Effects of Diazepam,Phenobarbital,Propranolol,and Cimetidine on Diazepam Oxidizing Isoenzymes in Rat Liver Microsomes

Isolation and identification of the liver microsomal cytochrome P 450 isoen zymes responsible for the formation of diazepam main metabolites nordiazepam and temazepam in rats were studied. The effects of P 450 inducers and inhibitors on the protein contents in SDS poly acrylamide gel electrophoresis and thin layer chromatography to the corresponding diazepam me tabolizing activities of rat liver microsomes were observed. The P 450 contents were dramatically re duced by ip diazepam, cimetidine or propranolol. Diazepam and propranolol inhibited temazepam formation, high dose of propranolol also inhibited nordiazepam formation. Phenobarbital increased the P 450 contents and induced the production of both nordiazepam and temazepam. It also induced proteins with molecular weight (m) of 51 and 59 kDa in SDS PAGE and those with m ranging from 45 to 55 kDa and from 55 to 65 kDa in TLC. Propranolol inhibited both fractions, especially that of m 55～65 kDa, whereas diazepam tended to inhibit the fraction of 45～55 kDa. The protein of m 51 kDa could be mainly involved in diazepam C3 hydroxylation, whereas those of m 59 kDa could be responsible for the N demethylation of diazepam in rats.

Effect of single-use versus combined-use moschus and diazepam on expression of amino acid neurotransmitters in the rat corpus striatum

The present study analyzed expressional changes of excitatory neurotransmitters and inhibitory neurotransmitters in the rat corpus striatum after single-use and combined-use diazepam and Chinese herb moschus. The influence of moschus on the central nervous system was analyzed, in particular whether moschus increased penetration of other drugs into the brain. Reverse-phase high-performance liquid chromatography, which included pre-column derivation with orthophthaladehyde detection, showed varied increased levels of excitatory neurotransmitters, including aspartate and glutamate, and inhibitory neurotransmitters, including glycine and Y-aminobutyric acid, in the corpus striatum after treatment with moschus alone, diazepam alone, or a combination of both. Compared with the diazepam group, aspartate levels significantly decreased at 30 and 60-105 minutes after combined treatment with moschus, while glutamate significantly increased at 45 and 75-105 minutes, glycine levels significantly increased at 105 minutes, and γ-aminobutyric acid increased at 30 and 75-105 minutes. These findings suggested that moschus increased the inhibition effects of diazepam on the brain.

Effect of Diazepam on the Contents of Amino Acids and Free Radical during Ischemia/Reperfusion Injury

The protective effect and mechanism of diazepam on ischemia neurons during cerebral ischemia and reperfusion were studied. Sixty three Wistar rats were divided randomly into nine groups: control group , ischemia groups including subgroups of is3h, is3 h/rep1 h, is3 h/rep2 h, is3 h/rep3 h, diazepam treated groups , including subgroups of is3 h, is3 h/rep1 h, is3 h/rep2 h, is3 h/rep3 h with Zea longa's animal model of middle cerebral artery occlusion. The comparison between the ischemia group and diazepam treated group showed that diazepam could obviously decrease the production of glutamate, asparate, MDA and increase the synthesis and release of GABA, SOD and GSH PX. It was concluded that diazepam exerted its protective effects on neurons through complex mechanisms of regulating the synthesis and release of excitotary/inhibitory amino acids and free radicals.

