Objective To establish a cell model of insulin resistance in BRL-3A cells,and to elucidate the underlying mechanisms of palmitic acid (PA). Methods Using CCK8 method to observe the effect of different concentration and time of PA on proliferation in BRL-3A cells.The glucose consumption were detected at different concentration and time points by glucose detection kit.The expression of insulin receptor substance 1(IRS-1),phosphoinositide-3-kinase(PI3K),phosphorylated serine-threonine kinase(p-AKT),serine-threonine kinase(AKT),glucose transporter 4(GLUT4)in BRL-3A cells were detected by Western blotting. Results The 0.2 mmol·L-1 PA had no effect on proliferation in BRL-3A cells within 36 h.The insulin resistance model of BRL-3A cells can be successful established by incubating in cultured medium with PA in 3 conditions:0.4 mmol·L-1 PA for 6 h,0.25 mmol·L-1 PA for 12 h,0.2 mmol·L-1 PA for 12 h.And insulin stimulated glucose transport was inhibited by high concentration of PA in BRL-3A cells.Palmitic acid decreased IRS-1,PI3K,p-AKT,GLUT4 expression in BRL-3A cells. Conclusion High concentration of palmitic acid may induce insulin resistance in BRL-3A cells via inhibiting the expression of IRS-1 and other target proteins.
Nowadays,neither marketed drugs nor commonly used clinical therapies have been proved by evidence-based medicine in promoting neural repairment and neurogenesis after stroke. On the one hand,hippocampal injury by cerebral ischemia can lead to cognitive impairment in learning and memory. On the other hand,the subgranular zone of dentate gyrus in the hippocampus is one of the main neurogenic niches. This paper not only expounds the mechanism and process of hippocampal neurogenesis,but also tells how neurogenesis has been affected by the microenvironment of neural stem cells in the subgranular zone of dentate gyrus in the hippocampus. It also primarily summarizes the influencing effects of the active ingredients and compounds of traditional Chinese medicine on hippocampal endogenous neurogenesis and repairment after cerebral ischemia. All of these are expected to provide references for relevant research of utilizing traditional Chinese medicine to promote hippocampal endogenous neurogenesis and repair after cerebral ischemia.
Celastrol, extracted from the root of Tripterygium wilfordii,has been recognized as the most potential anti-obesity active component for its unique effects on the regulation of body weight and lipid metabolism. Based on the latest studies on celastrol all over the world,this paper reviews the role of celastrol in the regulation of lipid metabolism and provides a summary of the molecular mechanisms,including increasing the sensitivity to leptin receptors,inhibiting insulin resistance,increasing energy consumption in mitochondria,suppressing the differentiation of adipocytes,improving intestinal microecology, and anti-inflammatory effects; aiming to provide scientific references to the research and development of metabolic drugs based on celastrol.
Objective To investigate the effects of puerarin on behavior and memory-related proteins in rats with chronic stress induced by noise. Methods Thirty-two SD male rats were divided into normal control group,model control group,low dose (100 mg·kg-1)puerarin group and high dose (200 mg·kg-1)puerarin group(n=8 in each group).The rat model of chronic stress was established by continuous noise stimulation for 60 days.Sugar consumption test and water maze test were carried out.The expression of memory-related proteins,such as transient receptor potential vanilloid subfamily member 1 (TRPV1),protein kinase C (PKC),phosphorylated cAMP-response element binding protein (p-CREB)and brain derived neurotrophic factor (BDNF),in hippocampus were detected by Western blotting and ELISA. Results Compared with normal control group,the body mass of rats in the model control group and the puerarin group increased slower,and the consumption of sugar water decreased.In the water maze test,the escape latency of model control group was significantly longer than that of the normal control group, and the total swimming distance in the target quadrant was significantly decreased.The expression of TRPV1 and PKC was significantly up-regulated,and the expression of p-CREB and BDNF was significantly down-regulated.There was statistical significance (P<0.05).After treatment with 100 mg·kg-1 puerarin,the escape latency was significantly shortened,the total swimming distance in the target quadrant was increased,the expression of TRPV1 and PKC was significantly down-regulated,and the expression of p-CREB and BDNF was significantly up-regulated in the low dose puerarin group.There was statistical significance (P<0.05). Conclusion Long-term noise stress affects the learning and memory ability of the growing rats,whereas puerarin can partially improve the learning and memory ability of chronic stress rats,and its mechanism may be related to adjust the expression level of memory-related proteins in hippocampus.
