Effect of moxibustion on immune function homeostasis in rats with diarrhea irritable bowel syndrome based on SCF/c-kit signaling pathway / 中国针灸

OBJECTIVE@#To observe the effects of moxibustion on the stem cell factor (SCF)/tyrosine kinase receptor (c-kit) signaling pathway and immune function in rats with diarrhea irritable bowel syndrome (IBS-D), and to explore the mechanism of moxibustion for IBS-D.@*METHODS@#Among 52 young rats born from 6 healthy pregnant SPF rats, 12 rats were randomly selected into the normal group, and the remaining 40 rats were treated with the three-factor combination method of maternal separation, acetic acid enema and chronic restraint stress to establish the IBS-D rat model. Thirty-six rats with successful IBS-D model were randomly divided into a model group, a moxibustion group, and a medication group, 12 rats in each group. The rats in the moxibustion group were treated with suspension moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37); the rats in the medication group were treated with intragastric administration of rifaximin suspension (150 mg/kg). All the treatments were given once a day for 7 consecutive days. The body mass, loose stool rate (LSR), the minimum volume threshold when abdominal withdrawal reflex (AWR) scored 3 were measured before acetic acid enema (35 days old), after modeling (45 days old), and after intervention (53 days old). After intervention (53 days old), HE staining was used to observe the morphology of colon tissue, and spleen and thymus coefficients were measured; ELISA method was used to detect serum inflammatory factors (tumor necrosis factor a [TNF-a], interleukin [IL]-10, IL-8), T-lymphocyte subsets (CD+4, CD+8, CD+45), value of CD+4/CD+8 and immune globulin (IgA, IgG, IgM); real-time PCR method and Western blot method was used to detect the expression of SCF, c-kit mRNA and protein in colon tissue; immunofluorescence staining method were used to detect positive expression of SCF and c-kit.@*RESULTS@#After intervention, compared with the normal group, in the model group, the body mass and the minimum volume threshold when AWR scored 3 were decreased (P<0.01), LSR, spleen and thymus coefficients, serum levels of TNF-α, IL-8, CD+4, CD+45, CD+4/CD+8, IgA, IgG, IgM were increased (P<0.01), serum IL-10 level and protein and mRNA expression of SCF and c-kit in colon tissue were decreased (P<0.01), and the positive expression of SCF and c-kit was decreased (P<0.01). Compared with the model group, in the moxibustion group and the medication group, the body mass and the minimum volume threshold when AWR scored 3 were increased (P<0.01, P<0.05), LSR, spleen and thymus coefficients, serum levels of TNF-α, IL-8, CD+4, CD+8, CD+45, CD+4/CD+8, IgA, IgG, IgM were decreased (P<0.01, P<0.05), serum IL-10 level and protein and mRNA expression of SCF and c-kit in colon tissue were increased (P<0.01), and the positive expression of SCF and c-kit was increased (P<0.01). Compared with the medication group, in the moxibustion group, the level of serum CD+4 was decreased (P<0.05), the value of CD+4/CD+8 was increased (P<0.01), and there was no significant difference in other indexes (P>0.05). The expression of SCF and c-kit mRNA was positively correlated with the minimum volume threshold when AWR scored 3 and IL-10 (P<0.01), and negatively correlated with remaining indexes (P<0.01, P<0.05).@*CONCLUSION@#Moxibustion could reduce visceral hypersensitivity, improve symptoms of abdominal pain and diarrhea in IBS-D rats, and its mechanism may be related to up-regulation of the expression of SCF/c-kit signaling pathway and improvement of IBS-D immune function.

