1.Role of spinal HCN channels in dexmedetomidine-produced antinociceptic effect: in vivo and in vitro experiments
Yingcong YANG ; Xiangdong CHEN ; Zhongyuan XIA
Chinese Journal of Anesthesiology 2015;35(9):1096-1100
Objective To investigate the role of spinal hyperpolarization-activated cyclic nucleotide-gated (HCN) channels in dexmedetomidine-produced antinociceptic effect.Methods In vivo experiment Thirty wild type C57BL/6J mice and 30 HCN1 gene knockout (HCN1-/-) mice, aged 2-3 months, weighing 19-25 g, were randomly divided into 5 groups (n=6 each) using a random number table: control group (group C), and dexmedetomidine 10, 20, 30 and 40 μg/kg groups (Dex10, Dex20,Dex30 and Dex40 groups).In Dex10, Dex20, Dex30 and Dex40 groups, dexmedetomidine 10, 20, 30 and 40 μg/kg were intraperitoneally injected, respectively.The equal volume of normal saline was given in group C.Before dexmedetomidine administration, and at 15, 30, 45, 60, 75, 90, 105 and 120 min after dexmedetomidine administration, tail flick latency to a thermal nociceptive stimulus was measured, and the percentage of the maximum possible effect (MPE%) was calculated.In vitro experiment HCN1 and HCN2 plasmids and green fluorescent plasmids were transfected into HEK293 cells with liposome 2000.At 24-48 h after transfection, HCN1 and HCN2 channel currents were recorded using whole-cell patch clamp technique.HCN channel currents were recorded as baseline value after rupture of membrane.HEK293 cells were perfused with the extracellular fluid containing different concentrations of dexmedetomidine (0.1, 1.0 or 10.0 μmol/L).After the cells were perfused with dexmedetomidine 0.1 μmol/L for 5 min, HCN currents were recorded.The inhibition rate of currents were calculated.After washout, HCN currents were recorded after the cells were perfused with the next concentration for 5 min.The half-maximal activation voltage (V1/2) of HCN channels and the curve slope were recorded.The difference in V1/2 before and after administration (△V1/2) were calculated.Results Compared with group C, MPE% was significantly increased in Dex10 group-Dex40 group of wild type and HCN1-/-mice (P<0.05).Compared with Dex30 and Dex40 groups of wild type mice, MPE% was significantly decreased in Dex30 and Dex40 groups of HCN1-/-mice (P<0.05).There was no significant difference in MPE% between Dex10 group and Dex20 group of wild type and HCN1-/-mice (P>0.05).Compared with the baseline value, the currents and V1/2 of HCN1 and HCN2 channels in HEK293 cells were significantly decreased when the cells were perfused with dexmedetomidine 0.1, 1.0 and 10.0 μmol/L (P<0.05).Compared with the value when the cells were perfused with dexmedetomidine 0.1 μmol/L, the currents and V1/2 of HCN1 and HCN2 channels in HEK293 cells were significantly decreased, and inhibition rate of currents and △ V1/2were increased when perfused with dexmedetomidine 1.0 and 10.0 μmol/L (P<0.05).Compared with the value when the cells were perfused with dexmedetomidine 1.0 μmol/L, the currents and V1/2 of HCN1 and HCN2 channels in HEK293 cells were significantly decreased, and inhibition rate of currents and △ V1/2were increased when perfused with dexmedetomidine 10.0 μmol/L (P<0.05).There was no significant difference in the activation curve slope of HCN1 and HCN2 channel currents in HEK293 cells when the cells were perfused with dexmedetomidine 0.1, 1.0 and 10.0 μmol/L (P>0.05).Conclusion Dexmedetomidine-produced antinociceptic effect is likely related to the inhibition of spinal HCN channel opening.
