小檗碱对THP-1巨噬细胞源性泡沫细胞胆固醇流出的影响

目的 观察不同浓度小檗碱对THP-1巨噬细胞源性泡沫细胞胆固醇流出的影响,并探讨其可能的作用机制。方法 佛波酯（PMA）刺激THP-1细胞分化为巨噬细胞后经乙酰化低密度脂蛋白(AcLDL)诱导泡沫化,建立THP-1巨噬细胞源性泡沫细胞模型。根据小檗碱干预与否及干预浓度将泡沫细胞分为空白对照组和小檗碱（5~20 μmol/L）干预组。细胞经分组处理24 h后,流式细胞仪检测AcLDL聚集量;酶法检测胆固醇和乙酰甘油含量;闪烁计数法检测各组胆固醇流出率,同时分析过氧化物酶体增殖物激活受体γ（PPARγ）抑制剂GW9662预处理对胆固醇流出率的影响（以吡格列酮作为阳性对照）;RT-PCR检测肝X受体α(LXRα)和三磷酸腺苷结合盒转运体A1(ABCA1) mRNA表达。结果 与空白对照组比较,不同浓度小檗碱干预组泡沫细胞内AcLDL聚集量及胆固醇和乙酰甘油含量均显著减少（P＜0.01）,而胆固醇流出率上升（P＜0.01）,并呈现一定的量效关系。GW9662预处理后,不同浓度小檗碱干预组泡沫细胞的胆固醇流出率与对照组比较,差异均无统计学意义（P>0.05）。不同浓度小檗碱干预组泡沫细胞LXRα和ABCA1 mRNA表达均高于空白对照组。结论 小檗碱可增加THP-1巨噬细胞源性泡沫细胞胆固醇的流出量,其作用机制可能与激活PPARγ途径,增加LXRα和ABCA1 mRNA表达有关。     

褪黑素通过PPARγ-LXRα调控 THP-1巨噬细胞ABCA1表达

目的 研究褪黑素对巨噬细胞ABCA1受体表达的影响及作用机制。方法 使用THP-1细胞体外培养PMA诱导分化为巨噬细胞,给予不同浓度的褪黑素(0、50nmol/L、100nmol/L、500nmol/L和1μmol/L)干预细胞,RT-PCR和Western blot法分别检测巨噬细胞ABCA1和LXRα的mRNA和蛋白相对表达量的变化。分别应用LXRα抑制剂GSK2033、PPARγ抑制剂GW9662、非选择性MT1/MT2受体拮抗剂Luzindole、选择性MT2受体拮抗剂K185进一步验证褪黑素调控ABCA1表达的作用机制。结果 褪黑素显著上调THP-1来源巨噬细胞ABCA1和LXRα的mRNA和蛋白相对表达量,且呈剂量依赖性(P<0.05)。GW9662均可显著抑制褪黑素上调的ABCA1和LXRα蛋白表达水平(P=0.000),GSK2033可显著抑制褪黑素上调的ABCA1蛋白表达(P=0.000),但对LXRα蛋白表达水平无影响。而Luzindole和K185均未明显抑制褪黑素上调的ABCA1蛋白表达。结论 褪黑素显著上调巨噬细胞ABCA1的表达,PPARγ-LXRα通路参与了褪黑素对ABCA1表达的调节作用。     

吡格列酮对THP-1源性巨噬细胞胆固醇流出调节蛋白ABCA1,ABCG1及SR-BⅠ表达的影响

目的:探讨吡格列酮对THP-1源性巨噬细胞ATP结合盒转运蛋白A1 (ABCA1)、ATP结合盒转运蛋白G1(ABCG1)及B族Ⅰ型清道夫受体(SR-B Ⅰ)表达的影响及可能作用机制.方法:Real time PCR及Westernblot检测不同浓度吡格列酮(1,10,20,30 μmol/L)作用24 h,THP-1源性巨噬细胞ABCA1,ABCG1及SR-BⅠmRNA和蛋白的表达.使用过氧化物酶体增殖物激活型受体γ(PPARγ)特异性拮抗剂GW9662(2 μmol/L)阻断吡格列酮(10 μmol/L),24 h后检测THP-1源性巨噬细胞ABCA1,ABCG1及SR-B Ⅰ mRNA和蛋白的表达.结果:吡格列酮可浓度依赖性增高24 h THP-1源性巨噬细胞ABCA1,ABCG1及SR-BⅠ mRNA和蛋白的表达,GW9662可显著抑制吡格列酮诱导的THP-1源性巨噬细胞ABCA1,ABCG1及SR-BⅠ mRNA和蛋白表达的增高(P＜0.05).结论:吡格列酮激动PPARγ上调THP-1源性巨噬细胞胆固醇流出调节蛋白ABCA1,ABCG1及SR-BⅠ基因的表达.     

