大鼠肾俞定位之我见

肾俞主治病症多,临床应用广泛,因此对于肾俞穴的实验研究层出不穷。大鼠作为实验常用动物,其腧穴的准确定位对于针灸实验研究具有重要的意义。本研究在阐明大鼠脊椎解剖特点的基础上,指明大鼠肾俞的定位方法,希望对针灸实验研究能起到一定的指导作用。

电针、埋线对小鼠吗啡戒断和耐受调整作用的对比研究

目的通过对吗啡戒断、耐受小鼠进行电针和穴位埋线治疗,观察小鼠海马和脊髓中N-甲基-D-天冬氨酸(N-methyl-D-aspartate,NMDA)受体和胆囊收缩素(cholecystokinin,CCK)的表达水平,探讨电针和埋线对吗啡戒断后戒断、耐受小鼠调整作用的差异。方法将56只雄性C57BL/6J小鼠随机分为戒断空白组、戒断模型组、戒断埋线组、戒断电针组和耐受空白组、耐受模型组、耐受埋线组、耐受电针组,每组7只。吗啡戒断模型根据"7 d递增成瘾法"皮下注射盐酸吗啡注射液,戒断空白组在相同时间点注射等量生理盐水,戒断电针组小鼠每次注射盐酸吗啡注射液15 min后,使用韩式穴位神经刺激仪(HANS-200)电针刺激双侧肾俞穴;戒断埋线组于注射盐酸吗啡注射液15 min后,将0.5 cm铬肠线埋入双侧肾俞穴内。第7天早上10点,腹腔注射盐酸纳洛酮注射液催瘾(4 mg/kg),并观察小鼠戒断反应。根据阿片依赖戒断症状柳田知司测评量表评分,并以酶联免疫吸附剂测定法(Elisa)检测海马和脊髓NMDA受体和CCK含量。按10 mg/kg皮下注射吗啡建立吗啡耐受模型。耐受空白组在相同的时间注射10 m L/kg的生理盐水,耐受埋线组于建立模型第1天在肾俞穴埋线治疗,耐受电针组在建立模型第1天起电针肾俞穴。经7 d治疗后,以Elisa方法检测海马和脊髓的NMDA受体和CCK含量。结果戒断模型组海马NR2B和CCK表达与戒断空白组比较,差异均具有统计学意义(P<0.05)。戒断电针组海马NR2B表达与戒断模型组比较,差异有统计学意义(P<0.05)。戒断埋线组和戒断电针组海马CCK表达与戒断模型组比较,差异均具有统计学意义(P<0.05)。戒断模型组小鼠脊髓NR2A、NR2B、CCK表达与戒断空白组比较,差异均具有统计学意义(P<0.05)。戒断电针组脊髓NR2A、NR2B表达与戒断模型组比较,差异均具有统计学意义(P<0.05)。耐受模型组海马NR1、NR2B、CCK表达与耐受空白组比较,差异均具有统计学意义(P<0.05)。耐受埋线组海马CCK表达与耐受模型组比较,差异具有统计学意义(P<0.05)。耐受电针组海马NR1表达与耐受模型组和耐受埋线组比较,差异均具有统计学意义(P<0.05)。耐受埋线组和耐受电针组脊髓CCK表达与耐受模型组比较,差异均具有统计学意义(P<0.05)。结论肾俞穴穴位埋线及电针治疗对减轻吗啡耐受及戒断均有治疗效果,且电针治疗优于穴位埋线。