2.Effects of propofol in TNF-induced apoptosis in endothelial cells derived from human umbilical vein
Tao LUO ; Zhongyuan XIA ; Xuejun CHEN
Chinese Journal of Anesthesiology 1995;0(10):-
Objective To investigate the protective effects of propofol in clinical relevant concentration on TNF-induced apoptosis in endothelial cells derived from human umbilical vein (HUVECs) .Methods The third and fourth passages of the cultured endothelial cells were divided into 5 groups. 0, 12.5, 25, 50 or 100 ?mol/L propofol was added to the cultured cells respectively and the cells were incubated for 30 min. Then TNF-? was added to the cultured cells (the final concentration of TNF-? was 2000 U.ml-1 ) which was incubated for 24h. Apoptosis was detected by using "terminal deoxynucleotidyl transferase (tdt )-mediated deoxyuridine triphosphate (dUTP )-biotin nick end-labelling" (TUNED . Morphologic changes were examined by means of electron microscope.Results Apoptosis expression was very high in control group (no propofol was added) . The process can be inhibited by pre-treatment with propofol (P
3.Pulmonary inflammatory pseudotumor:clinical characteristics and surgical treatment
Lixin SHAO ; Zhongyuan CHEN ; Jianbiao WU
China Oncology 2000;0(06):-
Purpose: To investigate the pathogenesis and clinical treatment of pulmonary inflammatory pseudotumor (PIP). Methods:To analyze the pathogenic characteristics and X-ray findings of 18 cases of PIP. Results:The rate of incidence in PIP is increasing in recent years .70 percent of the patients have infection of the respiratory system and is related to the abuse of large doses of antibiotics, there is little specificity in X-ray or CT so it is easy to confuse with malignant tumor of the lung .In pathologic sections of PIP, no malignant changes were seen, and only inflammatory cells, lymphocytes and plasm cells could be seen by light microscope. Conclusions:The antibiotic must be used properly in infection of the respiratory system and surgical treatment can be done in some asses to exclude tumor of the lung in diagnosis.
4.Mechanism of protective effect of Shen-Fu injection(参附注射液)on renal failure induced by intestinal ischemia-reperfusion in rats
Yuhong HE ; Chang CHEN ; Zhongyuan XIA
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2006;0(02):-
Objective To explore the effect of Shen-Fu injection(SFI,参附注射液) on expressions of heme oxygenase-1(HO-1) and inducible nitric oxide synthase(iNOS) in renal failure induced by intestinal ischemia-reperfusion injury(IRI) in rats and its possible mechanism in the protection of kidney.Methods The model of intestinal IRI was induced by clamping superior mesenteric artery(SMA) for 1 hour and then releasing the arterial clamp for 6 hours.Thirty-six male Wistar rats were randomly divided into three groups: IRI model group,SFI pretreatment group and sham operation group.In the SFI pretreatment group,10 ml/kg of SFI was pumped in at constant rate 30 minutes before the ischemia,the SMA was clumped for 1 hour and then released,while in the IRI model group,an equal volume of normal saline was pumped in continuously 30 minutes before the ischemia.The serum creatinine(SCr) and blood urea nitrogen(BUN) were observed respectively.Expressions and distributions of HO-1 and iNOS in the rat kidney tissue were detected by immunohistochemitry and morphometry computer image analysis.The histological changes of kidney were observed under light microscope.Results The expressions of HO-1 and iNOS were markedly higher,and the levels of SCr and BUN were also significantly higher in intestinal IRI model group than those in the sham operation group(all P
5.Application of sternocleidomastoid island myocutaneous flap in head and neck neoplasm's operation
Guohao WU ; Fujin CHEN ; Zhongyuan ZHENG
Chinese Journal of Microsurgery 1998;0(01):-
Objective To evaluate the effect of sternocleidomastoid island myocutaneous flap applicating in reconstructing defect of head and neck neoplasms after operation. Methods We used sternocleidomastoid island myocutaneous flap to restore defect postoperation of head and neck neoplasms for 9 patients. Five patients used flap to reconstruct defect of oral mucosal, one case pucker myocutaneous flap, part of it restore oral mucosal defect, part restore skin defect of cheek, the others reconstruct defect of cheek or parotid gland. Seven cases used polyhole titanium to reconstruct mandible bone at the same time. Results Eight cases used sternocleidomastoid island myocutaneous flap survival postoperation, one case's flap appeared necrosis of distant part and recover after one half month. Conclusion Pedical sternocleidomastoid island myocutaneous flap can provide huge flap supply, survival rate higher, easy to execute and not complex, it is suitable for clinical doctors to reconstruct defect after operation for head and neck tumors.