Regulation of inflammatory response in monocytes by PPARγ agonist

目的 探讨过氧化物酶体增殖物激活受体γ( PPARγ)激动剂对单核细胞炎症反应的影响.方法 体外培养THP-1人单核细胞,将细胞分为对照组、实验组、吡格列酮组和GW9662组.对照组仅加入细胞培养液;实验组用胰腺炎相关性腹水(PAAF)刺激细胞;吡格列酮组在PAAF作用前加入终浓度为20μmol/L的PPARγ激动剂吡格列酮进行干预;GW9662组:在吡格列酮干预前30 min,先行加入PPARγ拮抗剂GW9662,最后加入PAAF.分组处理6h后收集各组细胞,采用Real-Time PCR技术检测促炎症细胞因子肿瘤坏死因子α(TNF-α)和白介素6(IL-6)mRNA的表达;分别采用RT-PCR和Western blotting法检测PPARγ mRNA和蛋白的表达.结果 PAAF刺激后,与对照组比较,实验组、吡格列酮组和GW9662组细胞TNF-α和IL-6 mRNA的相对表达量明显升高,PPARγ mRNA的相对表达量明显降低,差异均有统计学意义(P＜0.05);与实验组比较,吡格列酮组TNF-α和IL-6 mRNA的相对表达量明显降低,PPARγ mRNA的相对表达量明显升高,差异均有统计学意义(P＜0.05);与吡格列酮组比较,GW9662组TNF-α和IL-6 mRNA的相对表达量均明显升高,PPARγ mRNA的相对表达量明显降低,差异也有统计学意义(P＜0.05).Western blotting检测结果显示:各组PPARγ的蛋白表达与mRNA表达的变化趋势相似,但吡格列酮组与对照组、GW9662组与吡格列酮组比较,差异均无统计学意义(P＞0.05).结论 PPARγ激动剂可通过上调PPARγ表达,减少PPAF刺激所致的促炎症细胞因子的产生,从而抑制单核细胞的炎症反应.     

PPARγ激动剂对单核细胞炎症反应的调控作用

目的探讨过氧化物酶体增殖物激活受体γ(PPARγ)激动剂对单核细胞炎症反应的影响。方法体外培养THP-1人单核细胞,将细胞分为对照组、实验组、吡格列酮组和GW9662组。对照组仅加入细胞培养液;实验组用胰腺炎相关性腹水(PAAF)刺激细胞;吡格列酮组在PAAF作用前加入终浓度为20μmol/L的PPARγ激动剂吡格列酮进行干预;GW9662组:在吡格列酮干预前30 min,先行加入PPARγ拮抗剂GW9662,最后加入PAAF。分组处理6 h后收集各组细胞,采用Real-Time PCR技术检测促炎症细胞因子肿瘤坏死因子α(TNF-α)和白介素6(IL-6)mRNA的表达;分别采用RT-PCR和Western blotting法检测PPARγmRNA和蛋白的表达。结果 PAAF刺激后,与对照组比较,实验组、吡格列酮组和GW9662组细胞TNF-α和IL-6 mRNA的相对表达量明显升高,PPARγmRNA的相对表达量明显降低,差异均有统计学意义(P0.05);与实验组比较,吡格列酮组TNF-α和IL-6 mRNA的相对表达量明显降低,PPARγmRNA的相对表达量明显升高,差异均有统计学意义(P0.05);与吡格列酮组比较,GW9662组TNF-α和IL-6 mRNA的相对表达量均明显升高,PPARγmRNA的相对表达量明显降低,差异也有统计学意义(P0.05)。Western blotting检测结果显示:各组PPARγ的蛋白表达与mRNA表达的变化趋势相似,但吡格列酮组与对照组、GW9662组与吡格列酮组比较,差异均无统计学意义(P0.05)。结论 PPARγ激动剂可通过上调PPARγ表达,减少PPAF刺激所致的促炎症细胞因子的产生,从而抑制单核细胞的炎症反应。     

脂联素对平滑肌源性泡沫细胞三磷酸腺苷结合盒转运体A1表达的影响

目的研究脂联素在体外干预泡沫细胞形成的内在机制及其对胆固醇外转运蛋白三磷酸腺苷结合盒转运体A1表达的影响。方法以平滑肌源性泡沫细胞为研究对象,采用不同浓度的脂联素体外干预平滑肌源性泡沫细胞,油红O染色观察细胞内脂滴的形成,Real-time PCR检测ABCA1转录水平的表达,Western-blot检测泡沫细胞膜蛋白LXRα、ABCA1、ABCG1和核蛋白PPARγ的磷酸化水平。结果 10μg/L脂联素能通过激活PPARγ/LXRα/ABCA1,促进胆固醇外转运蛋白ABCA1表达。结论脂联素可以激活PPARγ信号通路,促进外转运蛋白ABCA1表达,抑制平滑肌源性泡沫细胞的形成。     

