1.Effect of α-lipoic acid on cognitive function after cardiopulmonary bypass in diabetic rats
Guangming YIN ; Shanglong YAO ; Huaqing SHU
Chinese Journal of Anesthesiology 2012;(12):1437-1440
Objective To evaluate the effect of α-lipoic acid on the cognitive function after cardiopulmonary bypass (CPB) in diabetic rats.Methods Health adult male Sprague-Dawley rats,weighing 400-500 g,aged 16-22 weeks,were used in this study.Diabetes mellitus was induced by intraperitoneal streptozocin 60 mg/kg and confirmed by blood glucose≥ 16.7 mmol/L.Thirty-two diabetic rats were randomly divided into 2 groups (n =16 each):diabetes mellitus group (group D) and α-lipoi cacid group (group L).In group L,α-lipoic acid 30 mg/kg was intraperitoneally injected once a day for 7 consecutive days starting from 6th week after induction of diabetes mellitus.While the equal volume of normal saline was given instead in group D.The two groups underwent CPB after the last administration.Before induction of diabetes mellitus,on 5th week after induction of diabetes mellitus,before CPB,at the end of CPB,and on 3 and 5 days after termination of CPB,10 rats were chosen from each group and venous blood samples were collected for determination of plasma TNF-α and IL-10 concentrations.Ten rats in each group were chosen for detection of cognitive function before induction of diabetes mellitus,before CPB and 5 days after termination of CPB.The rats were then sacrificed and hippocampi were isolated for measurement of NF-κB activity.Results Compare with group D,the plasma TNF-α concentration,times of electric shock and activity of NF-κB in hippocampal tissues were significantly decreased and the plasma IL-10 concentration was increased in group L (P < 0.05 or 0.01).Conclusion α-lipoic acid can improve the cognitive function after CPB in diabetic rats and inhibition of activation of NF-κB in hippocampal neurons is involved in the mechanism.
2.Determination of the Content of Astragaloside Ⅳ in Qi Zao Granules By HPLC-ELSD
Huaqing LIN ; Hong DENG ; Shu ZHANG
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(02):-
Objective To determine the content of astragaloside Ⅳin Qi Zao Granules.Methods Equipped with the evaporative light scattering detector (ELSD), a RP-HPLC method was established. The chromatographic column was C18 and the column temperature was at 30 ℃.The drift tube temperature was at 40 ℃and the evaporative gas was nitrogen with pressure of 3.5 bar.The mobile phase was acetonitrile-water(30∶70) and the flow rate was 1mL/min. Results This method showed a good linear relationship in the range of 2.51~50.2 ?g astragaloside Ⅳ. The average recovery was 98.13 %with RSD being 1.28 %. Conclusion The method is sensitive and accurate and can be used for the quality control of Qi Zao Granules.
3.Pharmacokinetics of different concentration of ropivacaine following lumbar plexus combined with sciatic nerve block
Huaqing SHU ; Xiaoming ZHANG ; Shanglong YAO
Chinese Journal of Anesthesiology 1997;0(11):-
Objective To evaluate the pharmacokinetics of ropivacaine following lumbar plexus combined with sciatic nerve block for knee arthroscopy. Methods After obtaining written informed consent 16 ASAⅠorⅡpatients of both sexes (8 males ,8 females) scheduled for unilateral knee arthroscopy under lumbar plexus combined with sciatic nerve block with ropivacaine were randomly divided into 2 groups (n = 8 each) : group A received 0.5% ropivacaine 30 ml (15.0 mg) and group B received 0.75% ropivacaine 30 ml (22.5 mg) . Blood samples were taken from radial artery immediately after and at 5, 10, 15, 30, 45, 60, 120 and 180 min after drug administration for determination of plasma ropivacaine concentration by HPLC. The pharmacokinetic parameters were calculated using 3p97 computer program. Results The two groups were comparable with respect to sex ratio (M/F), age, body weight, height, duration of operation and amount of fluid infused. The main pharmacokinetic parameters in group A and B were: Cmax(1.4?0.3) mg?L-1 and (2.7?0.9) mg?L-1;AUC (3.88?0.28) mg?L-1?h-1 and (7.13?0.65) mg?L-1?h-1;t1/2?(0.44?0.19)h and (0.60?0.34)h; t1/2?(3.4?0.4)h and (2.4?0.5)h. The Cmax in group B was significantly higher than that in group A. The Cmax of ropivacaine reached 3.57 mg?L-1 in group B. No patient developed central nervous system or cardiac toxicity.Conclusion The plasma concentration versus time curve is fitted to two-compartment pharmacokinetic model. Lumbar plexus combined with sciatic nerve block with 0.5% ropivacaine 30 ml is safer.
