黄芪总苷对脾虚大鼠胃粘膜TFF1和EGF表达的影响

目的:探讨黄芪总苷对冷水-束缚应激状态下脾虚大鼠胃粘膜TFF1、EGF mRNA和蛋白表达的影响。方法:大鼠70只分为7组,即正常对照组、脾虚组、模型(脾虚应激)组、黄芪总苷(高剂量200mg/kg、中剂量100mg/kg、低剂量50mg/kg)组、补中益气丸3.7g/kg组。采用大黄致脾虚法复制脾虚模型大鼠,水浸-束缚法给予脾虚大鼠应激,酶联免疫(ELISA)法检测TFF1、EGF蛋白表达变化,RT-PCR法测定TFF1、EGF的mRNA表达变化。结果:与正常组比较,脾虚组大鼠胃粘膜TFF1 mRNA和蛋白表达下降,EGF mRNA低表达,而脾虚应激组TFF1、EGF mRNA和蛋白表达增加;与脾虚应激组相比,补中益气丸组和黄芪总苷(200mg/kg)组TFF1和EGF表达均明显增加。结论:黄芪总苷对脾虚大鼠胃粘膜的保护作用机制可能与影响TFF1和EGF有关。     

Effect of moxibustion on CRF and CRFR1 expressions in hypothalamus of TNBS-induced experimental colitis rats

Objective

To observe the effect of moxibustion on the protein and mRNA expressions of corticotropin-releasing factor (CRF) and corticotropin-releasing factor receptor 1 (CRFR1) in hypothalamus of trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis rats, and to explore the central mechanisms of moxibustion in improving visceral pain and the pain-related emotions in experimental colitis rats.

Methods

Thirty-six Sprague-Dawley (SD) rats were randomly divided into a normal group (NG), a model group (MG), a herb-partitioned moxibustion group (HPMG) and a sham herb-partitioned moxibustion group (SHPMG). Except the NG, rats in the remaining three groups all received TNBS enema to establish experimental colitis models. The HPMG received herb-partitioned moxibustion (HPM) at bilateral Tianshu (ST 25) and Qihai (CV 6) for intervention; for the SHPMG, the herbal cakes and moxa cones were only placed on the acupoints but not ignited; rats in the MG and NG were only fixed in the same way as those in the HPMG but did not receive any treatment. At the end of the intervention, the abdominal withdrawal reflex (AWR) score, the open field test (OFT) score and the elevated plus maze (EPM) score were observed to measure the changes in visceral pain and pain-related emotions of the rats. The enzyme-linked immunosorbent assay (ELISA) was used to examine the expressions of CRF and CRFR1 proteins in hypothalamus; the fluorescence-based quantitative polymerase chain reaction (PCR) was used to detect the expressions of CRF and CRFR1 mRNAs in hypothalamus.

Results

Compared with the NG, the AWR score increased significantly and the OFT and EPM scores dropped significantly in the MG (all P<0.05), and the expressions of hypothalamic CRF and CRFR1 proteins and mRNAs increased significantly (all P<0.01). Compared with the MG and SHPMG, the AWR score dropped significantly and the OFT and EPM scores increased significantly in the HPMG (all P<0.01), and the expressions of hypothalamic CRF and CRFR1 proteins and mRNAs decreased significantly (all P<0.05). There were no significant differences between the MG and the SHPMG (all P>0.05).

Conclusion

HPM can down-regulate the abnormally increased expressions of CRF and CRFR1 proteins and mRNAs in hypothalamus of the TNBS-induced experimental colitis rats, which is plausibly one of its action mechanisms in mitigating visceral pain and the pain-related emotions in the experimental colitis rats.

     

Effects of different doses of ginger-partitioned moxibustion on trefoil factor 1, mucin 5AC and epidermal growth factor receptor in rats with spleen deficiency syndrome

Objective

To observe the effects of different doses of ginger-partitioned moxibustion on serum trefoil factor 1 (TFF1) and mucin 5AC (MUC5AC) levels, as well as the expression of epidermal growth factor receptor (EGFR) in gastric mucosa of rats with spleen deficiency syndrome, therefore, to explore the possible mechanism and the dose-effect characteristics of ginger-partitioned moxibustion in spleen deficiency syndrome.

