Effects of Nrf2-autophagy pathway in rats of the incisional pain-remifentanil-induced hyperalgesia / 天津医药

Objective To investigate the effect of nuclear factor erythroid 2-related factor 2(Nrf2)-autophagy pathway on the incisional pain-remifentanil-induced hyperalgesia of rat model. Methods Twenty-four male Sprague-Dawley rats were randomly allocated into three groups:saline+incisional pain group(group I),remifentanil+incisional pain group(group RI)and Nrf2 agonist t-BHQ group(group tBHQ),with 8 rats in each group.In group I and RI,normal saline at 0.1 mL/(kg· min) and remifentanilat 1 μg/(kg·min) were respectively infused into caudal vein for 60 min. Rats in group t-BHQ were injected intraperitoneally with Nrf2 agonist t-BHQ(15 mg/kg),the first time at 0.5 h before remifentanil infusion,per 12 h once,4 times in a row,the rest management was the same as group RI.Brennan incision pain model rats were constructed along with the infusions in the three groups. The thermal paw withdraw latency (PWL) and mechanical paw withdraw threshold(PWT)were measured at 24 h before the infusion(T0)and at 2 h(T1),6 h(T2),24 h(T3),48 h(T4)after the infusion. Rats were sacrificed after the tests, then the L4-6segments of signal cord were removed and the expression levels of autophagy-related proteins,microtubule associated protein 1 light chain 3Ⅱ(LC3Ⅱ),Beclin 1,Nrf2 and Nrf2 downstream molecular hemeoxygenase-1 (HO-1) were detected by Western blot assay. Results The PWT and PWL values were decreased significantly with the prolongation of the processing time in the three groups. Compared with group I, PWL and PWT values were decreased at T1-4,the expression levels of LC3Ⅱand Beclin-1 were increased while Nrf2 and HO-1 were decreased at T4in group RI (P<0.05). While compared with group RI, PWL and PWT values were increased, and the expressions of Nrf2 and HO-1 were increased, LC3Ⅱ and Beclin-1 protein were upregulated in group tBHQ (P<0.05).Conclusion The activation of Nrf2-autophagy pathway can improve the incisional pain-remifentanil induced hyperalgesia.

Effects of intrathecal injection of ginsenoside Rg1 on the level of glutamate transporter in the arthritis rats with chronic morphine tolerance / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effects of intrathecal injection of ginsenoside Rg1 at different doses on the changes of the behavior and the expressions of excitatory amino-acid transporter 1 (EAAT1), i. e., glutamate-aspartate transporter (GLAST) in the spinal dorsal horn of the arthritis rats with chronic morphine tolerance, and further to explore its mechanisms for morphine tolerance.</p><p><b>METHODS</b>After successful intrathecal injection, an adjuvant arthritis model was established in 36 healthy male SD rats. They were randomly divided into 6 groups, 6 in each group. They were intrathecally injected with 10 microL normal saline (Group NS), 10 microg morphine (Group M), 10 microg morphine + 50 microg ginsenoside Rg1 (Group MG50), 10 microg morphine +100 microg ginsenoside Rg1 (Group MG100), 10 microg morphine + 200 microg ginsenoside Rg1 (Group MG200), and 100 microg ginsenoside Rg1 (Group G100), respectively. The normal saline and morphine were intrathecally injected twice daily, while ginsenoside Rg1 at different doses was intrathecally injected once daily, for 7 successive days. Fifty percent mechanical paw withdrawal threshold (PWT) was dynamically detected to evaluate their behaviors. The rats were sacrificed on day 7 after medication. The L3-L5 segment of the spinal cord was isolated for determining the expression of GLAST in the spinal dorsal horn using immunofluorescence staining.</p><p><b>RESULTS</b>The PWT of Group M was significantly higher than that of Group NS on the 1st and 3rd day after medication (P < 0.05). But it was gradually shortened along with the increasing days of medication. There was no statistical difference between Group M and Group NS on the 7th day (P > 0.05), indicating the formation of morphine tolerance. The PWT of Group MG100 also showed a decreasing tendency, but obviously slower than that of Group M (P < 0.05). The PWT of Group G100 was higher than that of Group NS (P < 0.05). Compared with Group NS, the expression of GLAST in the spinal dorsal horn of rats in Group M was down-regulated (P < 0.01). Compared with Group M, the expression of GLAST in the spinal dorsal horn of rats in Group MG100 and Group G100 was up-regulated (P < 0.05).</p><p><b>CONCLUSIONS</b>Single application of ginsenoside Rg1 showed mild antinociceptive effect in adjuvant-induced arthritis rats. Intrathecal injection of 100 microg ginsenoside Rg1 could attenuate the formation of morphine tolerance. Its mechanisms might be correlated with up-regulating of the expression of GLAST.</p>

