ABSTRACT
Objective To investigate the toxicity of lidocaine solid lipid nanoparticles (SLNs) in human neurons.Methods Lidocaine-loaded SLNs were prepared using high pressure homogenization.SHSY5Y cells were cultured in vitro and inoculated on 96-well plates (100 μl/well) at a density of 5× 105 cells/ml.SH-SY5Y cells were randomized into 10 groups (n =30 each) using a random number table:control group (group C), different concentrations of lidocaine groups (L1-4 groups), different concentrations of lidocaine SLN groups (L-SLN1-4 groups), and blank SLN group (group SLN).The cells were cultured routinely in group C.The cells were incubated with the culture medium containing lidocaine with the final concentrations of 1.000％, 0.500％, 0.250％ and 0.125％ in L1-4 groups, respectively.In LSLN1-4 groups, the cells were incubated with the culture medium containing lidocaine SLNs with the final concentrations of 1.000％, 0.500％, 0.250％ and 0.125％ in L1-4 groups, respectively.Before incubation (at the corresponding time points in group C), and at 1, 12 and 24 h of culture or incubation (T0-3) , 6 wells in each group were selected for measurement of the cell survival rate (using methyl thiazolyl tetrazolium assay).The cell morphology was examined with optical microscope at T3.Results Compared with that at T0, the cell survival rate was significantly decreased at each time point in L1-4 and L-SLN1,2 groups, at T2,3 in L-SLN3 group, and at T3 in L-SLN4 group (P＜0.05) , and no significant change was found in SLN and C groups (P＞0.05).The cell survival rate was significantly lower at T2,3 in L1-4 and L-SLN1-3 groups, and at T3 in group L-SLN4 than that at T1, and at T3 in L1-4 and L-SLN1-4 groups than that at T2 (P＜0.05).Compared with group C, the cell survival rate was significantly decreased at each time point in L1-4 and L-SLN1,2 groups, at T2,3 in group L-SLN3, and at T3 in group L-SLN4 (P＜0.05) , and no significant change was found in group SLN (P＞0.05).Compared with group L-SLN at the corresponding concentration, the cell survival rate was significantly decreased at each time point in group L1-4 (P＜0.05).Conclusion Lidocaine SLNs have toxic effect on human neurons, but the effect is weaker than that caused by Iidocaine solution.
ABSTRACT
Objective To investigate the effect of electro-acupunctare at zusanli on acute lung injury in a rat model of sepsis after scald.Methods Fifty SPF male Sprague-Dawley rats,weighing 200-250 g,aged 2-3 months,were randomly divided into 5 groups (n =10 each) using a random number table:control group (group C),sepsis after scald group (group SS),electro-acupuncture at zusanli group (group E),electric stimulation of non-acupoint group (group NE) and electro-acupuncture at zusanli + α-bungarotoxin (α-BGT,a selective α7 nicotinic acetylcholine receptor antagonist) group (group α-BGT).The rats were subjected to a third degree scald covering 20％ total body surface (TBS) and muramyl dipeptide (MDP) 5 mg/kg was injected into the femoral vein immediately after scald to induce sepsis.Electro-stimulation (3 V,2 ms,3 Hz) of bilateral zusanli was performed for 12 min starting from the time point immediately after MDP injection and every 8 h for 2 consecutive days in group E.In group NE,electro-stimulation was performed at the points 5 mm lateral to the bilateral acupoints of Zusanli and the method was similar to those previously described in group E.In group α-BGT,α-BGT 1.0 μg/kg (in 1 ml of normal saline) was injected into the femoral vein before electro-stimulation of zusanli.At 48 h after treatment,arterial blood samples were obtained for determination of serum tumor necrosis factor-alpha (TNF-α) and high mobility group box-1 (HMGB1) protein levels (by ELISA) and lung specimens were removed for microscopic examination and for determination of the expression of Nod like receptor 2 (NLR2) mRNA (by RT-PCR) and receptor interacting protein 2 (RIP2) in the lung tissues (by Western blot).Results Compared with group C,the expression of NLR2 mRNA and RIP2 was significantly up-regulated,and the serum TNF-α and HMGB1 levels were increased in SS,NE and α-BGT groups (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group E (P ＞ 0.05).Compared with group SS,the expression of NLR2 mRNA and RIP2 was down-regulated,and the serum TNF-α and HMGB1 levels were decreased in group E (P ＜ 0.05),and no significant change was found in the parameters mentioned above in NE and α-BGT groups (P ＞ 0.05).Compared with group E,the expression of NLR2 mRNA and RIP2 was significantly up-regulated,and the serum TNF-α and HMGB1 levels were increased in NE and α-BGT groups (P ＜ 0.05).The pathological changes of lung tissues were significantly reduced in group E as compared with group SS.Conclusion Electro-acupunctare at Zusanli can reduce acute lung injury in a rat model of sepsis after scald and inhibition of NLR2/RIP2 signaling pathway and activation of cholinergic anti-inflammatory pathway in lung tissues may be involved in the mechanism.