Methods

Seventy-five SPF grade Sprague-Dawley (SD) rats were randomly divided into a blank control group (group A), a model group (group B), a 3 moxa-cone ginger-partitioned moxibustion group (group C1), a 6 moxa-cone ginger-partitioned moxibustion group (group C2) and a 9 moxa-cone ginger-partitioned moxibustion group (group C3) using random number table method, 15 rats in each group. Except group A, rats in the other groups received intragastric administration of 4 °C 200% concentrated Da Huang (Radix et Rhizoma Rhei) to prepare spleen deficiency syndrome model. After successful modeling, rats in group B received no treatment; rats in group C1, C2 and C3 were treated with 3, 6 and 9 moxa-cone ginger-partitioned moxibustion at Zusanli (ST 36) and Zhongwan (CV 12) respectively for 8 continuous days. The general symptom score of rats was observed. The serum levels of TFF1 and MUC5AC were detected by enzyme-linked immunosorbent assay (ELISA). The expression of EGFR protein in gastric mucosa was detected by immunohistochemistry.

Results

After the treatment, compared with group A, the spleen deficiency symptom score was increased in group B, the levels of serum TFF1 and MUC5AC, the EGFR protein expression in gastric tissues of group C1, C2 and C3 were significantly increased (all P<0.01); compared with group B, the spleen deficiency scores were decreased in group C1, C2 and C3, and the serum levels of TFF1 and MUC5AC, as well as EGFR protein expression in gastric tissues were increased (all P<0.01). Compared with group C1, the spleen deficiency scores were decreased in group C2 and C3, the serum levels of TFF1 and MUC5AC, and the expression of EGFR protein in gastric tissues were increased (all P<0.01), however, there was no significant difference between group C2 and C3 (all P>0.05). The mechanism may be related to the increase of serum TFF1 and MUC5AC levels and activation of EGFR protein.

Conclusion

Ginger-partitioned moxibustion can improve the symptoms, as well as promote the proliferation and repair of gastric mucosa in rats with spleen deficiency. The therapeutic efficacy of 6 or 9 moxa-cone ginger-partitioned moxibustion is better than that of 3 moxa-cone ginger-partitioned moxibustion, while the efficacies are equivalent between 6 and 9 moxa-cone ginger-partitioned moxibustion groups.

     

Thirty-eight cases of uterine bleeding of spleen deficiency treated with wheat-grain size cone moxibustion and herbal plaster on the umbilicus

Objective:To introduce an effective method in treatment of uterine bleeding of spleen deficiency.Methods:A total of 38 patients were treated with wheat-grain size cone moxibustion and herbal plaster on the umbilicus.The wheat-grain size cone moxibustion was used at Guānyuán(关元 CV 4)and Yǐn bái(隐白 SP 1),20 moxa cones at each acupoint were required in each treatment.The treatment was given once every two days and three treatments made 1 course.Additionally,the herbal plaster was dressed on the umbilicus.The dressing changed once every day.After 2 courses of treatment,the therapeutic effect was observed.Results:Regarding the effects of uterine bleeding,of 38 cases,12 cases were cured,accounting for 31.58%,17 cases effective remarkably,accounting for 44.74%,4 cases effective,accounting for 10.53% and 5 cases failed,accounting for 13.16%.The total effective rate was 86.84%.The symptoms of spleen deficiency were relieved in 38 patients,such as sallow complexion,lassitude,short breathing,dislike of speaking,no appetite and cold limbs.Conclusion:The combined treatment with wheat-grain size cone moxibustion and herbal plaster on the umbilicus achieves significant effects on uterine bleeding of spleen deficiency.     