6.Effect of radix paeoniae rubra on expression of heme oxygenase and inducible nitric oxide synthesis in lipopolysaccharide-induced acute lung injury in rats
Zhongyuan XIA ; Chang CHEN ; Xiaoyuan WANG
Chinese Journal of Trauma 2003;0(09):-
Objective To investigate the effect of radix paeoniae rubra (RPR) on expression of heme oxygenase (HO-1) and inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) and explore its protective mechanism. Methods Forty Wistar rats were randomly and equally divided into five groups, ie, control group, LPS group, RPR treatment group, RPR prevention group and Hemin group. Arterial blood was drawn for blood gas analysis. Models of endotoxin-induced ALI were used to observe the protein content, the ratio of neutrophiles in bronchoalveolar lavage fluid (BALF), the malondialdehyde (MDA) content in lung and the activities of serum NO. Expression of HO-1 and iNOS in rat lung tissue was detected by immunohistochemistry and morphometry computer image analysis. The histological change of lung were observed under light microscope. Results Compared with control group, expression of HO-1 and iNOS was markedly increased (P
7.Suppression of intestinal epithelium apoptosis by shenfu solution during reperfusional period
Qingtao MENG ; Zhongyuan XIA ; Xianyi LIU ; Xiangdong CHEN ; Guixian XIONG
Chinese Journal of Tissue Engineering Research 2005;9(47):172-173
BACKGROUND: The abnormal apoptosis of intestinal epithelial cells is the main cause of intestinal mucous membrane injury during ischemia reperfusion. Shenfu injection has good therapeutic effect on intestinal mucous membrane injury. OBJECTIVE: To investigate the influence of shenfu injection medication on intestinal apoptotic epithelial number, apoptosis-related caspase-3 and Bcl-2 gene expression, as well as the level of tumor necrosis factor (TNF)in intestinal ischemia reperfusion rat model.DESIGN: Randomized controlled experiment.SETTING:Department of Anesthesia, Renmin Hospital of Wuhan University.MATERIALS:This experiment was carried out in the laboratory of the Department of Anesthesia, Renmin Hospital of Wuhan University, between December 2002 and June 2003. Totally 24 healthy SD rats of clean grade were randomized into blank control group, intestinal ischemia reperfusion (IR) group and shenfu injection group with 8 rats in each group.METHODS: Rats in each group were anesthetized with intraperitoneal injection of ethyl carbamate at dosage of 1mg/kg; then in intestinal IR group and shenfu injection group rats' superior mesenteric artery was occluded with vascular clamp for 1 hour before 2-hour reperfusion,which was not conducted in blank control group. Rats in shenfu injection group were intravenously injected with shenfu injection at 30 minutes before occlusion at dose of 0.02 mL/g, which was replaced by the same volume of physical saline in blank control group and intestinal IR group. The expression of caspase-3 and Bcl-2 protein and cell apoptosis wee detected.MAIN OUTCOME MEASURES: ① Comparison of apoptotic index between groups. ② The expression of caspase-3 and Bcl-2 gene in rat intestinal epithelium. ③ Comparison of TNF content in intestinal mucous membrane homogenate between groups.RESULTS: Totally 24 rats were included in this experiment and all ehtered the result analysis. ①Comparison of apoptotic index between groups:The apoptotic index was obviously lower in shenfu injection group than in intestinal IR group, but higher than in blank control group [(7.75-±1.89)%,(28.25±8.50)%, (4.25-±2.63)%, P < 0.01]. ② The expression of caspase-3 and Bcl-2 gene in rat intestinal epithelium: caspase-3 expression was lower in shenfu injection group than in intestinal IR group, but higher than in blank control group [(0.211 6±0.087 5), (0.354 7±0.077 8), (0.194 1±0.057 4) A,P < 0.01, P > 0.05]; Bcl-2 expression was remarkably higher in intestinal IR group than in blank control group (P < 0.05), but obviously reduced in shenfu injection group compared to intestinal IR group (P < 0.01). ③ TNF content of intestinal mucous membranehomogenate in each group: TNF content was remarkably higher in intestinal IR group than in blank control group and shenfu injection group [(189.7±56.3), (38.6±10.4), (47.5±l8.7)mg/L,P < 0.01], and basically the same in shenfu inj~tion group and blank control group (P > 0.05).CONCLUSION: Shenfu injection can suppress intestinal epithelial apoptosis by reducing TNF content and caspase-3 e.pression as well as upregulating the expression of Bcl-2 gene during ischemia reperfusion, thereby attenuating ischemia reperfusion injury of intestinal mucous membrane.
