Silk fibroin/collagen composite hydrogels with different matrix stiffness influence the growth and phenotype of human mammary epithelial cells / 生理学报

Extracellular matrix (ECM) stiffness is closely related to the physiological and pathological states of breast tissue. The current study was aimed to investigate the effect of silk fibroin/collagen composite hydrogels with adjustable matrix stiffness on the growth and phenotype of normal breast epithelial cells. In this study, the enzymatic reaction of horseradish peroxidase (HRP) with hydrogen peroxide (H2O2) was used to change the degree of cross-linking of the silk fibroin solution. The rotational rheometer was used to characterize the composite hydrogel's biomechanical properties. Human normal mammary epithelial cell line MCF-10A were inoculated into composite hydrogels with various stiffness (19.10-4 932.36 Pa) to construct a three dimensional (3D) culture system of mammary epithelial cells. The CCK-8 assay was applied to detect the cell proliferation rate and active states in each group. Hematoxylin-Eosin (HE) staining and whole-mount magenta staining were used for histological evaluation of cell morphology and distribution. The results showed that with the increase of matrix stiffness, MCF-10A cells exhibited inhibited proliferation rate, decreased formation of acinus structures and increased branching structures. Meanwhile, with the increase of matrix stiffness, the polarity of MCF-10A cells was impeded. And the increase of matrix stiffness up-regulated the expression levels of mmp-2, mmp-3, and mmp-9 in MCF-10A cells. Among the genes related to epithelial-mesenchymal transition (EMT), the expression level of the epithelial marker gene E-cadherin was significantly down-regulated, while the interstitial cell marker gene Vimentin was up-regulated, and the expression levels of Snail, Wnt5b and Integrin β1 in the Wnt pathway were up-regulated. These results suggest that the silk fibroin/collagen composite hydrogels with adjustable matrix stiffness regulates the proliferation and the phenotype of MCF-10A cells. The effects of increased matrix stiffness may be closely related to the changes of the polar structures and function of MCF-10A cells, as well as the occurrence of ECM-remodeling and EMT.

Effect of Fluid Sheer Stress on Atherosclerosis and Progress of Research on Treatment of Sheer Stress Injury with Traditional Chinese Medicine： A Review / 中国实验方剂学杂志

Cells in the arterial wall are constantly subjected to the shear stress generated by the blood flow. Shear stress plays a pivotal role in the formation of atherosclerosis. The endothelial cells located between the blood and the vessel wall have a unique response to the shear stress of the blood flow， which can convert mechanical stimulation into intracellular signals， thereby affecting the pathological process of atherosclerosis. Endothelial function is not only regulated by hormones， growth factors and other biochemical substances， but also affected by mechanical forces such as blood flow shear stress. Physiologically， shear stress can play an anti-atherosclerotic role in maintaining the homeostasis of endothelial cells. Pathological shear stress will lead to endothelial dysfunction and promote the progression of atherosclerosis. Under the mediation of different shear stress， the endothelial function can be regulated through epigenetic pathways or mechanically sensitive cation channels. Therefore， it is necessary to understand how various signal transduction pathways are affected by pathological shear stress， so as to cause endothelial dysfunction and atherosclerosis. Traditional Chinese medicine（TCM） has been increasingly recognized for its curative effect in treating atherosclerosis， with the advantages of few side effects， multiple targets and multiple mechanisms. In recent years， the understanding of the anti-atherosclerosis mechanism of TCM mediated by shear stress has gradually deepened. This review will take endothelial function as the breakthrough point， systematically sort out the influence of shear stress on the pathological process of atherosclerosis and the related molecular mechanisms. Meanwhile， it is the first time to summarize the latest research progress of Chinese medicine against shear stress damage by sorting out the existing literature. This article mainly clarify the relationship between shear stress， endothelial function， atherosclerosis and TCM， in order to provide a theoretical basis for the clinical treatment and pathological mechanism of atherosclerosis.

Non-targeted metabolomics study and biomarker screening of prehypertensive liver-fire hyperactivity syndrome based on UPLC-Q-Exactive MS technology / 中国中药杂志

In this study, patients with prehypertensive liver-fire hyperactivity syndrome(LFHS) were selected as the research objects. The plasma samples of healthy volunteers and patients with prehypertensive LFHS were analyzed by non-targeted metabolomics based on UPLC-Q-Exactive MS. The differential biomarkers and metabolic pathways were screened out by multivariate statistics and metabolic pathway analysis, which revealed the characteristics of metabolic patterns of the syndrome. Thirty-three potential biomarkers such as androsterone and lysophosphatidylcholine and 16 related metabolic pathways such as steroid hormone metabolism and lipid metabolism were identified, and a partial least squares-discriminant analysis(PLS-DA) model of traditional Chinese medicine(TCM) syndromes was preliminarily constructed: Y =-0.070X_(13)-0.006X_8+ 0.040X_5-0.152X_1+0.131X_(10)+0.036X_(11)+0.043X_(23)+0.076X_(16)+0.132X_(20)+0.081X_(19)-0.101X_(31)+0.082X_(15)-0.038X_9+0.079X_(24). The predictive value of the model was 88.1%, and the explanatory power was 88.4%. In this study, the characteristic metabolic pattern of the prehypertensive LFHS was distinguished and revealed by metabolomics. The constructed PLS-DA model is expected to provide an objective basis for the identification of TCM syndromes in prehypertension, and inspiration for exploring the biological basis of TCM syndromes at small-molecular and overall levels.