艾灸对溃疡性结肠炎大鼠结肠KGF-1、KGF-2和IL-6蛋白表达的影响

目的:观察艾灸对溃疡性结肠炎大鼠结肠角质细胞生长因子-1 (Keratinocyte Growth Factor-1,KGF-1)、角质细胞生长因子-2(Keratinocyte Growth Factor-2,KGF-2) KGF-2和白介素-6(Interleukin-6,IL-6)蛋白表达的影响,进一步探讨艾灸治疗溃疡性结肠炎的作用机制.方法:将SD大鼠随机分为正常组、模型组、隔药灸组和柳氮磺胺吡啶(Sulfasalazine,SASP)组.隔药灸组采用隔药灸天枢和气海穴治疗;SASP组采用柳氮磺胺吡啶灌胃干预.干预结束后,应用HE染色光镜下观察各组大鼠结肠病理学变化,采用免疫组织化学法检测大鼠结肠KGF-1、KGF-2和IL-6蛋白表达变化.结果:与模型组比较,隔药灸组和SASP组大鼠结肠病理学均有一定的改善;与正常组比较,模型组大鼠结肠KGF-1、KGF-2和IL-6蛋白表达均显著增加(P＜0.05);经隔药灸和SASP干预后,大鼠结肠KGF-1、KGF-2和IL-6蛋白表达均显著降低(P＜0.05).结论:隔药灸和SASP均能下调溃疡性结肠炎大鼠结肠KGF-1、KGF-2和IL-6的蛋白表达,该作用可能是隔药灸和SASP治疗溃疡性结肠炎的重要机制之一.     

艾灸对卵巢储备功能减退大鼠Nrf2/HO-1信号通路的影响

目的:观察艾灸对卵巢储备功能减退(DOR)大鼠核因子E2相关因子2(Nrf2)/血红素加氧酶1(HO-1)信号通路的影响,探讨艾灸对卵巢储备功能的保护效应机制。方法:将40只SD大鼠随机分为空白组、模型组、艾灸组、激素组,每组10只。模型组、艾灸组、激素组均采用雷公藤多苷片混悬液灌胃制备DOR大鼠模型,空白组灌胃等体积的0.9%氯化钠溶液,每日1次,连续灌胃14 d。激素组在造模即日起采用激素序贯疗法连续干预14 d;艾灸组在造模即日起每日给予双侧"肾俞"或"关元""中脘"艾灸治疗,两组穴隔日交替,每次10 min,连续14 d。每日采用阴道脱落细胞涂片观察大鼠的动情周期,统计各组大鼠动情周期紊乱率。干预结束后,采用HE染色观察各组大鼠卵巢组织学形态;ELISA法检测各组大鼠血清卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇(E2)、抗苗勒管激素(AMH)和超氧化物歧化酶(SOD)、丙二醛(MDA)含量;免疫组化法检测各组大鼠卵巢组织Nrf2和HO-1蛋白水平;实时PCR(TaqMan探针法)检测各组大鼠卵巢组织Nrf2和HO-1 mRNA的表达。结果:与空白组比较,模型组大鼠动情周期紊乱率升高(P<0.01);与模型组比较,艾灸组和激素组大鼠动情周期紊乱率降低(P<0.01)。与空白组比较,模型组大鼠血清FSH、LH、MDA含量增多(P<0.01),血清E2、AMH、SOD含量减少(P<0.01);与模型组比较,艾灸组和激素组大鼠血清FSH、LH、MDA含量减少(P<0.01,P<0.05),血清E2、AMH、SOD含量增多(P<0.01)。与空白组比较,模型组大鼠卵巢组织Nrf2和HO-1蛋白及mRNA表达减少(P<0.01);与模型组比较,艾灸组和激素组大鼠卵巢组织Nrf2和HO-1蛋白及mRNA表达增多(P<0.01)。结论:艾灸可以降低DOR大鼠的动情周期紊乱率,改善血清性激素水平和抗氧化应激能力,其机制可能与调控Nrf2/HO-1信号通路有关。     

肝郁脾虚证胃溃疡大鼠胃窦组织差异蛋白质表达及柴黄胃溃宁的干预研究

目的:探讨肝郁脾虚证胃溃疡大鼠胃窦组织总蛋白质的差异表达及柴黄胃溃宁的可能疗效机制。方法:以多因素复合模拟中医病因结合乙酸法建立肝郁脾虚证胃溃疡大鼠模型,采用双向凝胶电泳技术分析模型大鼠与正常大鼠胃窦组织总蛋白质的表达差异;同时,采用笔者临床经验方柴黄胃溃宁对模型大鼠进行复健。结果:模型组大鼠的血清胃泌素、血清淀粉酶活性、血浆去甲肾上腺素和血浆5-羟色胺等指标出现紊乱,与正常组比较,差异均有统计学意义(P<0.05,P<0.01);与正常组匹配后,模型组5个蛋白质点表达下调,2个蛋白质点表达上调;经柴黄胃溃宁治疗后,大鼠上述血清、血浆指标复健;模型组和中药组匹配后,模型组5个蛋白点表达低于中药组,1个点表达高于中药组。结论:与正常组的蛋白质可能是肝郁脾虚证胃溃疡的相关蛋白;柴黄胃溃宁对肝郁脾虚证胃溃疡的疗效可靠,其作用机制可能与调控上述目的蛋白质有关。     