8.Protective effect of shenfu injection on intestinal mucosa during shock /resuscitation
Zhongyuan XIA ; Zhigang LIU ; Yuhong HE ; Xiangdong CHEN
Chinese Journal of Tissue Engineering Research 2005;9(26):257-259
BACKGROUND: Reperfusion injury and inadequate perfusion exist,causing the injury of intestinal mucosa during shock / resuscitation.OBJECTIVE: To investigate protective effect of shenfu injection (SFI) in equal dosage for human effect on intestinal mucosal pH (PHi) of sigmoid colon, the contents of nitric oxide (NO), malonialdehyde(MDA) and Ca2+ of intestinal mucosa, activity of serum diamine oxidase(DAO of rabbits during shock/resuscitation.DESIGN:A randomized controlled trial with experimental animals as the subjects.SETTING: DepartmentofAnesthesiology, Renmin Hospital of Wuhan University MATERIALS: The experiment was performed in the Research Institute of Anesthesiology, Renmin Hospital of Wuhan University from August to October in 2003. Twenty-four adult healthy white rabbits were randomly divided into 3 groups with 8 animals for each: Shenfu injection group (SFI group), shock/resuscitation group and control group .INTERVENTIONS: Hemorrhagic shock was induced by blood withdrawal from femoral artery at a rate of 2 mL/kg per minute until MAP dropped to 40 mmHg. MAP was maintained at this level for 60 minutes,then the collected auto-blood and the same volume of balanced salt solution was transfused to the animal for preparing shock/resuscitation model. In SFI group: the animals were given 2.1 mL/kg SFI together with the auto-blood transfusion and followed by a continuous infusion of 5mL/kg SFI per hour.MAIN OUTCOME MEASURES: Intestinal mucosal pH (PHi) of sigmoid colon, NO, MDA and Ca2+ contents of intestinal mucosa, activity of serum DAO were detected before shock (So), 1 hour after shock (S1), 1 hour (R1) and 3 hour (R3) following resuscitation.the R1 reperfusion (7.171±0.102) , and R3 reperfusion (7.194±0.106) of the SFI group were higher than those of the shock/ resuscitation group (6.920±0.155,6.971±0.165,P < 0.05,P < 0.01) and those of the contents of the reperfusion R1 (35.090±1.184) μkat/L and R3 (32.440±2.884) μkat/L of the SFI groups were significantly higher than those of the shock/resuscitation group [(50.994±2.684),(52.377±1.217) μkat/L,P< 0.01] and of the control group [(15.970±1.734), (16.620±0.767)μkat/L,Pgroup were significantly lower than those in the shock/resuscitation group [ (61.8±5.3,72.2±5.8 ) μmol/g , (68.2±4.9,96.9±8.5) μmol/L ,P< 0.05]. Ca2+ content of intestinal mucosa at reperfusion 3 hours in the SFI group [(2.43±0.27)μmol/L] was lower than that in the shock/resuscitation group [(2.93±0.34)μmol/L,P < 0.05] and higher than that in the control group [(2.26±0.31 )μmol/L, (P < 0.05)].CONCLUSION: SFI in equal dosage for human effect could protect intestinal mucosa from shock/resuscitation injury through improving perfusion and oxygenation, inhibit the activity of NO, reduce oxygen free radical and calcium overload, with a good protective effect on intestinal mucosa during shock/resuscitation.
9.Pretreatment with docosahexaenoic acid enhances the protective effect of angiopoietin-1 on rat brain microvascular endothelial cells during oxygen glucose deprivation
Xiaobo CHEN ; Zhongyuan XIA ; Fan LEI ; Rui XUE
Chinese Journal of Clinical Nutrition 2015;23(2):103-109