Anti-inflammation effect of moxibustion for rats with diarrhea-predominant irritable bowel syndrome based on multiple miRNAs regulating NF-κB signal pathway / 中国针灸

OBJECTIVE@#To observe the effect of moxibustion on the regulation of nuclear factor-kappa B (NF-κB) and inflammatory factors by multiple microRNAs (miRNAs) in rats with diarrhea-predominant irritable bowel syndrome (IBS-D), and to explore the anti-inflammatory mechanism of moxibustion on IBS-D.@*METHODS@#Twelve of 52 newborn rats were randomly selected into a normal group. The remaining rats were made into IBS-D model. A total of 36 rats with successful model were randomly divided into a model group, a medication group and a moxibustion group, 12 rats in each group. The rats in the medication group were intraperitoneally injected with pyrrolidine dithiocarbamate (PDTC). The rats in the moxibustion group were treated with moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) for 20 min each time. All the intervention was given once a day for 7 days. Before and after modeling as well as after intervention, the body mass, loose stool rate and the minimum volume threshold of abdominal withdrawal reflex (AWR) were measured. After intervention, the contents of serum tumor necrosis factor α (TNF-α), interleukin (IL)-1β and IL-8 were detected by ELISA method; the morphology of colon tissues was observed by HE staining, and the expressions of miR-155, miR-125b, miR-29b, miR-31, miR-18a and NF-κB p65 mRNA in colon tissues were detected by real-time PCR. The expressions of NF-κB p65, TNF-α, IL-1β and IL-8 protein in colon tissues were detected by immunofluorescence.@*RESULTS@#After modeling, the body mass and the minimum volume threshold of AWR in the model group were lower than those in the normal group (P<0.01); the rates of loose stool in the model group were higher than those in the normal group (P<0.01); after intervention, in the model group, the inflammatory infiltration of colon tissues was obvious, and the serum levels of TNF-α, IL-1 β, IL-8 were higher than those in the normal group (P<0.05); the expression of miR-155, miR-125b, miR-29b, miR-31, miR-18a and NF-κB p65 mRNA in colon tissues was higher than that in the normal group (P<0.05); the protein expression of NF-κB p65, TNF-α, IL-1β, IL-8 was also higher than that in the normal group (P<0.01). After intervention, the body mass and the minimum volume threshold of AWR in the medication group and the moxibustion group were both higher than those in the model group (P<0.05); the loose stool rate in the medication group and the moxibustion group were lower than those in model group (P<0.05); the inflammatory cells infiltration in the colon tissues was less, the serum levels of TNF-α, IL-1β and IL-8 as well as the protein expression of NF-κB p65, TNF-α, IL-1β and IL-8 in the colon tissues in the medication group and the moxibustion group were lower than those in the model group (P<0.05, P<0.01). The expression of miR-125b, miR-31, miR-18a and NF-κB p65 mRNA in the medication group were lower than those in the model group (P<0.05). The expression of miR-155, miR-125b, miR-29b, miR-31, miR-18a and NF-κB p65 mRNA in the moxibustion group were lower than those in the model group (P<0.05). The miR-155, miR-125b, miR-29b, miR-31, miR-18a were positively correlated with NF-κB p65 mRNA (0<r<1, P<0.01).@*CONCLUSION@#The anti-inflammatory mechanism of moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) for IBS-D rats may be related to regulating multiple miRNAs to inhibit NF-κB signal pathway and reduce the expression of inflammatory factors.

Effect of warm acupuncture at Yifeng (TE 17) on facial paralysis with periauricular pain during pregnancy / 中国针灸

OBJECTIVE@#To compare the curative effect between the warm acupuncture at Yifeng (TE 17) combined with conventional acupuncture and TDP plus conventional acupuncture on facial paralysis with periauricular pain during pregnancy.@*METHODS@#A total of 68 patients were randomized into an observation group (36 cases, 3 cases dropped off) and a control group (32 cases, 1 case dropped off). First week, TDP light was used on the affected side in the control group, and warm acupuncture at Yinfeng (TE 17) on the affected side was used in the observation group, both once a day. From the second week, both groups were given acupuncture at Chengjiang (CV 24) and the affected side of Cuanzhu (BL 2), Yangbai (GB 14), Taiyang (EX-HN 5), Yingxiang (LI 20), Dicang (ST 4), etc. and electroacupuncture (continuous wave, 2 Hz in frequency) was connected at Cuanzhu (BL 2) and Taiyang (EX-HN 5), Jiache (ST 6) and Dicang (ST 4). Both treatments were given every other day for 4 weeks totally. The visual analogue scale (VAS) score of the periauricular pain degree before treatment and after 1 week of treatment, the House-Brackmann (H-B) facial nerve function grading scale and facial disability index (FDI) score before treatment and after 2, 4 weeks of treatment were compared between the two groups.@*RESULTS@#After 1 week of treatment, the VAS scores of both groups decreased (@*CONCLUSION@#Warm acupuncture at Yinfeng (TE 17) combined with conventional acupuncture can effectively improve the periauricular pain and facial nerve function in patients of facial paralysis with periauricular pain during pregnancy, and the curative effect is better than TDP plus conventional acupuncture.