4.Effects of mechanical stretch on expression of pentraxin-3 mRNA and protein in A549 cells
Huaqing SHU ; Qingping WU ; Lei YANG ; Hong ZHANG ; Shanglong YAO
Chinese Journal of Anesthesiology 2008;28(11):1023-1025
Objective To evaluate the effects of mechanical stretch on pentraxin-3(PTX-3)mRNA and protein expression in human alveolar epithelial cells (A549 cells).Methods The human lung epithelial adenocarcinoma cells A549(A549 cells)were purchased from cell biology laboratory,Tongji Medical College,Huazhong University of Science and Technology.The cultured A549 cells were inoculated on collagen Ⅰ BioFlex plates and divided into 5 groups(n=3 wells each):group Ⅰ normal control;groupⅡsham mechanical stretch;group Ⅲ mechanical stretch;groupⅣsiRNA and groupⅤ siRNA+mechanical stretch.In group Ⅲ the cells underwent square cyclic mechanical stretch for 4 h using the Flexercell Systcm.In group Ⅳ the cells were transfected with chemosynthetic PTX-3 specific siRNA by RNAi technique.In group Ⅴ at 24 h after being transfected with PTX-3 siRNA the cells underwent mechanical stretch for 4 h.In groupⅡ mechanical stretch of the cells were prevented by Flexstep.The expression of PTX-3 mRNA in the cells was detected by real-time PCR and the expression of PTX-3 protein in the culture media was determined by Western blotting.Apoptosis of the cells was measured bv flow cytometry(Beeten-Dickinson,USA).Results PTX-3 mRNA and protein expression was signlficantly up-regulated by mechanical stretch in groupⅢand decreased by transfection with siRNA in group Ⅳand Ⅴ as compared with group Ⅰ andⅡ(P<0.05 or 0.01).The apoptosis ratio was significantly higher in group Ⅲ and Ⅴ than in groupⅡ and was significantly lower in group Ⅴ thanin group Ⅲ(P<0.01).Conclusion Mechanical stretch can up-regulate PTX-3 mRNA expression in A549 cells.
5.Effects of mechanical stretch on the expression of human beta-defensin-3 in alveolar epithelial cells elicited by interferon- gamma
Ping GUI ; Qingping WU ; Shanglong YAO ; Huaqing SHU
Chinese Journal of Emergency Medicine 2008;17(6):606-609
Objective To study the effect of mechanical stretch on the expression of human beta-defensin-3 (HBD-3) in alveolar epithelial cells(A549 cells) elicited by interferon-gamma(IFN-γ) and to investigate the role of HBD-3 in the pathogenesis of ventilator-associated pneumonia (VAP) . Method A549 cells cultured in vitro were treated with mechanical stretch (group S), 10 ng/ml IFN-γ (group I) ,and 10 ng/ml IFN-γ with mechanical stretch (group IS), respectively. Cells without treatment served as controls (group C). Cells were stretched by 20% amplitude of stretch at 30 cycles/mm by Flexercell-4000[TM]Unit for 2 h, 4 h, and 6 hours. The HBD-3 mRNA expression was determined by real-time RT-PCR after treatment. After 6 hours, treatment, cells were cultured for 24 hours and the expression of HBD-3 was examined by laser scanning confocal microscope. The experimental data were statistically analyzed by using one-way ANOVA analysis and q-test. Results The expression of HBD-3 mRNA in A549 cells could not significantly be changed by mechanical stretch alone. Compared with group C,the HBD-3 mRNA expression after treatment with 10 ng/ml IFN-γ for 2 hours,4 hours and 6 hours increased significantly by (2.63 C,the HBD-3 mRNA expression after treatment with 10 ng/ml IFN-γ and mechanical stretch for 2 hours,4 hours and 6 hours increased by (1.54 were significantly lower than those in group I (P < 0.01). The HBD-3 expression in group IS after mechanical stretch for 6 significantly different from than in group C. Conclusions Mechanical stretch can significantly suppress the up-regulation of HBD-3 in alveolar epithelial cells elicited by IFN-γ, and this may be one of the explaina-tions that patients under mechanical ventilatiori(MV) have a higher risk of VAP.
