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In recent years,assisted reproductive technology(ART)is developing rapidly,especially in vitro fertilization and embryo transfer(IVF-ET)technology.The rate of clinical pregnancy in our country is about 40％,and how to improve the rate of clinical pregnancy of IVF-ET has been the focus of the scholars research content.Environmental factors including culture system,temperature,humidity,pH value and volatile organic compounds,all of these can affect the success of IVF-ET.Now the common environmental factors affecting IVF-ET research progress were reviewed as follows.
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Objective To report a newly developed method and procedure to establish a rat model of subarachnoid hemorrhage in detail, and to provide a better model simulating the clinical subarachnoid hemorrhage caused by a ruptured aneurysm for related research.Methods One hundred and twenty healthy SPF 2-3-month old male Sprague-Dawley rats were divided into 4 groups, 30 rats in each group.The three experimental groups were sacrificed at 6, 24 and 72 hours after modeling.Rat models of subarachnoid hemorrhage were established by inserting a fiber core in the internal carotid artery and piercing this artery.Successful establishment of the subarachnoid hemorrhage model was confirmed by observation of breathing, pupil, defecation, urination and inspection at autopsy dissection.The controllability and reproducibility of this model were verified by observation of clinical manifestation and explored by mortality analysis.Results Subarachnoid hemorrhage was successfully induced by fiber core piercing the internal carotid artery at the needed location.Conclusions This method of model preparation is stable and understandable.The operation is nimble, with a good reproducibility.This model can be successfully performed after a short time learning, well simulate the sudden hemorrhage caused by a ruptured aneurysm, and suitable for research on early brain injury and vasospasm after subarachnoid hemorrhage.
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Objective To explore the effect of extracellular regulated protein kinases (ERK) signaling pathway on early brain injury and autophagy of nerve cell in hippocampus area in rats with subarachnoid hemorrhage (SAH). Methods Forty-eight adult male Sprague-Daw-ley rats were randomly divided into sham group, SAH group, SAH+dimethyl sulfoxide (DMSO) group and SAH+U0126 group, with 12 rats in each group. The SAH model was established with puncture of internal carotid artery. The SAH+U0126 group was injected with U0126 0.05 mg/kg;the sham group and SAH group were injected with normal saline, and the SAH+DMSO group was injected with DMSO 30 min-utes before modeling. They were sacrificed 24 hours after modeling. The brain water content was measured with wet and dry method. The morphology changes of neural cells in hippocampus CA1 were observed by HE staining. The expression of phosphorylation ERK (p-ERK), Beclin-1 and LC3-Ⅱwere detected with immunohistochemical method and Western blotting. Results Compared with the sham group, the brain water content increased (P0.05). Conclusion The activation of ERK signaling pathway may alleviate early brain injury after SAH by regulation of autophagy.
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<p><b>OBJECTIVE</b>To explore the inhibition effect and mechanism of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP)on myofibroblast differentiation via regulating acetylated tubulin α (Ac-Tub α)in vivo and in vitro.</p><p><b>METHODS</b>Silicotic model were made by SiO2 douched and divided into 6 groups as follows: control (4w, 8w)group, silicotic model (4w, 8w)group and post-or pre-treatment by Ac-SDKP group. Pulmonary fibroblasts were divided into 5 groups: (1) control; (2) Ang II; (3) Ang II+Ac-SDKP; (4) Ang II+Valsartan; (5) Ang II+TCS histone deacetylase (HDAC)6 20b. The localization of Ac-Tub α and α-smooth muscle actin (SMA) were observed by immunohistochemical (IHC) and immunofluorescence staining. The protein levels of Ac-Tub α, α-SMA, collagen type I (col I) and HDAC6 were measured by western blot.</p><p><b>RESULTS</b>In silicotic nodules and interstitial fibrosis area, positive expression of α-SMA, a classical marker of myofibroblast, was ob-served by IHC, accompanied with absence expression of Ac-Tub α. Furthermore, Ac-SDKP post-treatment could attenuate the levels of col I, α-SMA and HDAC6 to 48.39%, 52.63% and 70.18% compared with the silicotic 8w group respectively. And in Ac-SDKP pre-treatment group, compared with the silicotic 8w group, these protein levels were decreased to 32.26%, 64.91% and 54.39% respectively (P<0.05). The up-regulation of Ac-Tub α was found in Ac-SDKP post-and pre-treatment and increased to 3.00 and 2.90 folds compared with the silicotic 8w group. Compared with control group, the levels of α-SMA, HDAC6 and col I in Ang II group were up-regulated to 1.66, 3.56 and 4.00 folds accompanied with down-regulation of Ac-Tub by 44.44% (P<0.05). Pre-treatment with Valsartan, TCS HDAC6 20b or Ac-SDKP could inhibited all this changes induced by Ang II in vitro.</p><p><b>CONCLUSION</b>Ac-SDKP can inhibit the myofibroblast differentiation and collagen deposition via sup-press HDAC6 and up-regulate the expression of Ac-Tub α in vivo and in vitro.</p>