Clinical observation on time-effect of electroacupuncture for idiopathic facial paralysis / 中国针灸

OBJECTIVE@#To observe the effect of different acupuncture frequency and duration of needle retention on idiopathic facial paralysis, and optimize the acupuncture treatment plan.@*METHODS@#A total of 140 patients were randomized into a group A (37 cases, 3 cases dropped off), a group B (35 cases, 3 cases dropped off), a group C (34 cases, 1 case dropped off) and a group D (34 cases, 1 case dropped off). Under the same basic treatment, acupuncture intervention time (day 8 of morbidity), acupoint selection [Cuanzhu (BL 2), Yangbai (GB 14), Taiyang (EX-HN 5), Yingxiang (LI 20), Jiache (ST 6), Dicang (ST 4) on the affected side, Chengjiang (CV 24) and Hegu (LI 4) on the unaffected side] and electroacupuncture intervention, different acupuncture interval time and duration of needle retention were applied. In the group A, the treatment was given 20 min once a day, while the group B 30 min once a day, the group C 20 min once every 2 days, the group D 30 min once every 2 days. Totally 20-day treatment were required. The House-Brackmann (H-B) facial nerve function grading system was used to evaluate the improvement of clinical symptom, the situation and latency periods of the R1 wave in blink reflex and facial nerve motor conduction before and after treatment were observed in the 4 groups.@*RESULTS@#After treatment, the cured rates in the 4 groups were 44.1% (15/34), 46.9% (15/32), 57.6% (19/33) and 51.5% (17/33), there was no significant difference among 4 groups (>0.05). The situation and latency periods of the R1 wave in blink reflex and latency periods and amplitude of facial nerve motor conduction after treatment were improved in the 4 groups (0.05).@*CONCLUSION@#Acupuncture 20 min once a day, 30 min once a day, 20 min once every 2 days and 30 min once every 2 days have significant effect on the recovery of idiopathic facial paralysis, and the effect is comparable.

The effect of propofol preconditioning on cytochrome C release from mitochondria after mild hypothermic ischemia/reperfusion in isolated rat hearts / 中国应用生理学杂志

<p><b>AIM</b>To explore the effect of propofol preconditioning on cardiomyocyte apoptosis and cytochrome C release from mitochondria during mild hypothermic ischemia/reperfusion in isolated rat hearts.</p><p><b>METHODS</b>50 isolated SD rat hearts perfused on Langendorff apparatus were randomly divided into 5 groups (n=10): control group (C), DMSO group (D), 3 different concentrations of propofol groups of 25 micromol x L(-1) (P1), 50 micromol x L(-1) (P2), 100 micromol x L(-1) (P3) propofol respectively. All of the isolated rat hearts were subjected to 31 degrees C mild hypothermic ischemia for 55 min followed by 60 min reperfusion. The D, P1, P2, P3 groups were preconditioned by perfusing with K-H solution containing 20 micromol x L(-1) DMSO and 25, 50, 100 micromol x L(-1) propofol respectively for 10 min and then followed by 5 min K-H solution washing out before ischemia. The preconditioning procedure was repeated twice. Hemodynamics of the hearts was recorded after equilibration(baseline values) immediately before ischemia, 30 min and 60 min after reperfusion respectively. Cardiomyocyte apoptosis rate and contents of cytosolic and mitochondrial cytochrome C were measured at the end of reperfusion.</p><p><b>RESULTS</b>After 30 min and 60 min reperfusion, LVEDP was significantly lower and LVDP was significantly higher in P3 group than those in C group ( P < 0.05, P < 0.01). Compared with C group, cardiomyocyte apoptosis rate of the hearts decreased significantly in P2,P3 groups at the end of reperfusion (P < 0.05, P < 0.01). Cytochrome C level increased significantly in mitochondria but decreased significantly in cytosol in P2, P3 groups as compared with C group (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>Propofol preconditioning decreased cardiomyocyte apoptosis, protected the heart against 31 degrees C mild hypothermic ischemia/reperfusion injury by attenuation of the release of cytochrome C from mitochondria to cytosol.</p>