毫火针配合隔物艾灸治疗脾胃虚寒型功能性消化不良的临床疗效

[目的] 探讨毫火针配合隔物艾灸治疗脾胃虚寒型功能性消化不良的临床疗效。[方法] 选择2022年9月—2023年9月珠海市中西医结合医院收治的90例脾胃虚寒型功能性消化不良患者,随机分为两组。对照组（45例）采用毫火针治疗,观察组（45例）采用毫火针配合隔物艾灸治疗。比较症状评分,血清胃肠激素、炎症因子,生活质量,疗效和安全性。[结果] 两组治疗后症状评分较治疗前降低（P＜0.01）;肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-6、IL-1β水平较治疗前降低（P＜0.01）,血清促胃液素（GAS）、胃动素（MTL）较治疗前增高（P＜0.01）;健康查简表（SF-36）评分均较治疗前增高（P＜0.01）。观察组治疗后血清TNF-ɑ、IL-6、IL-1β水平低于对照组（P＜0.01）,GAS、MTL高于对照组（P＜0.01）;SF-36评分高于对照组（P＜0.01）。观察组治疗总有效率高于对照组（77.78% vs. 93.33%,P＜0.05）。[结论] 毫火针配合隔物艾灸治疗更有助于改善胃肠动力,抑制炎症反应,提高脾胃虚寒型功能性消化不良的临床疗效和生活质量。     

电针配合隔药灸治疗女性脾肾阳虚型单纯性肥胖28例

目的:观察电针配合隔药灸治疗脾肾阳虚型女性单纯性肥胖的临床疗效.方法:将58例患者随机分为针灸组(28例)和电针组(30例).电针组单纯采用电针治疗,穴取中脘、下脘、气海、关元、天枢、大横、减肥点、阿是穴等,其中天枢配大横、减肥点配下脘和气海行电针治疗,留针40 min.针灸组在电针基础上,在脾俞和肾俞穴加用隔药饼灸.5次为一疗程,3个疗程后评定疗效.结果:针灸组总有效率为92.9％( 26/28)优于电针组的70.0％(21/30,P＜0.05).结论:电针配合隔药灸治疗脾肾阳虚型女性单纯性肥胖能提高疗效,优于单纯电针治疗.     

黄芪总苷对脾虚大鼠胃黏膜保护机制探讨

目的:探讨黄芪总苷对冷水-束缚应激状态下脾虚大鼠胃黏膜H+,K+-ATP酶活性及mRNA表达变化、胃蛋白酶活性及髓过氧化物MPO活性的影响。方法:SD大鼠70只分为7组,即正常对照组,脾虚组,模型(脾虚应激)组,黄芪总苷(200mg/kg、100mg/kg、50mg/kg)组,补中益气丸组。采用经典大黄致脾虚法复制脾虚模型大鼠,水浸-束缚法给予脾虚大鼠应激,观察胃黏膜损伤指数,无机磷法检测胃黏膜H+,K+-ATP酶、胃蛋白酶及MPO活性的变化,RT-PCR法测定H+,K+-ATP酶mRNA表达变化。结果:正常对照及脾虚组大鼠胃黏膜未见溃疡损伤,经冷水-束缚应激后的模型组、补中益气丸组、黄芪总苷(高、中、低)组大鼠胃黏膜可见点状或条索状溃疡损伤,且与模型组相比黄芪总苷中剂量及高剂量组、补中益气丸组明显降低(P<0.01)。与正常组比较,模型组大鼠胃黏膜H+,K+-ATP酶活性、胃蛋白酶及髓过氧化物MPO活性值,H+,K+-ATP酶mRNA表达明显增高(P<0.05),而脾虚组降低(P<0.05)。与模型组相比,补中益气丸组、黄芪总苷(中、高)剂量组H+,K+-ATP酶活性、胃蛋白酶及髓过氧化物MPO活性值,H+,K+-ATP酶mRNA表达均有所下降(P<0.05)。结论:黄芪总苷对脾虚大鼠胃黏膜的保护作用机制之一与影响胃蛋白酶、H+,K+-ATP酶活性及其mRNA表达有关。     