Monocyte response and regulatory effect of emodin and shenmai injection on it in patients with severe sepsis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the impact of monocyte releasing tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) stimulated by lipopolysaccharide (LPS) in severe sepsis (SS) patients, and the regulatory effect of emodin and Shenmai Injection (SMI) on monocyte response.</p><p><b>METHODS</b>ocyte (PBMC) sampled from SS patients due to severe abdominal inflammation and healthy controls, PBMC were incubated with or without LPS, respectively. PBMC media pretreated with emodin and SMI, and then the levels of cytokine factors including TNF-alpha and IL-10 in supernatants were determined after stimulated with or without LPS in the two groups. Normal PBMC stimulated with LPS based on incubated with either normal serum or SS serum, and the levels of TNF-alpha and IL-10 in supernatant of normal PBMC and SS serum dealing with emodin or SMI after cultured and stimulated with LPS were determined, respectively.</p><p><b>RESULTS</b>The levels of TNF-alpha and IL-10 were significantly higher in SS patients than those in the healthy adults. The contents of TNF-alpha and IL-10 increased in response to LPS in PBMC of healthy adults, and the excretion of TNF-alpha was significantly attenuated whereas IL-10 significantly increased in septic PBMC than basal content. Both of the two traditional Chinese medicines had significantly effect in stimulating PBMC secretion in healthy adults and SS patients. In normal PBMC, emodin attenuated TNF-alpha and IL-10 release in response to LPS, and SMI significantly inhibited TNF-alpha release. As to septic PBMC, emodin significantly stimulated TNF-alpha and IL-10 release and SMI significantly increased the concentration of TNF-alpha in the SS patients. Incubation of normal PBMC with septic serum attenuated TNF-alpha production, whereas increased IL-10 release. Emodin could significantly decrease the level of IL-10, and SMI recovered TNF-alphareleasing and had no effect on IL-10.</p><p><b>CONCLUSION</b>The level of TNF-alpha significantly decreased accompanied with an anti-inflammatory cytokine IL-10 significantly increased of PBMC in SS patients. Monocyte exhibits the different response of inflammatory or anti-inflammatory factor which may be one of the reasons of imbalance of immune function in SS patients. Both of emodin and SMI may have regulatory effect on excretion of PBMC in SS patients, but they play a role in different ways.</p>

Change of TH1/TH2 cytokine equilibrium in rats with severe sepsis and therapeutic effect of recombinant interleukin-12 and Shenmai injection / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the dynamic change of Th1/Th2 cytokines in serum, peritoneal lavage fluid (PLF) and splenic macrophages (SM) in rats with severe peritonitis, and to observe the therapeutic effects of recombinant interleukin-12 (rIL-2) and Shenmai injection (SMI), a Chinese medicinal preparation.</p><p><b>METHODS</b>Severe peritonitis (SP) model was induced by intraperitoneal injection of E. coli and B. frag, and mild peritonitis (MP) model was induced by cecal ligation and punching. Then the following experiments were done: (1) Survival rates of animals after every 6 hrs in the 72 hrs after modeling were recorded, serum and PLF levels of cytokines, including tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), and interleukin-10 (IL-10), 6 hrs, 12 hrs, 24 hrs and 48 hrs after modeling were measured. (2) Model rats were treated with rIL-12 or SMI, the survival rate was recorded and serum levels of TNF-alpha, INF-gamma, and IL-10 before and after treatment were measured, and (3) amount of these cytokines produced by SM were determined 6 hrs, 12 hrs and 24 hrs after treatment. The survival rates and levels of cytokines were then compared between the groups (model group treated with rIL-12 or SMI, untreated model group, and blank group).</p><p><b>RESULTS</b>Serum and PLF levels of IFN-gamma, TNF-alpha at all the time points in SP rats were significantly lower than those in MP rats while those of IL-10 6 hrs and 12 hrs after modeling were significantly higher in the former than that in the latter (P < 0.05). IFN-gamma secretion of SM in SP rats was significantly higher than that in MP rats 6 hrs after modeling (P < 0.05). Administration of rlL-12 or SMI given before modeling could improve the survival rate of the model rats (P < 0.05) and cause significant increase of the serum level and SM secretion of IFN-gamma.</p><p><b>CONCLUSION</b>Imbalance in promoting/antagonizing inflammatory cytokines and Th2 response dominance appear in SP rats early at the initiating stage, and SM secretion of inflammation promoting factor also reduces. Administration in time of rIL-12 and SMI, may increase the survival rate, and its mechanism may be related with their immuno-stimulating action.</p>

Anesthetic efficacy of epidural ropivacaine mixed with chloroprocaine for hysterectomy / 中华麻醉学杂志

Objective To assess the efficacy of the mixtures of different concentrations of ropivacaine(R) and chloroprocaine(C)for epidural anesthesia in patients undergoing hysterectomy.Methods Sixty ASAⅠorⅡpatients aged 27-56 weighing 45-75 kg undergoing elective hysterectomy were randomly divided into 4 groups(n= 15 each);groupⅠ0.75% R alone;groupⅡ0.5% R-1% C;groupⅢ0.5% R-1.5% C and groupⅣ0.75% R+1% C.The epidural catheter was placed at L_(2,3)interspace and advanced 3.5 cm into the epidural space in cephalad direction.A total of 22 ml of epidural solution was injected in each group.The onset time,block height and duration of sensory block and the onset time,degree and duration of motor block(using Bromage scale)were assessed.The use of supplementary drugs(ketamine and ephedrine)and side effects were recorded.Results The onset time of sensory and motor block was significantly shorter in groupⅡ,ⅢandⅣthan in groupⅠ(0.75% R alone).The duration of sensory and motor block was significantly shorter in groupⅡandⅢthan in groupⅠandⅣ.The incidence of hypotension was significantly increased but the incidence of discomfort produced by traction on the viscera during operation was reduced in groupⅣas compared with groupⅠ.Conclusion The anesthetic efficacy of epidural 0.5% ropivacaine is significantly enhanced when mixed with 1.0% or 1.5% chloroprocaine.