Effects of Diazepam on Rat Cholinesterase andMembrane Stability of Red Cell in vitro

The results confirmed that diazepam inhibits the cholinesterase in rat serum,red cell,corpus striatum and diaphragm in vitro,that the higher the diazepam concentration,the stronger the cholinesterase inhibition,and that diazepam is a reversible inhibitor to acetylcholinesterase and diazepam has a stabilizing action on red cell membranes.The role of these effects of diazepam in the treatment of organic phosphate poisoning is discussed

Pharmacological Mechanisms of Diazepam in Fish： Effect on Growth

The present work aimed to demonstrate that the dietary content of the drug diazepam, a common benzodiazepine, regulates many aspect of feed efficiency of the Nile tilapia Oreochromis niloticus a favorite in fish production, and also to test introducing a simple new model in the investigation of the biological mechanisms of drug addiction. Diazepam was added to the basic diet at different levels （1.5, 3.0, 5.0, 7.0, 8.0, 9.0, 10.0, 12.0, 14.0 and 16.0 mg/100 g diet）. The experiments lasted for twelve weeks. The results obtained suggested that most of the diazepam doses were able to stimulate the growth parameters of O. niloticus, especially at 12.0 mg/100 gdiet. At some selected doses, diazepam reduces AChE specific activities in the tilapia brain and the inhibition was higher at the 12.0 mg dose. The clearance of diazepam in fish muscles and skin with lapse of time indicated that the fish treatment poses no health risk to the consumer. The recommended dose is the 12 mg DZP/100 mg diet. Finally, tilapia can be used as a new powerful model for the study of fish growth, which could provide insights into the mechanisms of drug addiction.

Diazepam reduces synaptic GABA type areceptor availability via multiple trafficking mechanisms

OBJECTIVE Investigate the effects of diazepam(DZP) on γ2 subunit containing GABA type A receptor(GABA A R) trafficking.METHODS Immunofluorescence microscopy measured surface GABA A Rs and gephyrin in rat cortical neurons after 24 h exposure of 1.0 μmol·L^(-1) DZP.Biochemical studies of mice injected with 10 mg·kg^(-1) DZP vs vehicle were assessed for γ2 subunit and total gephyrin cortical levels 12 h post-injection.Ubiquitination of the γ2 subunit was studied by immunoprecipitation after 12 h of 1.0 μmol·L^(-1) DZP exposure.A γ2 subunit encoding an N terminal fluorogen-activating peptide and pH-sensitive green fluorescent protein(γ2 pH FAP) measured lysosomal targeting of γ2 containing GABA A Rs.RFP-gephyrin and γ2 pH FAP synaptic diffusion rates were examined using fluorescence recovery after photobleaching(FRAP).RESULTS Extrasynaptic levels of γ2 GABA A Rs decreased by 12.2%,while synaptic gephyrin S270 phosphorylation increased by 18.3% in DZP-treated neurons after 24 h compared to control(P<0.05).Dendritic levels of gephyrin were also reduced to 74.1% of control,while S270 phosphorylation was elevated by 25.2%(P<0.05;P<0.01).Mice 12 h post-DZP injection demonstrated a 12.7% and 26.1% decrease in total γ2 and gephyrin levels,respectively(P<0.05;P<0.01).12 h DZP treatment enhanced γ2 subunit ubiquitination 1.13-fold relative to control(P<0.05).Internalized γ2 pH FAP GABA A Rs associated with lysosomes was 8.0% higher in neurons treated with 12-16 h DZP compared to control.Pilot FRAP experiments suggest gephyrin and γ2 have increased mobility and turnover at synapses following DZP.CONCLUSION DZP treatment decreases γ2 GABA A R levels and gephyrin scaffolding function after one day of exposure,which may contribute to the formation of DZP tolerance.

Effect of Cyclodextrin Complexation on the Aqueous Solubility of Diazepam and Nitrazepam:Phase-Solubility Analysis,Thermodynamic Properties