香砂六君子汤对脾气虚证浅表性胃炎大鼠线粒体能量代谢影响研究

目的:探讨香砂六君子汤对脾气虚证浅表性胃炎模型大鼠体内的琥珀酸脱氢酶活性与细胞色素氧化酶C含量的作用。方法:采用劳倦过度与饥饱失常加强酸刺激等复合因素塑造脾气虚证模型,在此基础上运用强酸刺激加感染幽门螺旋杆菌法复制大鼠脾虚浅表性胃炎模型,观察香砂六君子汤对脾气虚证浅表性胃炎大鼠胃窦组织琥珀酸脱氢酶活性和细胞色素氧化酶C含量的影响。结果:模型组大鼠胃窦细胞琥珀酸脱氢酶活性显著低于正常组和治疗组,差异具有统计学意义(P<0.05),治疗组琥珀酸脱氢酶活性显著高于模型组,差异具有统计学意义(P<0.05),治疗组与正常组比较,差异无统计学意义(P>0.05); 正常组大鼠胃窦组织细胞色素氧化酶C含量明显高于模型组,差异具有统计学意义(P<0.05),治疗组胃窦细胞色素氧化酶C含量高于模型组,差异具有统计学意义(P<0.05),治疗组与正常组比较,差异无统计学意义(P>0.05)。结论:香砂六君子汤治疗脾气虚证浅表性胃炎的效果确切,作用机制可能与其提高胃窦线粒体琥珀酸脱氢酶活性与细胞色素氧化酶C含量相关。     

Regulating effects of herb cake-partitioned moxibustion on the expression of p53 and C-myc protein in rats with ulcerative colitis

目的：从p53基因、原癌基因蛋白质C—myc表达与溃疡性结肠炎发病关系角度,探讨隔药灸对p53、C—myc表达影响,进一步探讨隔药灸治疗UC作用机制。方法：采用免疫学方法加局部刺激制备溃疡性结肠炎大鼠模型。40只SD大鼠随机数字表法分为正常组、模型组、隔药灸组、温和灸组,每组10只。温和灸组,选取天枢、气海穴悬灸,每次每穴灸10min。隔药灸组选取天枢、气海穴隔药灸,每次每穴灸2壮。免疫组织化学法检测溃疡性结肠炎大鼠结肠组织中p53、C—myc表达。结果：模型组大鼠结肠组织中p53、C-myc的表达在阳性面积、强度、免疫组织化学（immunohistochemistry,mc）指数方面均显著上调,与正常组比较具有显著差异（P〈0．01）;隔药灸组、温和灸组治疗后结肠Np53、C—myc的表达下调,与模型组比较具有显著差异（P〈0．01）结论：p53与C．myc在UC发生、发展中具有重要作用,隔药灸对UC结肠组织p53、C-myc表达水平具有调节作用。     

痹肿消汤拆方对胶原诱导性关节炎大鼠滑膜ERK1/2、ERK5表达及其转导通路的影响

目的:检测痹肿消汤按湿、热、毒、瘀拆方对胶原诱导性关节炎大鼠滑膜细胞外信号调节激酶1/2(ERK1/2)、细胞外信号调节激酶5(ERK5)表达的影响,从湿、热、毒、瘀探讨类风湿性关节炎(RA)的作用机制。方法:采用皮下注射牛Ⅱ型胶原蛋白与完全弗氏佐剂诱导SD大鼠胶原诱导性关节炎(CIA)模型,并设立正常组。采用免疫印迹法(Western blot)同步检测正常组、模型组、痹肿消汤组、清热解毒组(拆方Ⅰ组)、祛湿组(拆方Ⅱ组)、活血组(拆方Ⅲ组)大鼠在不同时间点滑膜ERK1/2、ERK5的磷酸化水平,运用灰度扫描软件对结果进行半定量分析。结果:造模14d后,模型组滑膜ERK1/2、ERK5表达高于正常组(P0.05)。随着免疫时间延长,模型组ERK1/2、ERK5表达于21、28d呈递增趋势(P0.05),28d达到最高。4用药组治疗后,ERK1/2、ERK5的磷酸化表达水平均低于同期模型组(P0.05)。在21、28d水平,表达呈递减趋势(P0.05)。痹肿消汤组的ERK1/2、ERK5各时间点的表达水平比拆方3组更低,拆方Ⅱ组的ERK1/2表达的下降幅度比拆方I组、拆方Ⅲ组更明显(P0.05)。而拆方I组的ERK5表达的下降幅度比拆方Ⅱ组、拆方Ⅲ组更显著(P0.05)。结论:拆方各组可有效地抑制ERK1/2、ERK5转导通路的活化,达到阻抑关节滑膜增殖、骨质侵蚀的作用。但清热解毒、祛湿通络、活血祛瘀各治法对上述通路调节作用优势有不同。     