6.Application progress of point-of-care ultrasound monitoring inferior vena cava in volume management of critically ill patients
Xuehui GAO ; Huaqing SHU ; Yuan YU ; Xiaobo YANG ; You SHANG
Chinese Critical Care Medicine 2021;33(11):1379-1383
Determining whether patients have volume-responsiveness is one of the frequently asked questions in the intensive care unit, especially in shock patients. Evaluating the volume status and volume responsiveness can help clinical medical staff accurately grasp the patient's cardiac preload, guide reasonable volume management, and help improve patient prognosis. Therefore, many non-invasive and invasive methods have been proposed to evaluate volume responsiveness. Inferior vena cava ultrasound has been widely used to guide the fluid management of critically ill patients due to its simplicity, non-invasiveness, and good repeatability. This article reviews the clinical applications of inferior vena cava ultrasound in fluid management of critically ill patients, so as to provide a reference for circulatory management of critically ill patients.
8.ATPR alleviates lipopolysaccharide-induced acute liver injury in mice by promoting autophagy
Chuanlin SHU ; Xiaorui SHI ; Rumeng ZHU ; Qing ZHOU ; Yuan WANG ; Yi WANG ; Huaqing ZHU
Acta Universitatis Medicinalis Anhui 2024;59(2):200-206
Objective To investigate the effect of4-amino-2-trifluoromethyl-phenyl retinate(ATPR)on acute liver injury induced by lipopolysaccharide(LPS)in C57BL/6 mice and its related mechanism.Methods Fifteen 6-week-old male C57BL/6 strain mice were randomly divided into normal group,model group and ATPR group,with 5 mice in each group.Mice in the ATPR group were intraperitoneally injected with ATPR(15 mg/kg·d),and normal group and model group were given solvent.After continuous administration for one week,model group and ATPR group were intraperitoneally injected with LPS(6 mg/kg),and all mice were sacrificed 6 hours later.The contents of Alanine aminotransferase(ALT)and Aspartate aminotransferase(AST)in serum of mice were detec-ted.The mRNA levels of Interleukin-6(IL-6)and Tumor necrosis factor-alpha(TNF-α)were detected by qPCR.Hematoxylin-eosin(H&E)staining was used to observe the histopathological changes of liver in mice.The ultra-structural changes of mouse hepatocytes were observed by Transmission electron microscope(TEM).The expres-sion levels of mitochondrial damage-related proteins FUNDC1 and OPA1 and autophagy related proteins LC3B,P62,Beclin1 and ATG5 were detected by Western blot.Results Compared with the normal group,the content of ALT and AST in serum and the mRNA levels of IL-6 and TNF-α in liver tissue increased in the model group,and the changes were reversed in the ATPR group.H&E staining showed that the hepatic lobule structure was normal in the normal group,the hepatic cords were arranged radially,there was no hyperemia and inflammatory cell infiltra-tion,and the hepatocyte boundary was clear.In the model group,the intercellular space of liver was enlarged,the arrangement of hepatic cords was disordered,and inflammatory cells infiltrated.In the ATPR group,the intercellu-lar space of liver and the structure of hepatic cords were restored,and the inflammatory cell infiltration was less.TEM showed that the damaged mitochondria and lipid droplet accumulation in the hepatocytes of mice in the model group were compared with that in the normal group,and the morphology and quantity of mitochondria and lipid droplet in the hepatocytes of mice in the ATPR group tended to be normal.Western blot showed that compared with the normal group,the expression of FUNDC1 protein in the liver tissues of mice in the model group increased,the expression of OPA1 protein decreased,the ratio of LC3B Ⅱ to LC3B Ⅰ decreased,the expression of P62 protein in-creased,the expression of Beclin1 and ATG5 protein decreased,and the above changes were reversed in the ATPR group.Conclusion ATPR alleviates acute liver injury induced by lipopolysaccharide in mice by promoting autoph-agy.