The solubility enhancement of diazepam and nitrazepam in water was analyzed depending on temperature and amount of α-cyclodextrin ( α-CD), β-cyclodextrin (β-CD) and 2-hydroxypropyl-β-cyclodextrin (2-HP-β-CD). The interactions of drug-cyclodextrin in solution were investigated by the phase-solubility analysis. Diazepam (nitrazepam) content in aqueous complexation medium was analyzed UV spectrophotometrically. Classical solubility data were used to derive apparent stability constants (K1:1) which were used to derive thermodynamic parameters for the diazepam (nitrazepam)-cyclodextrin complexes. Since all phase solubility plots were of AL–types, and calculated Slopes after linear regression analysis were found to be less than 1, it could be assumed that stoichiometry of the formed binary systems was 1:1. According to the calculated K1:1 values, the stability of the complexes of diazepam and nitrazepam with a-CD, β-CD and 2-HP-β-CD varies as follows: 2-HP-β-CD > β-CD > β-CD. The a-CD has higher affinity for dissolving nitrazepam compared to diazepam. While all parameters lead to an improvement in solubility, the largest effect was obtained for guest-host complexation with 2-HP-β-CD. The solubility of diazepam and nitrazepam in water increased 93.02 times and 64.23 times, respectively, in the presence of 40% (w/w) 2-HP-β-CD, at 25°C. Solubility data for diazepam and nitrazepam in aqueous 2-HP-b-CD were used to derive thermodynamic parameters, ΔG° at 298 K = –14.43 kJ·mol–1, ΔH° = 0.79 kJ·mol–1, ΔS° at 298 K = 51.17 J·mol–1·K–1 and ΔG° at 298 K = –13.43 kJ·mol–1, ΔH° = 2.38 kJ·mol–1, ΔS° at 298 K = 53.01 J·mol–1·K–1, respectively. Formation of inclusion complexes substantially increases the water solubility of diazepam and nitrazepam. Diazepam and nitrazepam dissolution thermodynamics in aqueous 2-HP-β-CD were characterized by spontaneous and endothermic dissolution and hydrophobic interactions.

Diazepam during endoscopic submucosal dissection of gastric epithelial neoplasias

AIM:To investigate risk factors and adverse events related to high-dose diazepam administration during endoscopic submucosal dissection for gastric neoplasias.METHODS:Between February 2002 and December 2009,a total of 286 patients with gastric epithelial neoplasia underwent endoscopic submucosal dissection in our hospital.To achieve moderate sedation,5-7.5 mg of diazepam was administered intravenously by non-anesthesiologists.Intermittent additional administration of 2.5-5 mg diazepam was performed if uncontrollable body movement of the patient was observed.All patients were classified into groups based on the required diazepam dose:low-dose (≤ 17.5 mg,n=252) and high-dose (> 17.5 mg,n=79).RESULTS:Differences between the low-and highdose diazepam groups were observed in lifetime alcohol consumption (0.30 ± 0.48 vs 0.44 ± 0.52 tons,P=0.032),body weight (58.4 ± 10.3 vs 62.0 ± 9.9 kg,P=0.006),tumor size (15 ± 10 vs 23 ± 18 mm,P < 0.001),lesion location (P < 0.001) and the presence of ulcerative findings (14/238 vs 18/61,P < 0.001).Multivariate analysis identified all five variables as independently related to required diazepam dosage.In terms of adverse reactions to diazepam administration,paradoxical excitement was significantly more frequent in the high-dose diazepam group (P < 0.001).CONCLUSION:Intermittent administration of diazepam enabled safe completion of gastric endoscopic submucosal dissection except in patients who were alcohol abusers or obese,or who showed complicated lesions.

Melatonin and Diazepam Affect Anxiety-Like and Depression-Like Behavior in Wistar Rats: Possible Interaction with Central GABA Neurotransmission