Study on the Intervention Effects of Pinggan Prescription () on Spontaneously Hypertensive Rats Based on Metabonomic and Pharmacodynamic Methods / 中国结合医学杂志

OBJECTIVE@#To investigate the effects of Pinggan Prescription (, PGP) on hypertension by the associated methods of metabonomic and pharmacodynamic.@*METHODS@#A total of 32 male spontaneously hypertensive rats (SHRs) were randomly divided into two groups by using the random number table method: a treatment group (n=18) and a model group (n=14). The Wistar rats (n=14) were used as the normal group. Different prescription were used to intervene three groups: the treatment group in which PGP extract was administered orally at a dose of 18.336 g/kg (PGP/body weight), and the model group in which physiological saline was administered at the equivalent dose. The same treatment was applied to the normal group as the model group. The blood pressure was measured by tail-cuff method, and pharmacodynamic indexes including cyclic adenosine monophosphate (cAMP) and angiotensin II (Ang II) were tested by enzyme-linked immunosorbent assay. The plasma samples from three groups were detected by gas chromatography-mass spectrometry (GC-MS).@*RESULTS@#Compared with the model group, blood pressure of treatment group was obviously reduced after continuous curing with PGP (P<0.01). The pharmacodynamic results illustrated that the content of Ang II increased with the raised blood pressure and the cAMP expressed the converse trend. After curing with PGP, the content of Ang II decreased, the difference between model group and treatment group was significant (P<0.01), and the cAMP expressed the converse trend. Five potential biomarkers were identified, including arachidonic acid, hexadecanoic acid, elaidic acid, octadecanedioic acid and 9,12-octadecadienoic acid. These metabolites had shown significantly changes as followed: arachidonic acid, hexadecanoic acid and elaidic acid were significantly higher and octadecanedioic acid and 9,12-octadecadienoic acid were lowered in the model group than those in the normal group. After the treatment of PGP, the metabolites had the trends of returning to normal along with the reduced blood pressure.@*CONCLUSIONS@#PGP intervention for hypertension played a major role in the metabolism of arachidonic acid and linoleic acid. Metabonomic with pharmacodynamic methods could be potentially powerful tools to investigate the mechanism of Chinese medicine.

The mechanism of action of valsartan studied by HPLC-TOF/MS / 药学学报

High performance liquid chromatography-time-off-flight mass spectrometer (HPLC-TOFMS) technology coupled with partial least squares discriminant analysis (PLS-DA) processed by SIMCA-P software was applied to investigate serum endogenous metabolites alternations of valsartan in spontaneous hypertension rats (SHR). And MetPA platform was used to connect identified potential biomarkers in corresponding metabolic pathways to find possible therapeutic mechanism of valsartan. Valsartan significantly declined the blood pressure of SHRs (P < 0.05) at fourth week. The metabolic profiling significantly changed and four metabolites involved in G protein-coupled pathway were identified. Metabolomics is able to detect holistic and microcosmic alternations in organism, so as to elucidate therapeutic mechanism of drugs.

Intervention effects of qingre jiangya capsule on brain hippocampus of spontaneously hypertensive rats based on metabonomic research / 中国中药杂志

Thirty SHRs were obtained randomly to hypertension, model group, captopril group and Qingre jiangya capsule group. Ten Wistar rats were used as control group. The hippocampus tissue was removed to explore the damage of spontaneously hypertensive rats (SHR) and the protective effect of Qingre jiangya capsule after continuously administered for 14 days. And then the data were processed by principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). The research results revealed captopril group was significantly different from the other three groups. The classification of other three groups is also very clear after captopril group removed. This suggested that Qingre jiangya capsule could improve the overall metabolism compared with captopril. Four metabolites were identified: dimethylglycine, glycerophosphocholine, aldosterone and noradrenaline. Hypertension hippocampus damage may mainly be expressed in tyrosine metabolism, aldosterone-regulated sodium, vascular smooth muscle contraction reabsorption, and Qingre jiangya capsule could reverse the hippocampus tissue damage of SHR.

Improvement of naringein on cognition of model rats with Alzheimer's diseases and its mechanism / 中草药

Objective: To investigate the effect of naringein on the cognition of rats with Alzheimer's diseases (AD), its regulation on amyloid β-protein (Aβ) and Tau antibody, and its potential mechanism of improving the cognition of rats with AD. Methods: AD model of rats was established by intracerebroventricular (icv) Streptozotocin (STZ) on days 1 and 3, and then naringein and Rosiglitazone were ig administered. Rats in model and control groups were treated with the same volume of physiologic saline to induce dementia model. On day 21 after the first operation, spatial learning and memory of rats were tested in Morris water maze. The expression of insulin-degrading enzyme (IDE), glycogen synthase kinese-3β (GSK-3β), phospho-GSK-3β (pGSK-3β), Tau, and phospho-Tau (pTau) were measured by Western blotting. Aβ42 and Aβ40 levels in the brain of the AD rats were tested by immunohistochemistry. Results: Cognition of AD rats administered with naringein and Rosiglitazone could be recovered by Morris water maze (P < 0.05). Western blotting results showed that both naringein and Rosiglitazone could improve the expression of IDE and decrease the activity of GSK-3β, and reduce the phosphorylation level of Tau (P < 0.05). Immunohistochemistry demonstrated that the Aβ42 and Aβ40 in cerebral cortex of rats treated by naringein and Rosiglitazone were both decreased. Conclusion: Naringein as a peroxisome proliferator-activated receptor γ (PPARγ) excitomotor could act as insulin sensitizer, and could alleviate the cognition of AD rats, Aβ deposition, and phosphorylation of Tau through regulating the expression of insuline-related IDE and GSK-3β.