不同形式阴阳调理灸治疗脾虚湿阻型单纯性肥胖症疗效观察

目的观察不同形式阴阳调理灸治疗脾虚湿阻型单纯性肥胖症的临床疗效,对具体实施方法进行优选。方法将96例脾虚湿阻型单纯性肥胖症患者按随机数字表法分为3组。3组均予饮食运动干预及常规针刺治疗,隔药饼灸组32例(后退出1例)采用隔药饼灸治疗,温和灸组32例(后退出1例)采用温和灸治疗,温灸器灸组32例采用温灸器治疗。比较3组治疗前后中医证候评分、腰围(WC)、体质量(BW)、体质量指数(BMI)、体脂肪率(F%)的差异,并统计疗效。结果隔药饼灸组愈显率67. 7%(21/31),温和灸组愈显率38. 7%(12/31),温灸器灸组愈显率40. 6%(13/32),隔药饼灸组愈显率优于温和灸组和温灸器灸组(P <0. 05)。治疗后3组WC、BW、BMI、F%值均较本组治疗前降低(P <0. 05),且治疗后隔药饼灸组WC、BW、BMI、F%值均低于温和灸组和温灸器灸组(P <0. 05)。隔药饼灸组治疗后各项中医证候评分均较本组治疗前降低(P <0. 05),温和灸组和温灸器灸组治疗后水肿、脘腹满闷、大便稀溏评分均较本组治疗前降低(P <0. 05),且治疗后隔药...     

Effects of moxibustion pretreatment on extracellular signal-regulated kinase signaling transduction pathway in the gastric tissues of rats with gastric mucosal damage

Objective

To observe the effects of moxibustion pretreatment on the protein expressions of epidermal growth factor receptor (EGFR), phosphorylation extracellular signal-regulated kinase 1/2 (p-ERK1/2) and activated protein-1 (AP-1), the key factors of extracellular signal-regulated kinase signaling transduction pathway in gastric tissue of rats with stress-induced gastric mucosal damage, and to discuss the mechanisms of moxibustion therapy in promoting the restoration of damaged gastric mucosa.

Methods

Thirty Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group, and a moxibustion group using the random digits table, 10 in each group. Except the rats in the normal group, rats in the other two groups were used to make stress-induced gastric mucosal damage model using restraint and cold stress. Before modeling, rats in the moxibustion group were alternately treated with moxibustion at Zusanli (ST 36) and Zhongwan (CV 12), or Pishu (BL 20) and Weishu (BL 21), once a day, for a total of 8 d. Histological changes of gastric mucosa were observed under the light microscopy, the expression of gastric tissue p-ERK1/2 was detected by immunohistochemistry assay, and the protein levels of EGFR and AP-1 were measured by Western blots.

Results

Compared with rats in the normal group, gastric mucosal damage was more serious, and protein expressions of gastric tissue EGFR, p-ERK1/2 and AP-1 increased in the model group (P<0.01, P<0.05, P<0.05). Compared with rats in the model group, gastric mucosal damage was milder, and protein expressions of gastric tissue EGFR, p-ERK1/2 and AP-1 increased in the moxibustion group (all P<0.01).

Conclusion

Moxibustion at Zusanli (ST 36), Zhongwan (CV 12), Pishu (BL 20) and Weishu (BL 21) could increase EGFR, p-ERK1/2 and AP-1 expression levels in gastric tissue of stress-induced gastric mucosal damage rats, maintain the information transfer function of ERK signaling transduction pathway, and promote restoration of gastric mucosal damage.