Objective To investigate the inhibitory effect and its mechanism of salidroside on inflammatory response and neuronal apoptosis in depressed rats induced by chronic unpredictable mild stress (CUMS). Methods A depressed rat model was constructed by using CUMS.From week 6 to week 12,rats in the low,medium,and high dose of salidroside groups were intraperitoneally injected with 12.5,25,50 mg·kg-1 salidroside once a day;rats in the fluoxetine group were injected with 5 mg·kg-1 fluoxetine;rats in the normal control group and the model control group were injected with the same amount of 0.9% sodium chloride solution;and rats in the H-89 group were intrathecally injected with 2 mg·kg-1 H-89.Rat body mass and sucrose preference were recorded. Open field experiments,forced swimming tests,and Morris water maze were performed at 12 weeks.Neurological injury in the hippocampus was observed by hematoxylin-eosin (HE)staining.Neuronal apoptosis was detected by flow cytometry.The levels of cyclic adenosine monophosphate (cAMP),interleukin-6 (IL-6),interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were detected by enzyme linked immunosorbent assay (ELISA).The expression levels of cAMP-dependent protein kinase catalytic subunit-Ⅱα(PKAc-Ⅱα)and phosphorylated cAMP-response element binding protein (p-CREB)/CREB proteins were detected by Western blotting. Results After administration of salidroside,the weight of CUMS rats,the preference rate of syrup and the distance traveled in the open field experiment,the number of movement trajectories,and the number of standings increased significantly.The time required to force the swimming time and the time to find the platform is significantly reduced.The number of found platforms within 60 s has increased significantly.The neuronal apoptosis rate and the levels of IL-1β,IL-6 and TNF-α in hippocampus were significantly reduced.The content of cAMP in hippocampus increased significantly.The expression of PKAc-Ⅱα/β-actin and p-CREB/CREB protein was significantly up-regulated.Addition of PKA inhibitor H-89 can significantly reverse the above changes in CUMS rats caused by salidroside. Conclusion Salidroside may inhibit inflammatory response and neuronal apoptosis in CUMS-induced depression rats through cAMP/PKA/CREB signaling pathway.
Objective To investigate the membrane permeability of pulsatillae pentacyclic triterpenoid saponins (pulchinenosides) B3,BD,B7,B10,B11 in duodenum,jejunum and ileum and the effect of organic anion transport polypeptides (OATPs) on the absorption of pulchinenosides in the intestinal epithelial cells. Methods Rat in-situ single-pass intestine perfusion model and ultra-high performance liquid chromatography-triple quadrupole-linear ion trap-mass spectrometry (UPLC/Q-TRAP-MS) were adopted via the co-administration of OATPs inhibitors findomethacin and naringenin. Results The permeability of pulsatilla saponins in the duodenum was significantly greater than that of the jejunum and ileum (P<0.05,P<0.01) and indometacin and naringenin inhibited the absorption of pulchinenosides in duodenum,jejunum and ileum (P<0.05,P<0.01). Conclusion The duodenum was probably the main site for the absorption of pulchinenosides and pulchinenosides was probably the substrates of the OATPs.
Objective To investigate the effects of sinomenine (SIN)on lipopolysaccharide (LPS)induced injuries in rat H9c2 cardiomyocyte and its possible mechanism. Methods The cells were randomly divided into normal control group,LPS group and sinomenine intervention group(25,50 and 100 μmol·L-1).After treatments,the mRNA levels of pro-inflammatory factors in each group were detected by quantitative real-time PCR.Meanwhile,to detect the effect of sinomenine on apoptosis,we observed the levels of apoptosis-related proteins (Bax,Bcl-2,caspase 3,and caspase 9),ERK and MEK by Western blotting.In addition,we utilized Western blotting to examine the levels of MEK and ERK as well. Results Compared with LPS group,the mRNA levels of TNF-α,IL-6,IL-1β and MCP-1 were significantly increased (P<0.05),which were apparently inhibited by sinomenine (P<0.05).And sinomenine observably ameliorated apoptosis,which was fostered by the lower levels of apoptosis-related proteins compared to LPS group (P<0.05).In addition,sinomenine reversed the activation of ERK and MEK as well (P<0.05). Conclusion Sinomenine may constitute a protective role in cardiomyocyte following LPS through inhibiting inflammatory response and apoptosis.