9.ATRA alleviates lipopolysaccharide -induced acute myocardial injury in mice by inhibiting Ferroptosis
Zhangyong Dan ; Chuanlin Shu ; Xiaorui Shi ; Qing Zhou ; Yuan Wang ; Mengfei Ren ; Yi3 Wang ; Huaqing Zhu
Acta Universitatis Medicinalis Anhui 2023;58(3):373-378
Objective :
To investigate the effect and mechanism of all-trans retinoic acid (ATRA) on lipopolysaccharide (LPS) -induced acute myocardial injury in C57BL/6 mice.
Methods :
Male mice of C57BL/6 strain were randomly divided into normal group,model group,ATRA group,and ferrostatin-1 group.Mice in the ATRA group were injected intraperitoneally with ATRA 15mg / (kg · d) ,ferrostatin-1 group received ferrostatin-1 2 mg / ( kg · d) ,the normal group and the model group were given solvent.After one week of continuous administration,the model group,ATRA group ,and ferrostatin-1 group were intraperitoneally injected with LPS 6 mg / kg. All mice were sacrificed after 6 hours.The contents of malondialdehyde ( MDA) and glutathione ( GSH) in serum of mice were detected. qPCR was used to detect mRNA levels of interleukin-6 ( IL-6 ) and tumor necrosis factor-alpha (TNF-α) in heart tissue.Hematoxylin-eosin ( HE) staining was used to observe the changes of heart tissue in mice.Transmission electron microscopy (TEM) was used to observe the structure of mouse myocardial mitochondria.Western blot was used to detect the expression of ferroptosis markers glutathione peroxidase 4 ( GPX4) ,ferritin heavy chain 1 ( FTH1 ) ,Solute carrier family 7 member 11 ( SLC7A11 ) ,acyl-CoA synthetase long-chain family member 4 (ACSL4) and related regulatory proteins,Nuclear factor erythroid 2-related factor 2 ( NRF2 ) ,kelchlike ECH-associated protein 1 (KEAP1) .
Results:
Compared with the normal group,the MDA content in the serum of the model group increased and the GSH content decreased,the above changes were reversed in the ATRA group as well as in the ferrostatin-1 group.Compared with normal group,the mRNA levels of IL-6 and TNF-α in the heart tissue of model group increased steeply,the above changes were relieved in the ATRA group and the ferrostatin-1 group.There was no significant difference in HE staining of myocardial tissue among the groups of mice. Compared with the normal group,myocardial mitochondria in the model group showed the phenomenon of cristaereduction or disappearance under TEM,while myocardial mitochondrial injury was alleviated in the ATRA group and the ferrostatin-1 group.Western blot showed that GPX4,FTH1,SLC7A11,and NRF2 expression were reduced in the myocardial tissue of mice in the model group compared with the normal group,ACSL4 and KEAP1 expression increased.The above changes were reversed in the ATRA group as well as in the ferrostatin-1 group.
Conclusion
ATRA alleviates lipopolysaccharide-induced acute myocardial injury in mice by inhibiting ferroptosis.