Protective effects of propofol preconditioning on myocardium against hypothermia ischemia normothermia reperfusion injury on isolated rat hearts / 上海第二医科大学学报

Objective To investigate the protective effects of propofol preconditioning on myocardium against hypothermia ischemia normothermia reperfusion injury on isolated rat hearts. Methods The Langendorff apparatus was used.Sixty SD rat hearts were divided randomly into 5 groups after 20-minute equilibrium(n=12): control(Con) group,hearts were continually perfused with K-H buffer for 175 min;ischemia/reperfusion(I/R) group,hearts were perfused with K-H buffer for 40 min,then subjected to global ischemia at 27 ℃ for 75 min,and followed reperfusion at 37 ℃ for 60 min;propofol preconditioning group 1(P1),group 2(P2),and group 3(P3),hearts were perfused with K-H buffer including 50,100,and 150 ?mol/L propofol for 10 min and followed reperfusion like I/R group,respectively.Heart rate(HR),left ventricular end-diastolic pressure(LVEDP), left ventricular developed pressure(LVDP) and ?dp/dtmax at the end of equilibration,pre-ischemia and at the end of reperfusion were recorded.The contents of creatine kinase(CK) and lactate dehydrogenase(LDH) in coronary effluent were measured at the end of equilibration and 1,10,20,30,and 60 min during reperfusion.The activity of superoxide dismutase(SOD) and the contents of maleic dialdehyde(MDA) were measured at the end of reperfusion.The area of infarct region was determined at the end of reperfusion. Results HR,LVDP,?dp/dtmax and SOD activity in P2 and P3 group were higher than those in I/R group(P

Inhibition of thermal hyperalgesia and tactile allodynia by intrathecal administration of gamma-aminobutyric acid transporter-1 inhibitor NO-711 in rats with chronic constriction injury / 生理学报

The present study was undertaken to explore the role of gamma-aminobutyric acid transporters in the neuropathic pain. On the chronic constriction injury (CCI) rats 4 doses (5, 10, 20, 40 microg in group N5, N10, N20, N40, respectively) of specific gamma-aminobutyric acid transporter-1 inhibitor NO-711 or normal saline (in group NS) were intrathecally administered before sciatic nerve ligation (pre-treatment) or at the third day after ligation (post-treatment). The paw withdrawl latency (PWL) from a noxious thermal stimulus and paw withdrawl mechanical threshold (PWMT) of von Frey filament was used as measure of thermal hyperalgesia and tactile allodynia respectively. The results demonstrated that post-treatment of NO-711 significantly suppressed thermal hyperalgesia and allodynia in CCI rats (P<0.05, P<0.01), the inhibitory effect lasted for 2 h (N40 group) and 4 h (N20 group) respectively. NO-711 inhibited thermal hyperalgesia induced by CCI in a dose-dependent manner. Intrathecal pretreatment with different doses of NO-711 delayed the occurrence of thermal hyperalgesia, but could not delay the emergence of allodynia induced by CCI. This study indicates that gamma-aminobutyric acid transporter inhibitor has anti-thermal hyperalgesia and anti-tactile allodynia effects in neuropathic rats.