艾灸“肾俞”早期干预去卵巢合D-半乳糖注射AD样大鼠效应中的特异性

Objective:To observe the effect of the early intervention with moxibustion at "Shènshū(肾俞BL23)" on learning and memory of Alzheimer’s disease(AD) rats modeled by ovariectomy and D-Galactose(D-Gal)injection and the effect on Ca2+/CaMKII/CREB signaling pathway so as to provide the evidence for the acupoint selection in the early intervention with acupuncture and moxibustion for AD.Methods:Sixty female SD rats at the age of 3 months were selected and randomly divided into a normal group,a sham-operation group,a model group,a "BL23" group,a "Wèishū"(胃俞BL21) group,and a non-acupoint group,10 rats in each group.Except in the normal group and the sham-operation group,bilateral ovaries were removed in the other 4 groups.3 days after the operation,D-Gal was injected intraperitoneally at a dose of 150 mg/kg/d for 90 days to establish AD rat model.In the sham-operation group,only the peripheral fat of ovarian was removed and 0.9% normal saline was injected intraperitoneally of the same dose for 90 days.In the "BL23" group,the "BL21" group and the non-acupoints group,since the 2 nd day after ovariectomy,moxibustion was applied at 9:00 a.m.every day in the rats.The rats were fixed on a fixed table,the mild moxibustion with self-prepared moxa stick was applied to bilateral "BL23""BL21" and the non-acupoint of each rat,the ignited end of moxa stick was 2 cm to 3 cm far from skin surface.The infrared thermometer was adopted to measure the temperature of the epidermis of each acupoint from time to time so as to maintain the local temperature to be(41 ± 0.5)℃.Mild moxibustion lasted for 10 min each time,consecutively for 5 days a week and at an interval of 2 days.The total consecutive intervention duration was 12 weeks.In the normal group and the shamoperation group,no moxibustion intervention was performed,but the rats were restricted in a same way during the experiment.At the end of modeling and intervention,water maze test was adopted to detect the learning and memory abilities of rats in each group.The western blotting was used to detect the expressions of p-CREB,p-CaMKII,CaMKII and CREB in hippocampus of rats in each group.Results:(1)Navigation test:since Day 72,compared with the sham-operation group,the escape latencies in all of the consecutive 5 days were longer in the model group(all P <0.05).Since Day 73,compared with the normal group,the escape latency in all of the consecutive 4 days was shorter in the "BL23"group(all P <0.05).(2) Times across platform:compared with the normal group,the number of times consecutively crossing the platform was lower in the model group(P <0.05).After moxibustion,compared with the model group,the number of times across platform was higher in the "BL23"group,indicating a statistical significance(P <0.05).(3)Moving track of water maze spatial probe:the "tendency"strategy was displayed in the normal group and the sham operation group.The "marginal" strategy,which is commonly seen in the AD rats,was displayed in the model group.The "random" strategy was displayed in the "BL23" group.The "marginal" strategy was presented in the "BL21" group and the non-acupoint group.(4) Expressions of hippocampal P-CAMKII,CAMKII,CREB and P-CREB in the rats:compared with the normal group,the phosphorylation degrees of CAMKII and CREB were significantly reduced(both P<0.05) in the model group.In 12 weeks of moxibustion,compared with the model group,the levels of P-CAMKII and P-CREB were increased in the "BL23" group and the "BL21" group(all P <0.05) and the levels in the "BL23" group were higher than that of the "BL21" group and the non-acupoint group(all P<0.05).Conclusion:Moxibustion at "BL23" achieves a better effect in improving the learning and memory of AD rats modeled by ovariectomy and D-Gal injection as compared with moxibustion at "BL21" and the non-acupoint.The effect mechanism is potentially related to the effective activation of Ca2+/CaMKII/CREB signaling pathway.

Difference in the anti-fatigue effect of moxibustion at different acupoints in one-off exhausted rats

Objective:To explore the difference in the anti-fatigue effect of moxibustion at different acupoints through observing the impacts of moxibustion at"Shènshù(肾俞BL23)","Zúsānlǐ(足三里ST36)","Guānyuán(关元CV4)"and"Shénquè(神阙CV8)"on energy metabolic indexes in the one-off exhausted rats.Methods:A total of 56 SPF male rats were randomized into 7 groups,named a blank group,a model group,a non-acupoint group,a"BL23"group,a"ST36"group,a"CV4"group and a"CV8"group,8 rats in each group.The one-off exhaustion model was established by weight-bearing swimming experiment.Right after the exhaustive swimming of rats in each intervention group,the mild moxibustion was applied at the corresponding acupoints for 15 min immediately.4 h after the exhaustive swimming,the blood was collected from the femoral artery and the quadriceps femoris of the left hindlimb was stripped.The levels of blood lactate(BLD),lactate dehydrogenase(LDH),creatine kinase(CK)and blood urea nitrogen(BUN),the expressions of adenosine triphosphatease(ATP),fructose-6-phosphofructokinase(PFK)and malomdialdehyde(MDA)in the quadriceps femoris and the activity of superoxide dismutase(SOD)and catalase(CAT)were detected.Results:Compared with the model group and the non-acupoint group,the levels of serum BLD,LDH,CK and BUN were all lower those in the"BL23"group,the"ST36"group,the"CV4"group and the"CV8"group,indicating a statistical significance(all P<0.05).Compared with the model group and the nonacupoint group,PFK was higher in the"BL23"group,indicating a statistical significance(both P<0.05).Compared with the model group and the non-acupoint group,PFK was higher in the"ST36"group and the"CV4"group,indicating a statistical significance(all P<0.05).Conclusion:It was found in the research that moxibustion at"BL23""ST36""CV4"and"CV8"regulated the energy metabolism in the one-off exhausted rats so as to bring the anti-fatigue effect into play and the effect was different among the acupoints.Moxibustion at"CV8"may work on the prevention of the protein decomposition in the skeletal muscle and moxibustion at"BL23"may be against the oxidation in the skeletal muscle.