Recent studies have shown the importance of the GABA-ergic transmission in the pathophysiology of anxiety and depressive disorders in humans. Our present study aims to investigate the interaction of melatonin (MEL) with this system by exploring the effects of MEL with or without a facilitator of GABA-ergic neurotransmission, diazepam (DZ) on the levels of depression and anxiety in Wistar rats. For this purpose, different doses of MEL (2, 4 or 16 mg/kg) or DZ (2 mg/kg) are subchronically administered during 15 days. After pharmacological treatments, anxiety levels are evaluated in behavioral tests of Open Field (OFT) and elevated plus maze (EPM) and depression levels are evaluated by the forced swim test (FST). The results showed that MEL produces anxiolytic-like and antidepressant-like effects in a dose-dependent manner;the maximum effect was obtained at a dose of 16 mg/kg. However, a dose of 4 mg/kg is necessary to induce an effect. The effect of MEL and DZ reported in this study concerns selective modulation of behavioral anxiety and depression since locomotor activity assessed by the OFT and EPM was not affected. The subchronic injection of MEL at 4 mg/kg, DZ at 2 mg/kg or the two combined molecules also induces also anxiety-like and antidepressant-like behavior. In addition, a potentiating effect between MEL and DZ was observed. These effects suggest that psychopharmacological actions of MEL are due, at least in part, to its ability to improve the central GABA-ergic transmission.

PROPHYLAXIS OF INTERMITTENT DIAZEPAM IN CHILDREN WITH FEBRILE CONVULSIONS（FC）

The purpose of this study was to limit intermittent diazcpam prophyiaxis of children with FC to the patients who haye the second recurrence.A series of 156 children with FC received prophylactic treatment.During 1￣5 years for Follow-up (average,2 years and 10 months),28 cases of prophylactic group suffered recurrence of FC 48 times. 54 of 126 cases in control group suffered it 108 times. The difference in case number and recurrent rate between the prophylactic and control groups was highly significant (P<0. 01). Diazepam was found tO be considerably effective in reducing the risk of recurrence of FC.

Intranasal Delivery of Two Benzodiazepines, Midazolam and Diazepam, by a Microemulsion System

Nasal application of benzodiazepines might be an alternative to intravenous administration in acute clinical situations such as seizures emergencies. However, irritation and pain as well as symptoms like teary eyes, dizziness, discomfort, nasal drainage and bad taste usually accompany subject received midazolam and diazepam via the nasal route. The purpose of this study was to evaluate the use of a new alcohol-free microemulsion system as a carrier for diazepam or midazolam given intranasally. Midazolam (base) or diazepam was solubilized in the microemulsion to obtain a high drug concentration of 25 mg/g (2.5% by weight), to provide 2.5 mg drug in 100 μl spray (d ≈ 1.00 g/ml). The nasal absorption of both drugs from the same microemulsion formulation (containing 20% aqueous phase) was found to be fairly rapid after administration of 0.4 mg/kg to rabbits. The absolute bioavailability of diazepam after intranasal administration using this formulation was 33.45% ± 12.36% and the tmax was 18.33 ± 23.09 min, which was twice longer than the tmax obtained after midazolam administration, 9.25 ± 6.75 min. The pharmacokinetic parameters of midazolam in W/O (20% water) microemulsion and their comparison with midazolam in O/W (50% water) microemulsion have shown that both formulations resulted in a relatively short time to reach the peak plasma level (tmax), that is, 9.25 ± 6.75 min and 6.75 ± 5.67 min, respectively. However, the peak plasma levels (Cmax) and the absolute bioavailability (FA) of midazolam were significantly higher after administration of the W/O formulation than those obtained after application of O/W formulation, i.e., 46.62 ± 17.38 μg/ml vs. 15.44 ± 4.00 μg/ml, and 35.19% ± 11.83% vs. 19.83% ± 16.32%, respectively. Our results suggest that the new microemulsion system may be useful for getting rapid-onset of midazolam and diazepam following intranasal administration, resulting in reasonable peak plasma levels and bioavailability, but most importantly, providing a high measure of tolerability and comfort.