ABSTRACT
Objective To evaluate the effect of electro-acupunctare at Zusanli on liver injury in a rat model of sepsis after scald.Methods Fifty SPF male Sprague-Dawley rats,weighing 200-250 g,aged 2-3 months,were randomly divided into 5 groups (n =10 each) using a random number table:control group (group C); sepsis after scald group (group SS); electro-acupuncture at Zusanli group (group E); electric stimulation of non-acupoint group (group NE); electro-acupuncture at Zusanli + α-bungarotoxin (α-BGT) group (group α-BGT).The rats were subjected to a third degree scald covering 20％ of the total body surface area (TBSA) and sepsis was induced with muramyl dipeptide (MDP) 5 mg/kg injected into the femoral vein immediately after scald.Electro-stimulation (3 V,2 ms,3 Hz) of bilateral Zusanli acupoints or non-acupoints was performed for 12 min every 8 h for 2 consecutive days starting from the time point immediately after MDP was injected in group E.In group NE,electro-stimulation was performed at the points 5 mm lateral to the acupoints of bilateral Zusanli and the method was similar to those previously described in group E.In group α-BGT,α-BGT 1.0 μg/kg (in normal saline 1 ml) was injected into the femoral vein before electric stimulation,then electro-stimulation was performed and the method was similar to those previously described in group E.After 48 h of continuous stimulation,liver specimens were obtained for microscopic examination of the pathological changes.The blood samples were obtained from the abdominal aorta for determination of the levels of serum ALT and AST,tumor necrosis factor-alpha (TNF-α),and high mobility group box-1 (HMGB1) (by ELISA),and expression of Nod-like receptor 2 (NLR2) mRNA (by RTPCR) and receptor-interacting protein 2 (RIP2) in the liver tissues (by Western blot).Results The serum ALT and AST,TNF-α and HMGB-1 levels and NLR2 mRNA and RIP2 expression in liver tissues were significantly higher in SS group than in C group.Compared with SS group,the serum ALT,AST,TNF-α and HMGB1 levels were significantly decreased and NLR2 mRNA and RIP2 expression was down-regulated in E group,and no significant changes were found in the parameters mentioned above in NE group.Compared with E group,the serum ALT,AST,TNF-α and HMGB1 levels were significantly increased,and NLR2 mRNA and RIP2 expression was up-regulated in α-BGT group.The pathological changes in liver tissues were significantly reduced in group EA as compared with SS,NE and α-BGT groups.Conclusion Electro-acupuncture at Zusanli can reduce liver injury in a rat model of sepsis after scald and inhibition of NLR2/RIP2 signaling pathway and activation of cholinergic antiinflammatory pathway mediated by α7 nicotinic acetylcholine receptors in liver tissues may be involved in the mechanism.