction pathway and its neuroprotective effect on the cerebral ischemic injury in rats

Objective To explore the neuroprotective effect and mechanism of picroside II on ERK1/2 signal transduction pathway after cerebral ischemia injury in rats. Methods The focal cerebral ischemic models were established by inserting a monofilament threads into middle cerebral artery occlusion (MCAO) in 100 Wistar rats and treated by injecting picroside II (20 mg/kg) intraperitoneally. The neurobehavioral function was evaluated by modified neurological severity score points (mNSS) test. The cerebral infarct volume was measured by tetrazolium chloride (TTC) staining. The apoptotic cells were counted by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The expression of pERK1/2 in cortex was determined by the immunohistochemistry (IHC) and Western Blot (WB). Results mNSS test showed that severe neurological dysfunction was found in model and LPS groups, and the scores of mNSS were significantly increased; meanwhile the scores of mNSS in treatment group and U0126 group were significantly lower than that in model and LPS groups (P<0.05). TUNEL assay showed that the apoptotic cell indexes (ACI) in different groups were (0.06±0.02), (0.27±0.03), (0.07±0.02), (0.26±0.03)and(0.09±0.05), and the ACI in treatment and U0126 groups was obviously lower than that in model and LPS groups (P<0.05). With IHC and WB, pERK1/2 level in model group was the highest, which was slightly higher than that of LPS group, and pERK1/2 expression in treatment and U0126 groups was significantly decreased compared with that in model and LPS groups (P<0.05). Conclusion The activation of ERK1/2 by cerebral ischemia could induce the cell apoptosis. Picroside II might reduce cell apoptosis by inhibiting the activation of ERK1/2 in ischemic brain injury. Key words: Picroside II; Cerebral ischemia; Apoptosis; ERK1/2; Rats

Protective effect of fasudil hydrochloride against acute renal injury in septicopyemia rats.

To observe the protective effect of fasudil hydrochloride against acute renal injury in septicopyemia rats. A total of 60 Wister rats were included in the study and divided into control group (n=10), model group (n=25) and treatment group (n=25). Model group and treatment group received intraperitoneal injection of endotoxin (ET) to establish acute renal injury models while the control group only received daily intraperitoneal injection of normal saline 1mL. Five rats were taken out of model group and treatment group respectively at 1h (T1), 6h (T2), 12h (T3), 24h (T4) and 48h (T5), for intraperitoneal injection of ET 30mg/kg. Treatment group received intraperitoneal injection of fasudil hydrochloride 30mg/kg 1h before injection of ET. For three groups, 5mL blood samples were collected from postcava for determination of serum creatinine and urea nitrogen levels at different time points. Concentrations of serum tumor necrosis factor α and ET-1 were determined by using ELISA. The renal pathologic changes were observed under the microscope. Serum creatinine levels in both model group and treatment group were significantly higher than control group at T2-T5 (P<0.05) while the levels in treatment group were significantly lower than control group at T3-T5 (P<0.05). At T2-T5, blood urea nitrogen levels in model group and treatment group were significantly higher than control group (P<0.05) while the levels in treatment group were significantly lower than model group at T3-T5 (P<0.05). Concentrations of serum tumor necrosis factor α in model group and treatment group were significantly higher than control group at T1-T5 (P<0.05) while the levels in treatment group were significantly lower than model group at T1-T5 (P<0.05). Serum ET-1 concentrations in model group and treatment group were significantly higher than control group at T1-T5 (P<0.05) while the levels in treatment group at T1-T4 were significantly lower than model group (P<0.05). Rats in control group showed no swelling or hyperemia in kidney cells but normal structure and normally arranged renal tubular epithelial cells. Obvious injury was observed in model group at T3 and renal tubular epithelial cells in disorder and at swelling condition, hyperemia and angiectasis in glomerulus, degenerative opacities and vacuolar degeneration, and maximized injury were observed at T4. Injury in renal tissue in treatment group was significantly milder than model group. Fasudil hydrochloride has the significantly protective effect against acute renal injury in septicopyemia rats.

Effect of Bizhongxiao decoction and its dismantled formulae on IL-1 and TNF levels in collagen-induced arthritis in rat synovial joints

BackgroundRheumatoid arthritis (RA), a chronic autoimmune disease, affects sufferers in many different ways. Treatment of this chronic condition is particularly challenging. Traditional Chinese Medicine (TCM) provides alternatives. Bizhongxiao decoction (BZX) is a TCM complex, which has been used clinically for many years to treat RA. The purpose of this study is to compare the effects of BZX decoction and its dismantled formulae on IL-1 and TNF-1 levels in rats with RA, and to elucidate its mechanism of action.MethodsNinety healthy normal female SD rats were randomly divided into six groups: normal (control), model, BZX decoction, and the three dismantled formulae (I: heat-clearing and detoxication, II: dissipating dampness, and III: blood circulation promotion). Apart from the normal (control) group, the rats in each group were injected subcutaneously with bovine type II collagen and complete Freund adjuvant to establish a collagen-induced arthritis model, so that inhibition of foot swelling in the rats by BZX decoction and its dismantled formulae could be observed. Immunohistochemistry was used to assess the levels of the inflammatory cytokines IL-1 and TNF in synovial joints at various time points.ResultsTwenty-one days after the model was established, the levels of TNF and IL-1 were significantly higher in the model group, BZX decoction group and dismantled formula groups I, II and III than in the normal controls (P < 0.05). The levels of these cytokines were significantly higher in the model group than the BZX decoction or the three dismantled formula groups (P <0.01). At longer times, the TNF and IL-1 levels in model group rose gradually; those in the BZX decoction and dismantled formula groups were gradually reduced. The cytokine levels in the BZX decoction group were lower than in the three dismantled formula groups and continued to decline.ConclusionsBZX decoction and the three dismantled formulae examined down-regulated the inflammatory factors IL-1 and TNF in collagen-induced arthritis rat models, but BZX exerted the strongest effect.

Increased intestinal permeability in pathogenesis and progress of nonalcoholic steatohepatitis in rats

To investigate whether increased intestinal permeability contributes to the pathogenesis and progress of nonalcoholic steatohepatitis by observing its dynamic change in rat models. Rat models of nonalcoholic steatohepatitis were established by giving a fat-rich diet. The rats were sacrificed at wk 8, 12 and 16 during the study. Rats fed with normal diet were taken as control. Plasma D-lactate, plasma diamine oxidase, serum lipids and liver transaminases were measured in blood of the femoral artery. Hepatic steatosis and inflammation were assessed by haematoxylin-eosin staining. A rat model of nonalcoholic steatohepatitis was established successfully. Plasma D-lactate level in model group at wk 8, 12 and 16 and diamine oxidase level in model group at wk 12, 16 increased significantly compared with those in control group. There were notable differences of D-lactate and diamine oxidase level in model group between wk 8 and 12 as well as between wk 12 and 16. Serum lipids, liver transaminases and liver injury also increased with disease development. Increased intestinal permeability caused by intestinal bacterial overgrowth and endotoxin-induced intestinal destruction exists in rats with nonalcoholic steatohepatitis, which may partially explain the pathogenesis and progress of this disease.