Objective To explore the effect and mechanism of ursolic acid (UA) extracted from Hippophae rhamnoides L.on alcohol-induced liver disease in rats. Methods Fifty male Wistar rats (SPF)were randomly divided into five groups:normal control group,model control group,low-,medium- and high-dose UA group,10 rats in each group.The rats were administered by intragastric administration for eight consecutive weeks.After 12 hours of last administration,blood was collected by abdominal aorta and the liver was obtained for histopathological and biochemical assays.The levels of ALT and AST in serum was detected.The content of TG in liver homogenate was determined.The morphological changes of hepatic tissue in rats were observed by HE staining.The apoptotic rate of rat liver cells was detected by TUNEL method.The protein expressions of Bcl-2,Bax and cleaved caspase-3 in liver tissue were detected by Western blotting. Results In model control group,HE staining showed that the hepatic cord was arranged disorderly,and there were lots of vacuoles in the cytoplasm of hepatocytes.The inflammatory cells in the liver tissue gathered,and the hepatocytes showed flake necrosis in model control group. Compared with model control group,ursolic acid significantly improved the steatosis of the liver.In the UA group,infiltration and necrosis of inflammatory cells were significantly decreased,the tissue structure tended to be normal,and the activity of serum ALT,AST,and the content of TG in the medium- and high-dose UA group liver tissue were significantly decreased (P<0.05).TUNEL results showed that the number of positive cells and the apoptosis index of hepatocytes in the high dose of UA group were significantly lower than in model control group (P<0.05).Western blotting results showed that the protein expression of Bcl-2 in liver tissue of UA groups was significantly increased,the protein expression of Bax was significantly decreased,and the protein expression of cleaved caspase-3 was increased,compared with model control group. Conclusion Ursolic acid extracted from Hippophae rhamnoides L.could improve the liver injury induced by alcohol.The mechanism might be related to the regulation the protein expressions of Bcl-2,Bax and cleaved caspase-3,key proteins of apoptosis,thereby inhibiting the abnormal apoptosis of hepatocytes.
Objective To predict the target of the main active ingredients of Honghua sanwei san and to explore the therapeutic mechanism of non-alcoholic fatty liver disease (NAFLD)through its multi-component,multi-target,and multi-pathway. Methods The active ingredients of Honghua sanwei san were collected by traditional Chinese medicine systems pharmacology (TCMSP)database and screened by oral bioavailability (OB)and pharmacodynamics (DL).The potential targets of compounds were predicted by reverse pharmacophore matching method and drug similarity method,and then the compound-target network was constructed.NAFLD disease-related targets were collected by online Mendelian inheritance in man (OMIM)database and DisGeNET database,and then disease target interaction network diagram was constructed.Finally,the biological function and related signal pathway were analyzed by DAVID online tool. Results Network analysis results showed that 47 active compounds screened from Honghua sanwei san directly or indirectly acted on 42 key targets of FAFLD disease.A total of 17 biological processes and 13 KEGG pathways were involved (P<0.05,false discovery rate<0.05),suggesting that Honghua sanwei san was mainly used to treat NAFLD through the intervention of NAFLD,insulin resistance,hepatitis B,PPAR signaling pathway,TNF signaling pathway and MAPK signaling pathway. Conclusion The material basis and the core target of Honghua sanwei san were preliminarily predicted by network pharmacology,which provided scientific basis for further elaborating the mechanism of Honghua sanwei san in treating NAFLD.
Objective To study the clinical efficacy and safety of combination therapy of intravitreal aflibercept and sub-threshold micropulse laser (SMPL)for diabetic macular edema (DME). Methods This was a single-center prospective randomized controlled trial recruited 60 patients(70 eyes)with DME.These eyes were randomized into 2 different groups. Thirty-five eyes of 30 patients who received aflibercept injections alone (IVA)were included in group A.Thirty-five eyes of 30 patients in group B received combined aflibercept injections followed by SMPL(IVA+SMPL).The primary outcomes was the mean change in best-corrected visual acuity (BCVA)and central macular thickness (CMT)in the followed 12 months.Secondary outcomes include the number of aflibercept injections and any recorded complications. Results At 12 months,31 eyes in the group A and 33 eyes in the group B fulfilled the treatment.Final BCVA in both groups showed statistically significant improvement (P<0.05)comparing to the baseline visual acuity,but without statistically significant difference between the two groups (P>0.05).There was significant reduction in CMT in both groups (P<0.05). The CMT were significantly lower in group B than group A (P<0.05).The number of injections were significantly lower in group B (4.9±1.6)than group A (7.3±1.5)(P<0.05).In both groups,no complications related to the micropulse laser and aflibercept injections were encountered. Conclusion The combination therapy of aflibercept and SMPL is a safe and effective treatment for DME,by decreasing the frequency of aflibercept injections while maintaining good visual acuity.