Objective To evaluate the role of pretreatment with docosahexaenoic acid (DHA) in the protective effect of angiopoietin-1 (Ang-1) on rat brain microvascular endothelial cells (BMVECs) during oxygen glucose deprivation (OGD).Methods BMVECs were sub-cultured in vitro and divided with a random number table into 7 groups:normal control group,normal control + Ang-1 group,OGD group,OGD + Ang-1 group,OGD + DHA group,OGD + DHA + Ang-1 group,and OGD + DHA + GW9662 + Ang-1 group.The normal control and normal control + Ang-1 groups were cultured in DMEM containing serum,5 mmol/L glucose,and 1.25 mmol/L pyruvate;OGD groups were cultured in glucose-and serum-free DMEM.DHA (40 μmol/L) was added to OGD + DHA,OGD + DHA + Ang-1,and OGD + DHA + GW9662 + Ang-1 groups,and 5 μmol/LGW9662 [inhibitor of peroxisome proliferator-activated receptor gamma (PPAR-γ)] to OGD + DHA + GW9662 +Ang-1 group,before pretreatment for 1 hour in 5% CO2 and 95% air.After the pretreatment,Ang-1 (250 ng/ml)was added to normal control + Ang-1,OGD + Ang-1,OGD + DHA + Ang-1,and OGD + DHA + GW9662 +Ang-1 groups.The cells were cultured in 94% N2∶ 5% CO2∶ 1% O2 for 24 hours,except for normal control and normal control + Ang-1 groups,which were cultured in 5% CO2 and 95% air instead.The cell apoptosis rate was detected with flow cytometry,expressions of Bax,Bcl-2,caspase-3,Tie-1,Tie-2,pTie-2,pAkt,ZO-1proteins with Western blot.Results Compared with normal control group,the cell apoptosis rate in OGD,OGD + Ang-1,OGD + DHA,OGD + DHA + Ang-1,and OGD + DHA + GW9662 + Ang-1 groups were significantly increased (P =0.000,0.000,0.000,0.004,0.000);the expression levels of Bax,caspase-3,and Tie-1 were significantly enhanced (Bax:0.62 ± 0.03,0.38 ± 0.03,0.45 ± 0.03,0.26 ± 0.02,0.33 ± 0.02vs.0.16 ±0.01;caspase-3:0.76 ±0.05,0.42 ±0.04,0.52 ±0.02,0.32 ±0.02,0.40 ±0.02 vs.0.15 ±0.01;Tie-1:0.51 ±0.03,0.25 ±0.01,0.33 ±0.02,0.16±0.01,0.22±0.02 vs.0.12 ±0.01;all P=0.000);the expression levels of Bcl-2,Bcl-2/Bax,Tie-2,and pTie-2 were significantly decreased [Bcl-2:0.09±0.01,0.20±0.01,0.16±0.02,0.31±0.01,0.22±0.01 vs.0.34±0.01;Bcl-2/Bax:(14.93±1.86)%,(68.03±5.56)%,(36.93 ±2.22)%,(119.1 ±13.3)%,(64.23 ±6.07)% vs.(208.33 ±7.37)%;Tie-2:0.07±0.01,0.16±0.02,0.11 ±0.01,0.21±0.01,0.18±0.01 vs.0.26±0.01;pTie-2:0.05±0.01,0.15 ±0.01,0.07 ±0.01,0.22 ±0.02,0.16±0.01 vs.0.27 ±0.01;allP=0.000],in addition,pAkt and ZO-1 in OGD,OGD + Ang-1,OGD + DHA,and OGD + DHA + GW9662 +Ang-1 groups were also significantly reduced (pAkt:0.13 ±0.01,0.26 ±0.01,0.14 ±0.01,0.28 ±0.02vs.0.39±0.02;ZO-1:0.08±0.01,0.18±0.01,0.10±0.01,0.19±0.01vs.0.23±0.02;allP=0.000).Compared with the OGD group,OGD + Ang-1 and OGD + DHA + Ang-1 groups demonstrated significantly reduced cell apoptosis (both P =0.000).Compared with normal control + Ang-1 group,the expression levels of pTie-2,pAkt,and ZO-1 were significantly decreased in OGD + Ang-1 and OGD + DHA +Ang-1 groups (pTie-2:0.15 ±0.01,0.22 ±0.02 vs.0.52 ±0.02;pAkt:0.26 ±0.01,0.37 ±0.04 vs.0.67 ± 0.05;ZO-1:0.18 ± 0.01,0.24 ± 0.02 vs.0.39 ± 0.05;all P =0.000).Compared with OGD +Ang-1 group,OGD + DHA + Ang-1 group had significantly decreased cell apoptosis and expression levels of Bax,caspase-3 and Tie-1,and significantly increased expressions of Bcl-2,Bcl-2/Bax,Tie-2,pTie-2,pAkt,and ZO-1 (all P =0.000),while OGD + DHA + GW9662 + Ang-1 group showed no significant differences in these indexes (P =0.202,0.770,0.382,0.448,0.233,0.736,0.143,0.526,0.495,0.670).Conclusion Pretreatment with DHA may enhance the protective effect of Ang-1 on rat BMVECs under the condition of OGD,possibly via activating PPAR-γ,suppressing Tie-1 expression,and hence amplifying the activation of Tie-2.
10.Blood glucose control improves myocardial glucose metabolism in diabetic rats
Xiaolin CHEN ; Zhongyuan WEN ; Teng WANG ; Yong WU
Basic & Clinical Medicine 2006;0(08):-
Objective To investigate the effects of glycemic control on myocardial glucose metabolism in type 2 diabetic rats. Methods Eighteen male Sprague-Dawley rats were divided into three groups:control, diabetes, therapy.Fed with high-fat diet and intraperitoneally injected with streptozotocin to establish type 2 diabetic model,the rate of carbohydrate oxidation was measured by isolated heart Langendorf perfusion apparatus, the expression level of glucose transport 4(GLUT4) was measured by Western blotting. Results Compared with the control group,the diabetic rats had a significantly depression of glucose uptake in the hearts(P