     

黄芪皂苷干预脾虚水湿不化证大鼠的代谢组学研究

目的研究黄芪皂苷组分干预脾虚水湿不化证大鼠血浆内源性代谢物的变化,探求黄芪皂苷组分对脾虚水湿不化证的干预作用。方法采用高效液相-飞行时间质谱仪检测大鼠血浆内源性代谢物信息,利用主成分分析和偏最小二乘判别分析研究对照组、模型组以及黄芪皂苷组之间的代谢差异,确定潜在生物标志物。结果对照组、模型组以及黄芪皂苷组代谢模式明显不同,共鉴定11个生物标志物,其通路主要涉及甘油磷脂代谢、鞘脂代谢、淀粉与蔗糖代谢和花生四烯酸代谢等。结论黄芪皂苷组分可能通过干预溶血磷脂酰胆碱导致的脂肪代谢紊乱、脂肪酸进入线粒体减少导致的能量供给不足、肠道功能失调等一系列代谢紊乱对脾虚水湿不化证起到治疗作用。     

Effect of moxibustion on survival status and nutritional metabolic factor in tumor-bearing rats with gastric cancer

Objective:To observe the effect of moxibustion on the survival status and nutritional metabolic factors of tumor-bearing rats with gastric cancer and explore the inhibitory effect of moxibustion on the tumorbearing rats.Methods:A total of 40 SD rats were randomly divided into a blank group,a sham-operated group,a model group and a moxibustion group,10 rats in each group.Gastric cancer models were established in the model group and the moxibustion group.The rats of all the groups underwent the same constraining procedure for 20 min every day.Additionally,moxibustion was applied at acupoints for 20 min in the rats of the moxibustion group.There were two groups of acupoints.One group included"Zusanli(足三里ST 36)""Zhongwan(中脘CV12)"and"Guanyuan(关元CV4)".The other group included bilateral"Pishu(脾俞BL 20)"and"Weishu(胃俞BL 21)".Moxibustion was applied for 20 min every day at the acupoints of the two groups alternatively and lasted for 14 days.The survival status of rats was observed and the score of survival status and body weight of rats were recorded every day.At the end of intervention,the orbital blood was collected and the rats were sacrificed for sample collection.The viscera-free body weight was recorded.Using biochemical analyzer,the blood glucose(GLU),albumin(ALB),total protein(TP)and triglyceride(TG)in serum were detected.Results:(1)Regarding the score of the survival status,after modeling,the scores in the model group and the moxibustion group were all higher than those of the sham-operation group(both P0.05).After intervention,the score in the moxibustion group was lower than that of the model group(P0.05).(2)After modeling,the body weight in the moxibustion group and the model group was lower than those of the blank group,indicating a statistical significance(both P0.05).The body weight in the moxibustion group was higher than that of the model group after intervention,indicating the statistical significance(P0.05).(3)Regarding the viscera-free body weight,it was lower in the model group compared with the sham-operation group,indicating the statistical significance(P0.05).Compared with the model group,the viscera-free body weight was higher in the moxibustion group,indicating the statistical significance(P0.04).(4)Regarding the nutritional metabolic factors,compared with the sham-operation group,the levels of GLU and ALB in the model group were lower and TP was higher,presenting the statistical significance(all P0.05).Compared with the model group,the levels of GLU and ALB were higher in the moxibustion group and TP was lower,presenting the statistical significance(all P0.05).Conclusion:Moxibustion improves the survival status of tumor-bearing rats with gastric cancer,increases the body weight,the viscera-free body weight and the levels of GLU and ALB in serum and reduces TP of the tumor-bearing rats with gastric cancer and inhibits the growth of gastric cancer.     

补肾健脾中药复方对去势大鼠血清中Bcl-2和Bax水平的影响

目的:探讨补肾健脾中药复方对去势大鼠血清中Bcl-2和Bax水平的影响。方法:48只6月龄的雌性SD大鼠随机分为对照组、模型组、中药组(补肾健脾中药复方)、西药组(雌激素)。治疗12周后,收集左侧股骨、血清分别行骨矿含量、骨密度测量和Bcl-2、Bax水平含量的检测。结果:模型组的骨密度值和骨矿含量均低于对照组(P0.05);中药组和西药组的骨密度值高于模型组(P0.05)。与空白组相比,模型组血清中Bcl-2表达水平下降,Bax表达水平上升(P0.05)。经过药物干预后,中药组和西药组均能提高血清中Bcl-2表达,降低Bax表达(P0.05)。结论:补肾健脾中药复方对去势大鼠血清中Bcl-2和Bax水平的影响作用与雌激素一致,有可能在一定程度上抑制细胞的凋亡。     