Subject(s)
Animals , Rats , Actins , Metabolism , Cell Differentiation , Collagen Type I , Metabolism , Disease Models, Animal , Fibroblasts , Cell Biology , Lung , Pathology , Myofibroblasts , Cell Biology , Oligopeptides , Pharmacology , Silicon Dioxide , Toxicity , Silicosis , Drug Therapy , Tubulin , MetabolismABSTRACT
[ ABSTRACT] AIM:To observe the effect of oleanolic acid ( OA) on the expression of Tumor necrosis factor-α( TNF-α) and collagen in silicotic rats in vivo and its possible mechanism.METHODS:Male Wistar rats were divided in-to 4 groups according to the randomized block design:control group, model group, OA group and solvent control group (20 rats in each group) .Except control group, the rats in other groups were induced by intratracheal instillation of silicon di-oxide (SiO2;250 mg/kg).The rats in OA group were intragastrically administered with OA (60 mg/kg) from the second day of giving SiO2 .The rats in solvent control group were gavaged daily with 0.6%sodium carboxymethyl cellulose solution (10 mL/kg).The rats in control group were given normal saline under the same condition for 56 consecutive days.All rats were killed at the 7th, 14th, 28th and 56th days.The lung coefficient was detected and the morphological changes were ob-served.The serum contents of TNF-αwere detected by ELISA.The content of total collagen in the lung tissue was meas-ured.The protein level of nuclear factor-κB ( NF-κB) in the lung tissue was determined by immunohistochemical method. RESULTS:(1) According to the morphological changes, the silicosis model was successfully established.Compared with control group, the lung coefficient and total collagen increased obviously in model group and solvent control group.The lung coefficient and total collagen content in OA group at each time point reduced compared with those in model group and sol- vent group, and increased compared with those in control group at the corresponding time points.(2) The serum contents of TNF-αin model group and solvent control group significantly increased, peaking at the 14th day, slightly decreasing af-terward, and showing statistically significant difference at each time point compared with those in control group.No signifi-cant difference between model group and solvent group at different time points was observed.OA had inhibitory effect on the contents of TNF-αcompared with model group and solvent group at the corresponding time points.(3) NF-κB in model group and solvent control group significantly increased, peaking at the 28th day, and showing statistically significant differ-ence at each time point compared with those in control group.The NF-κB expression in OA group was similar to model group, but significantly decreased compared with control group at each time point.CONCLUSION: OA inhibits the ex-pression of TNF-αand collagen and attenuates the silicosis fibrosis, which may be related to the NF-κB pathway.
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<p><b>OBJECTIVE</b>To explore the inhibition effect of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) on myofibroblast differentiation of MRC-5 human fetal lung fibroblasts induced by angiotensin (Ang) II.</p><p><b>METHODS</b>The study was divided into 2 step: (1) MRC-5 human fetal lung fibroblasts was induced for 48 h at different dose of Ang II and at different time point by 100 nmol/L Ang II. Then the expression of collagen type I and α-smooth muscle actin (α-SMA) were mesaured by western blot. (2) MRC-5 human fetal lung fibroblasts were divided into 4 group: (1) control, (2) Ang II, (3) Ang II+Ac-SDKP, (4) Ang II+8-Me-cAMP (a specific activator of Epac). The α-SMA expression was observed by immnocytochemical stain. The protein expression of collagen type I, α-SMA, serum response factor (SRF), myocardin-related transcription factor (MRTF)-A, exchange protein directly activated by cAMP (Epac) 1, 2 were measured by Westen blot.</p><p><b>RESULTS</b>Myofibroblast differentiation could be induced by Ang II from MRC-5 cells with a dose- and time-dependent manner. The up-regulation of SRF and MRTF-A were observed in MRC-5 cells induced by Ang II and accompanied with collagen I and α-SMA increased. Pre-treatment with 8-Me-cAMP or Ac-SDKP could attenuated all this changes induced by Ang II, and promoted the expression of Epac1.</p><p><b>CONCLUSION</b>Ac-SDKP can inhibit the myofibroblast differentiation of MRC-5 cells induced by Ang II via Epac1 activating.</p>