Tripterygium wilfordii is a common traditional Chinese medicine used in the treatment of autoimmune diseases,which has serious toxicity and side effects,and even causes death.In the article,we summarized the toxic components,molecular mechanisms,clinical manifestations and strategy for reducing toxicity of Tripterygium wilfordii, from the records in historical and modern literatures,which might provide guidance for the rational use of Tripterygium wilfordii.
Coronary no-reflow is a common complication of reperfusion therapy.The prognosis of patients will be seriously affected.Many studies have proved that statins can not only reduce blood lipid,but also improve myocardial blood perfusion after percutaneous coronary intervention (PCI),prevent and reduce no-reflow phenomenon,and improve cardiac function.The mechanism of action mainly includes anti-inflammation,improving endothelial function,up-regulating endothelial nitric oxide synthase level,promoting nitric oxide synthesis and anti-apoptosis.
The treatment of non-alcoholic steatohepatitis (NASH)is extremely lacking,there is only one approved drug in the world,Saroglitazar.The Farnesoid X receptor (FXR)agonist can effectively reduce the accumulation of lipids and the inflammation in the liver,which has the potential to be a drug for the treatment of NASH.At present,there are only 6 FXR agonists for NASH indications (obeticholic acid,EDP-305,cilofexor,tropifexor,LMB763,PXL007)that have entered the clinical research stage globally.This article reviewed the pharmacological research of these 6 compounds to provide a basis for the development of NASH drugs targeting FXR.
Objective To establish a combination approach of magnetic solid-phase extraction,the transcriptomic preparation and liquid chromatography coupled with triple quadrupole and time-of-flight mass spectrometry (triple TOF LC-MS/MS)technology,and to identify anti-thrombosis biomolecules from H.nipponica. Methods Thrombin-immobilized nanobeads were synthesized and employed to screen anticoagulants from leech saliva.A customized transcriptomic database was constructed to search against the peptide MS/MS spectrum for antithrombin biomolecule identifications. Results The Fourier transform infrared (FT-IR)spectrum and transmission electron microscope (TEM)image indicated that all modifications were successfully immobilized on the surface of materials.Within 19 days at 37 ℃,the storage stability of immobilized thrombin was satisfactory with the relative standard deviation 4.74%.Five direct thrombin inhibitors,all referring to hirudin variants,were successfully enriched and identified.The antithrombin activity was 70 U·g-1. Conclusion By incorporating magnetic solid-phase extraction,the transcriptomic preparation with triple TOF LC-MS/MS technology,a convenient and easily-handled method is successfully developed to screen and identify the antithrombin biomolecules from H.nipponica.This efficacious application also show its practical and bright future in biomolecules discovery from animal medicines.
Objective To find out a mildew marker of Helianthi annui receptaculum and to assess its application study in safety evaluation of traditional Chinese medicine. Methods The mildew marker of Helianthi annui receptaculum was isolated and prepared by column chromatography and thin layer chromatography (TLC) . An evaluation method for Helianthi annui receptaculum was established based on TLC technology. Twelve kinds of mildew Chinese medicine were collected to evalute the applicability of the method. Results TLC of mildew samples, including Helianthi annui receptaculum,Flos Sophora,Pericapium Citri Reticulate and Panax ginseng showed the same fluorescence compared with mildew markers. Conclusion The established method for evaluating the coiledew of sunflower is sensitive and specific.It has certain applicability to the evaluation of mildew of plant medicinal materials with high content of polysaccharides,saponins and proteins,but it is not suitable for medicinal fungi.