参归仁合剂对产后抑郁大鼠行为学及雌激素介导下ERK1/2通路中MEK1的影响

目的:探讨参归仁合剂对产后抑郁模型大鼠行为学的影响,及测其海马、额叶、下丘脑神经元内胞外信号调节激酶1/2(ERK1/2)通路上丝裂原活化细胞外调节激酶1(MEK1)蛋白表达量。方法:150只雌性大鼠分为正常组,模型组,参归仁高、中、低剂量组(7,3.6,1.8 g·kg-1·d-1),阳性药组(舍曲林4.5 mg·kg-1·d-1和苯甲酸雌二醇0.01 mg·kg-1·d-1),除正常组外,其余各组ih苯甲酸雌二醇和黄体酮造成抑郁模型。观察其行为学指标变化;蛋白质免疫印迹(Western blot)法检测其额叶、海马、下丘脑中p-MEK1,MEK1的蛋白表达。结果:与正常组比较,模型组体重、食欲、活动能力、兴趣等明显降低,额叶、海马、下丘脑细胞内的p-MEK1蛋白表达显著升高(P0.01);参归仁合剂能改善抑郁模型组体重、食欲、活动能力、兴趣等行为学指标,抑郁模型组额叶、海马、下丘脑细胞内的p-MEK1,MEK1蛋白表达量明显下降,参归仁高、中、低组额叶、海马、下丘脑细胞内的p-MEK1,MEK1蛋白表达量显著增加,尤其以参归仁中、低组增加较为明显(P0.01)。结论:参归仁合剂能改善产后抑郁大鼠的行为学指标,在一定程度上调ERK1/2信号通路p-MEK1,MEK1的蛋白表达量,且中、低组效果显著,提示该药作用机制为调控ERK1/2通路并改善抑郁症状。     

基于MEK/ERK信号通路探讨阳和汤含药血清对乳腺癌4T1细胞的影响

目的 通过阳和汤含药血清干预小鼠乳腺癌4T1细胞,探索阳和汤对乳腺癌4T1细胞及其内丝裂原活化蛋白激酶（MEK）/细胞外调节蛋白激酶（ERK）信号通路的影响。方法 制备阳和汤SD大鼠含药血清。将其随机分组为空白组、阳和汤低、中、高剂量组（5.8、11.6、23.2 g·kg^-1）,空白组用生理盐水代替。各组按10%含药血清和90% RMPI 1640完全培养基均匀混合,干预细胞。采用细胞增殖与活性检测（CCK-8）法分别在24、48、72 h检测阳和汤含药血清对4T1细胞增殖的影响;采用流式细胞术检测4T1细胞的凋亡情况;采用划痕实验检测4T1细胞迁移情况;采用Transwell实验检测细胞侵袭能力;采用蛋白免疫印迹法（Western blot）检测MEK1/2、磷酸化（p）-MEK1/2、ERK1/2、p-ERK1/2、大鼠肉瘤病毒（RAS）蛋白的表达水平。结果 与空白组比较,阳和汤干预24、48、72 h,阳和汤含药血清低、中、高剂量组对4T1细胞增殖具有明显抑制作用（P<0.05,P<0.01）;阳和汤干预48 h,阳和汤含药血清低、中、高剂量组细胞凋亡率显著升高（P<0.01）;阳和汤含药血清低、中、高剂量组在24、48 h的细胞划痕愈合能力显著降低（P<0.01）;阳和汤含药血清低、中、高剂量组细胞侵袭能力显著降低（P<0.01）;阳和汤含药血清低、中、高剂量组细胞内MEK1/2、ERK1/2表达差异无统计学意义,p-MEK1/2、p-ERK1/2、RAS蛋白表达显著降低（P<0.01）。结论 阳和汤可抑制乳腺癌细胞4T1增殖、迁移、侵袭,促进其凋亡。其凋亡机制与MEK/ERK信号通路受抑制,p-MEK1/2、p-ERK1/2、RAS蛋白表达下调有关。