Maintenance time of sedative effects after an intravenous infusion of diazepam: A guide for endoscopy using diazepam

AIM: To examine whether the sedative effects assessed by psychomotor tests would depend on the cytochrome P450 (CYP ) 2C19 genotypes after an infusion regimen of diazepam commonly used forgastrointestinal endoscopy in Japan. METHODS: Fifteen healthy Japanese volunteers consisting of three different CYP2C19 genotype groups underwent a critical ? icker fusion test, an eye movement analysis and a postural sway test as a test for physical sedative effects, and a visual analog scale (VAS) symptom assessment method as a test for mental sedative effects during the 336 h period after the intravenous infusion of diazepam (5 mg). RESULTS: The physical sedative effects assessed by the critical flicker test continued for 1 h (t values of 5 min, 30 min and 60 min later: 4.35, 5.00 and 3.19, respectively) and those by the moving radial area of a postural sway test continued for 3 h (t values of 5 h, 30 h, 60 min and 3 h later: -4.05, -3.42, -2.17 and -2.58, respectively), which changed significantly compared with the baseline level before infusion (P < 0.05). On the other hand, the mental sedative effects by the VAS method improved within 1 h. The CYP2C19 genotype-dependent differences in the postinfusion sedative effects were not observed in any of the four psychomotor function tests. CONCLUSION: With the psychomotor tests, the objective sedative effects of diazepam continued for 1 h to 3 h irrespective of CYP2C19 genotype status and the subjective sedative symptoms improved within 1 h. Up to 3 h of clinical care appears to be required after the infusion of diazepam, although patients feel subjectively improved.

Propofol combined with diazepam synergistically potentiates the GABA-activated chloride current in rat sensory neurons

To investigate the effect of propofol combined with diazepam on the γ-aminobutyric acid (GABA)-activated chloride current (I GABA ) evoked in rat sensory neurons Methods Whole cell patch clamp recordings were made from cultured rat dorsal root ganglionic neurons GABA (3*!μmol/L) was applied by pressure ejection The anesthetics were dissolved in the external solution and given by the “Y-tube' method Results Co-application of propofol (0 3-3*!μmol/L) and diazepam (100*!nmol/L) potentiated the I GABA which was significantly larger than the sum of that potentiated by drug alone Diazepam (100*!nmol/L) shifted the concentration-response curve for the I GABA potentiation induced by propofol to the left in a parallel fashion The EC 50 value for propofol was decreased by diazepam from 7 6±1 8 *!μmol/L to 3 9±1 1*!μmol/L (n=9) Conclusions Our results suggest that propofol combined with diazepam synergistically potentiates the I GABA Diazepam-induced increase in the apparent binding affinity of propofol for the GABA A receptors is likely responsible for a clinical synergistic hypnotic action during co-application with propofol and diazepam

A Preliminary Gas Chromatography‑Mass Spectrometry‑Based Metabolomics Study of Rats Ingested Diazepam or Clonazepam

Drug-facilitated sexual assault(DFSA)is a sexual act in which the victim is unable to give or rescind consent due to alcohol or drug intoxication,which involved the abuse of benzodiazepines around the world.Conventional techniques used for the analysis of benzodiazepines have the limitation of short detection time window due to the rapid metabolism of these drugs in body.This study aimed to investigate the characteristic changes of metabolites in the blood of rats after ingesting diazepam/clonazepam through a gas chromatography-mass spectrometry-based metabolomics method,allowing the indirect reveal of the rats ingested diazepam/clonazepam.First,we found that diazepam and clonazepam in the blood of rats could not be detected by liquid chromatography-tandem mass spectrometry after 48 h of ingestion.Then,orthogonal partial least squares discrimination analysis regression models were,respectively,constructed to determine whether the rats ingested diazepam/clonazepam after 48 h.The results showed that 5 metabolites were found to be associated with diazepam exposure,and 7 metabolites were found to be associated with clonazepam exposure,which may be characterization for the evaluation of digestion of diazepam and clonazepam in rat.