Myocardioprotective effects of the combination of ischemic preconditioning with hypothermia and crystalloid cardioplegia in immature rabbits / 生理学报

This study was undertaken to explore the myocardioprotective effects of the combination of ischemic preconditioning (IP) with hypothermia and St.II Thomas crystalloid cardioplegic solution (CCS) on immature hearts in the rabbit. Isolated immature rabbit hearts were perfused with Krebs-Henseleit bicarbonate buffer on Langendorff apparatus. In experiment 1, 24 hearts were divided into 4 groups (n=6 in each group): Con, IP1, IP2 and IP3 group. Hearts of the four groups underwent 0, 1, 2 or 3 cycles of IP respectively. Then all the hearts were subjected to a sustained ischemia period of 2 h at 20 degrees C and a postischemic reperfusion period of 30 min at 37 degrees C. In experiment 2, 48 hearts were divided into 6 groups (n=8 in each group): SCon1, SIP1, SCon2, SIP2, SCon3 and SIP3 group, according to hypothermia and the duration of sustained ischemia (30 min at 32 degrees C; 90 min at 25 degrees C, 2 h at 20 degrees C). The SIP1, SIP2 and SIP3 groups were preconditioned twice before the sustained hypothermic ischemia, while the SCon1, SCon2 and SCon3 groups were not preconditioned. CCS was applied during sustained ischemia, all the hearts were reperfused for 30 min at 37 degrees C. Heart rate (HR), left ventricular developed pressure (LVDP) and peak rate of increase or decrease of left ventricular pressure (+/-dp/dt(max)) were recorded. Tissue concentration of adenosine triphosphate (ATP), malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were measured. At the end of reperfusion, values of product of LVDP and HR, +/-dp/dt(max) in IP2 group were 96%+/-21%, 101%+/-19% and 84% +/-15% of the baseline values respectively, which were significantly higher than those of Con group and IP3 group (P<0.01, P<0.05); also, the ATP content of IP2 group was higher than that of the Con group (P<0.01). When CCS was applied during sustained period of hypothermic ischemia at 32 degrees C or 25 degrees C, recovery rates of RPP (rate product, =LVDPxHR) and +dp/dt(max) in SIP1 group were 87% +/-14% or 99% +/-26% of the baseline values respectively (P<0.05, vs SCon1 group), the values in SIP2 group changed to 87% +/-16% or 102% +/-20% respectively (P<0.05, vs SCon2 group). Contents of ATP in SIP1 and SIP2 groups were significantly higher than those of SCon1 or SCon2 groups respectively (P<0.05), but MDA content of the two groups were significantly lower than those of SCon1 or SCon2 groups (P<0.05) respectively. The study indicates that IP attenuates hypothermic ischemia/reperfusion injury to immature rabbit hearts under 20 degrees C ischemia, two cycles of IP showing better myocardioprotective effects than 1 or 3 cycles of IP. When IP was combined with CCS which were applied during hypothermic ischemia period, the beneficial effects of IP were weakened as the temperature during the hypothermic period was elevated.

Effects of ischemic preconditioning on the hypothermic ischemia/reperfusion injury of immature rabbit hearts / 中国应用生理学杂志