Objective To evaluate the germicidal effectiveness and resistance risk for 15 kinds of meropenem dosage regimens. Methods We used PK/PD model,Monte Carlo simulation,and MIC distribution in intensive care unit (ICU)and non-ICU,to evaluate the probability of target attainment and cumulative fraction of response (CFR)of 15 kinds of dosage regimens for Escherichia coli(EC),Klebsiella pneumonia (KP),Pseudomonas aeruginosa (PA)and Acinetobacter baumannii (AB)in effectiveness and the ability to suppress resistance mutation.(%T>MIC>40% in non-ICU,%T>MIC>100%,%T>4MIC>50% in ICU as target threshold, %T>4MIC>100% both in non-ICU and ICU as target threshold.) Results For non-critical patients,%T >MIC >40% was taken as the target threshold of effectiveness.For EC and KP,with the exception of 0.5 g, q12 h,the CFR of all the other regimens were greater than 90%.For PA,only 2 g q6 h and 3 g q6 h could achieve 90% for CFR.For AB,only 3 g q6 h could reach 90%.For the critical patients,whether %T>MIC >100% or %T>4MIC >50% was taken as the target threshold of effectiveness,the results were similar.For EC and KP,only part of the regimen CFR could reach 90%.For PA and AB,the CFR of all regimens couldn't reach 90%,even if the 3 g q6 h regimen could only reach 62.6% for PA,and for AB could only reach 11.2%.In order to achieve the purpose of inhibiting drug-resistant mutations,a more stringent target threshold (i.e.,T>4MIC >100%)was formulated.According to the MCS results,it was difficult to reach the standard for non-critical patients and critical patients.Only the 3 g q6 h regimen in critical patients could achieve 90% CFR for target treatment against EC and KP. Conclusion The current pattern of meropenem use is an important factor contributing to the spread of meropenem resistance.It is need to further optimize the current infusion pattern in order to improve the treatment effectiveness and minimize the risk of drug resistance mutations.
Objective To evaluate the effect of pharmaceutical care provided by clinical pharmacists in schizophrenia patients treated with clozapine. Methods Clinical pharmacists provided pharmaceutical care for 3 patients with schizophrenia during their oral administration of clozapine. The adverse reactions,drug-drug interactions and drug-food interactions were monitored by methods of therapeutic drug monitoring,and gene polymorphism detection of metabolic enzymes. Results Clinical pharmacists assisted doctors in solving the difficult problems in the clinical use of clozapine,scheming the individualized drug treatment,and monitoring the patient's medication process. Pharmaceutical care helped to find,prevent and solve problems related to medication,to ensure the patient's medication safety. Conclusion Pharmaceutical care played an important role in long-term maintenance treatment of schizophrenia.
Objective To understand the inpatient clinical characteristics of drug-induced liver injury (DILI)in a hospital and to provide reference for the clinical prevention and treatment of DILI. Methods The medical records of DILI inpatients in a hospital in recent 5 years were retrospectively analyzed,including age,laboratory examination,use of hepatoprotective drugs,and other information. Results The majority of the 72 patients were female and over 40 years old with the Han nationality accounting for 88.89%.The top three drugs causing DILI were divided into 28 cases of traditional Chinese medicine (38.89%),26 cases of anti-tumor drugs (36.11%),and 3 cases of antibacterial drugs (4.17%).The main route of administration was oral administration (59.72%),and the median time from medication to onset was 10 days.The clinical manifestations were poor appetite,weakness,skin sclera yellow dye,greasy,and nausea.In liver function examination,the most obvious change was the elevation of ALT and AST.The most common DILI type was hepatocyte injury type and the highest proportion of grade 1 was hepatic injury (54.17%).Liver protecting drugs were mainly used in two or more combinations.Most patients with DILI had a better prognosis and 48.61% were cured,improved and effective. Conclusion DILI is caused by a variety of drug and its clinical manifestations is lack of specificity.More attention should be paid to DILI and its clinical characteristics to promote rational drug use in clinic.
This paper introduces the regulatory system of authorized generic drugs in the United States (US),analyzes the differences between China and the US,and discusses its possible impact on China,to promote relevant researches. Authorized generics,as a special type of generics,is an important tool for the original drug enterprises to occupy the market share in the US. For our country,it may be harmful to the generic competition,and reduce the enthusiasm of pharmaceutical enterprises. In the contrast, it can be used to solve public health emergencies at special periods,and to force the pharmaceutical enterprises to take the initiative to transform and upgrade. Sale of authorized generics is a common strategy in the mainstream market. Everything has its two sides,the pharmaceutical enterprises of our country need to improve our competitiveness,and be ready to cope with the competition.