地西泮在模拟养殖环境中的含量变化及累积特征

为探究地西泮(Diazepam,DZP)在模拟养殖环境中的降解特点及累积特征,设置2个浓度胁迫组(A、C组),并在2个浓度下添加蜈蚣草(Pteris vittata)作对照组(B、D组),共4个试验组;分析水体、底泥和蜈蚣草中DZP浓度随时间的变化特点,探讨蜈蚣草和底泥对水体中DZP的累积特征。结果表明,给药后4组水体中DZP的初始质量浓度分别为A:(0.118±0.002)μg·L^(-1)、B:(0.117±0.004)μg·L^(-1)、C:(1.141±0.078)μg·L^(-1)和D:(1.142±0.039)μg·L^(-1),给药后第768小时水体中DZP质量浓度下降了29.71%~40.17%;DZP降解半衰期介于65.29~139.11 d。4组底泥中DZP质量分数随时间变化逐渐上升,给药768 h后4组底泥中DZP质量分数分别达到初始质量分数的17.99倍(1.384μg·kg^(-1))、14.81倍(0.918μg·kg^(-1))、4.77倍(7.848μg·kg^(-1))和5.30倍(7.763μg·kg^(-1)),富集系数介于9.79~18.80;B、D组蜈蚣草中DZP浓度峰值出现在给药后第216小时。蜈蚣草和底泥对水体中的DZP具有一定的吸附和富集作用,可明显缩短高浓度DZP在水中降解的半衰期,在低浓度DZP水体中添加蜈蚣草可抑制底泥对DZP的富集。

高效液相色谱-质谱法测定淡水鱼中地西泮

建立高效液相色谱-质谱法测定淡水鱼中地西泮。样品用乙腈提取,采用Florisil填料和氨基填料净化,当Florisil填料和氨基填料的质量比为2∶3时,能够有效地吸附淡水鱼基质中的脂肪、脂质和蛋白质等杂质,降低基质效应,提高质谱响应的信噪比。地西泮的质量浓度在0.05~20 ng/mL范围内与色谱峰面积线性关系良好,相关系数为0.9996,检出限为0.15μg/kg,定量限为0.5μg/kg。空白样品加标回收率为84.2%~95.5%,测定结果的相对标准偏差为5.25%~6.83%(n=6)。该方法快速准确,基质效应小,能够满足水产品中地西泮的测定要求。

液相色谱-串联质谱法检测淡水养殖环境中地西泮及其代谢物

建立了养殖水、植物、沉积物和鲫鱼中地西泮及其代谢物(去甲西泮、奥沙西泮和替马西泮)的液相色谱-串联质谱检测方法。养殖水经0.45µm玻璃纤维膜过滤,用氨水调至pH 8.0后通过反相固相萃取柱(HLB)净化;植物和沉积物经盐析提取后,通过QuEChERS方法净化上样;鲫鱼样品经盐析提取后,通过固相萃取柱(Prime HLB)净化上样。采用Luna C_(18)柱(100 mm×2.1 mm,1.6µm)进行分离,多反应监测(MRM)模式进行测定,外标法定量。结果表明:地西泮及其3种代谢物在0.1~10µg/L质量浓度范围内呈良好的线性关系(r^(2)>0.999)。4种目标物在养殖水体中的检出限为1.0~1.2 ng/L,定量下限为5.0~6.0 ng/L;在植物、沉积物和鲫鱼中的检出限为0.03~0.10µg/kg,定量下限为0.10~0.25µg/kg。在低、中、高3个加标水平下,地西泮及其代谢物的回收率为83.4%~106%,相对标准偏差(RSD,n=6)为1.2%~9.4%。该方法成功应用于养殖水、沉积物、植物和鲫鱼样品中地西泮及其代谢物的检测,能为地西泮污染追查提供检测方法支撑。