<p><b>AIM</b>To study effects of ischemic preconditioning on the hypothermic ischemia/reperfusion injury of immature rabbit hearts.</p><p><b>METHODS</b>The isolated immature rabbit (3-4 weeks) hearts were perfused on Langendorff apparatus. After 30 min perfusing with 37 degrees C K-H perfusate, the hearts in part one were yielded 0, 1, 2 or 3 times of IP respectively before 120 min ischemia at 20 degrees C hypothermia, and the hearts in part two were yielded 0 or 2 times of IP before being arrested by infusion of St. Thomas II crystalloid cardioplegic solution, then the arrested hearts were yielded ischemia for 30, 90 or 120 min at 32 degrees C, 25 degrees C and 2 degrees C hypothermia respectively. Then all the hearts were reperfused for 30 min at 37 degrees C normothermia. Heart rate (HR), left ventricular developed pressure (LVDP), +/- dp/dt(max) were recorded at baseline, preischemic and 1, 3, 5, 10, 20, 30 min after reperfusion. Also contents of ATP and MDA and activity of SOD and Ca(2+) -ATPase of myocardium were measured.</p><p><b>RESULTS</b>At the end of reperfusion, the recovery rate of left ventricular function in IP2 group were significantly higher than that of control group and IP3 group (P < 0.01, P < 0.05), also the IP2 group showed a higher content of ATP and activity of Ca(2+) -ATPase than control group and IP3 group (P < 0.01, P < 0.05). When the ischemic hearts were at different hypothermia accompanied with CCS, the recovery rate of left ventricular function and contents of ATP in SIP1 and SIP2 group were significantly higher than that of SCon 1 group and SCon 2 group respectively (P < 0.01, P < 0.05), the contents of MDA in the two IP groups were lower than that of the two control groups.</p><p><b>CONCLUSION</b>IP can attenuate the hypothermic ischemia/reperfusion injury of immature rabbit hearts, the cardioprotective effects are dependent on the mode of IP and the possible mechanisms may involve the following aspects: decrease the consumption of ATP, inhibit lipid peroxidation and maintain the activity of Ca(2+) -ATPase of cardiac myocyte.</p>

Protective effect of isoflurane preconditioning on myocardium against ischemia-reperfusion injury in isolated rat hearts / 中华麻醉学杂志

Objective To investigate the protective effect of isoflurane preconditioning on myocardium against isehemia-reperfusion(I/R)injury and the possible mechanism.Methods Fifty male SD rats weighing 250- 300 g were randomly divided into 5 groups(n=10 each):Ⅰ control group(C);Ⅱ I/R group and 3 isoflurane preconditioning groups with 0.5%(Ⅲ),or 1.0%(Ⅳ)or 2.0% isoflurane(Ⅴ).The animals were anesthetized with intraperitoneal pentobarbital 40 mg?kg~(-1).The hearts were immediately excised and placed in cold K-H solution.The aorta was canuulated and heart retrogradely perfused with K-H solution aerated with 95% O_2 and 5% CO_2 at 37℃ and 10 kPa in a langendorff apparatus.Left ventricular end-diastollc pressure(LVEDP)and left ventrieular systolic pressure(LVSP)were measured from a fluid-filled latex balloon in the left ventricle.The isolated hearts were made globally ischemic for 30 min followed by 60 rain reperfusion in group Ⅱ-Ⅴ.In the 3 isoflurane preconditioning groups the hearts were perfused with K-H solution saturated with 0.5% or 1.0% or 2.0% isoflurane for 15 rain followed by 15 rain washout before ischemia.The cardiac function variables including LVEDP,LVSP dp/dt_(min),dp/dt_(max) and HR were measured after epuilibrium(baseline values),immediately before ischemia,at the end of 30 min ischemia and 60 min reperfusion.The infarct size and cytochrome C level in cytoplasm and mitochondria of myocytes were measured.Results I/R significantly increased LVEDP and decreased LVSP,dp/dt_(min),dp/dt_(max) as compare with control group.Sevoflurane preconditioning significantly attenuated the depression of cardiac function caused by I/R.Only LVEDP was significantly higher during reperfusion period in the 3 sevoflurane preconditioning group than in the control group but there was no significant difference in LVSP,dp/ dt_(min),dp/dt_(max) between control group and the 3 preconditioning groups.The infarct size was significantly smaller in the 3 preconditioning groups than in I/R group.Cytochrome C level was significantly increased in cytoplasm but decreased in mitochondria in I/R group as compared with control group.Sevoflurane preconditioning significantly ameliorated the release of cytochrome C from mitochondria to cytoplasm in the 3 sevoflurane preconditioning group.Conclusion Isoflurane preconditioning can protect the heart against I/R injury by attenuation of the release of cytochrone C from mitochondria to cytoplasm.