QuEChERS/HPLC-MS/MS法测定渔用饲料及饵料中地西泮及其代谢物

研究建立了一种同时测定渔用饲料及饵料中地西泮及其代谢物的分散型固相萃取(QuEChERS)/高效液相色谱-串联质谱(HPLC-MS/MS)的方法。样品采用乙酸乙酯提取,提取液经50 mg C18和PSA分散固相萃取净化,40℃下氮吹浓缩近干,加入80%乙腈1.0 mL复溶,过滤后采用HPLC-MS/MS法进行测定。目标物采用飞诺美Kinetex C18系列色谱柱(100 mm×2.1 mm,2.6μm)分离,以0.1%体积比甲酸水溶液和乙腈为流动相梯度洗脱。电喷雾离子源(ESI+)、选择反应监测(SRM)模式下进行测定,基质匹配内标法定性定量。结果显示,该方法在0.5~50.0μg/kg范围内线性关系良好,相关系数均大于0.999,检出限和定量下限分别为0.5μg/kg和1.0μg/kg。在3个不同的添加水平下,地西泮及其代谢物的平均回收率为82.3%~109.4%,相对标准偏差为3.62%~6.33%。研究表明,该方法具有快速、准确度高与灵敏度高的优点,可满足渔用饲料及饵料中地西泮及其代谢物的残留检测需求。

液相色谱-串联质谱法测定鱼饵中地西泮的不确定度评定

试验为评定液相色谱-串联质谱法测定鱼饵中地西泮含量的不确定度。试验通过建立数学模型分析不同来源的不确定度,评定各不确定度分量。结果显示,测量结果的相对标准不确定度为4.55%,相对扩展的不确定度为9.10%,样品中地西泮含量为(23.821±2.168)μg/kg(K=2)。研究表明,鱼饵中地西泮含量的测量不确定度主要源于试样重复测量、标准溶液配制和试样称量。

地西泮通过let-7a-5p/MYD88轴抑制LPS诱导的细胞焦亡和炎症从而缓解小鼠肺纤维化

目的探讨地西泮(Dia)如何通过介导let-7a-5p与MYD88之间的靶向调控作用,来减轻由脂多糖(LPS)诱导的细胞焦亡和炎症反应,从而缓解肺纤维化的进程。方法MRC-5细胞分为正常对照组(NC)、LPS组、LPS+Dia组,LPS+Dia+let-7a-5p mimic组、LPS+Dia+let-7a-5p mimic+pc-DNA-MYD88组。RT-qPCR检测let-7a-5p和MYD88的表达。ELISA检测炎症因子的表达,Western blotting检测纤维化和焦亡相关蛋白的表达。C57BL/6小鼠随机分为NC组、LPS组、LPS+Dia、LPS+Dia+let-7a-5p mimic组、LPS+Dia+ST2825(MYD88抑制剂)组、LPS+Dia+let-7a-5p mimic+pc-DNA-MYD88组。Masson染色观察小鼠肺纤维化,免疫荧光检测α-SMA的表达。结果与NC组比较,LPS组let-7a-5p的表达下调(P<0.01),MYD88的表达上调(P<0.01)。炎症相关因子IL-4、IL-6、TGF-β和TNF-α的浓度均升高(P<0.001),此外,纤维化相关蛋白Col-Ⅰ、Col-Ⅲ、α-SMA和细胞焦亡相关蛋白NLRP3、Caspase-1、ASC、GSDMD-N的表达量也升高(P<0.05)。与LPS组相比,LPS+Dia组有效抑制了炎症相关因子、纤维化相关蛋白、细胞焦亡相关蛋白的表达。动物实验中,与LPS组相比,LPS+Dia组小鼠肺组织纤维化程度降低,α-SMA相对荧光密度降低(P<0.05)。此外,与LPS+Dia组相比,LPS+Dia+let-7a-5p mimic组或LPS+Dia+ST2825的处理进一步促进了Dia的作用(P<0.05)。而过表达MYD88又削弱了let-7a-5p mimic对Dia的作用(P<0.05)。结论Dia可以通过上调let-7a-5p的表达负调控MYD88抑制LPS诱导的细胞焦亡和炎症反应从而缓解肺纤维化。