桑抹茶对THP-1源性泡沫细胞脂质沉积作用的实验研究

目的探讨桑抹茶对THP-1泡沫细胞脂质沉积的改善作用。方法实验分为THP-1源性巨噬细胞对照组、THP-1泡沫细胞模型对照组、药物干预组(THP-1泡沫细胞+不同浓度桑抹茶提取物)及培养基空白对照组;采用佛波酯+Ox-LDL诱导建立THP-1泡沫细胞模型,并予不同浓度(10、20、40、80、160、320、640μg/mL)桑抹茶提取物干预,MTT法检测细胞活力;油红O染色观察细胞内脂质沉积;荧光显微镜观察THP-1巨噬细胞内吞Dil-Ox-LDL情况;Western blot检测各组细胞介导细胞内胆固醇流出蛋白ABCA1及PPARγ蛋白表达水平。结果桑抹茶提取物呈浓度依赖性抑制THP-1源性泡沫细胞对Ox-LDL的摄取,减少THP-1泡沫细胞内脂质沉积;与对照组比较,模型组泡沫细胞内脂质水平显著升高[(0.974±0.033)比(0.794±0.030),P0.01],ABCA1蛋白表达量增加[(0.702±0.005)比(0.575±0.014),P0.01],PPARγ表达量降低[(0.099±0.004)比(0.875±0.019),P0.01]。与模型组比较,93、186、372μg/mL桑抹茶提取物能显著降低泡沫细胞内脂质水平[(0.876±0.032)、(0.835±0.019)、(0.815±0.028)比(0.974±0.033),P0.01],增加PPARγ蛋白表达量[(0.270±0.006)、(0.328±0.014)、(0.706±0.020)比(0.099±0.004),P0.01],上调ABCA1蛋白表达量[(0.823±0.041)、(1.411±0.048)比(0.702±0.005),P0.01]。结论桑抹茶可能通过减少Ox-LDL摄取,促进细胞内胆固醇流出,减少细胞内脂质沉积。     

氧化芍药苷对泡沫细胞胆固醇流出的影响

分析氧化芍药苷对THP-1 巨噬细胞源性泡沫细胞胆固醇流出的影响。方法THP-1 细胞加入160 nmol/L 佛波酯（PMA）24 h，再加入50 μg/ml 乙酰化低密度脂蛋白（ac-LDL）建立泡沫细胞模型，CCK-8 法测定氧化芍药苷的细胞毒性，同位素标记法测定各处理组泡沫细胞胆固醇流出率。共聚焦显微镜观察泡沫细胞内脂肪分化相关蛋白（ADFP）表达变化，Western blot分析泡沫细胞中ADFP、三磷酸腺苷结合盒转运体A1（ABCA1）和过氧化物酶体增殖物激活受体α（PPARα）蛋白表达变化。结果成功建立THP-1 巨噬细胞源性泡沫细胞模型。氧化芍药苷毒性分析表明，质量浓度在200.0μg/ml 以下无细胞毒性。150.0μg/ml 与200.0μg/ml氧化芍药苷处理泡沫细胞，胆固醇流出率分别为（10.317±0.508）%与（11.265±0.713）%，与apoA-1组相比，差异有统计学意义（p <0.05），均高于apoA-1 组。氧化芍药苷处理的泡沫细胞内ADFP 荧光明显减弱。Western blot分析泡沫细胞ADFP 表达量降低28%。氧化芍药苷处理组PPARα 表达量增加51%，ABCA1 表达量增加45%。结论实验表明氧化芍药苷可通过上调PPARα增加ABCA1 表达，进而促进泡沫细胞胆固醇流出。     

脂多糖对小鼠巨噬细胞ABCA1表达和胆固醇流出的影响

目的观察脂多糖（LPS）对小鼠腹腔巨噬细胞ATP结合盒转运体A1（ABCA1）表达和胆固醇流出的影响,探讨肝X受体α（LXRα）信号途径的作用。方法60只C57小鼠分成两组,分别注射LPS（1.0 mg/kg）和等量生理盐水。12 h后取腹腔巨噬细胞,实时荧光定量PCR和Western-blot印迹法检测ABCA1和LXRα mRNA和蛋白质表达水平,液体闪烁计数器检测细胞内胆固醇流出。 LXRα激动剂22（R）-羟基胆固醇（22（R）-Hch）预处理小鼠,观察对LPS调节ABCA1表达和胆固醇流出的影响。结果 LPS抑制LXRα和ABCA1的mRNA和蛋白质表达;22（R）-Hch预处理抑制LPS对ABCA1表达和胆固醇流出的下调作用。结论通过抑制TLR4/LXRα信号途径的激活,可以减轻LPS诱导的急性时相反应中的炎症程度,上调ABCA1的表达,促进细胞内胆固醇的流出。     

过氧化物酶体增殖体活化受体γ、肝脏X受体α、视黄酸X受体α活化对THP-1单核巨噬细胞ABC1mRNA表达的影响及意义

目的探讨核受体过氧化物酶体增殖剂活化受体γ(peroxisome proliferator-activated receptorγ,PPARγ)、肝脏X受体α(Liver X receptor alpha,LXRα)、视黄酸X受体α(Retinoid X receptor alpha,RXRα)活化对THP-1单核巨噬细胞ATP结合盒转运蛋白(ATP binding cassette transporter 1,ABC1)mRNA表达的影响及意义。方法体外培养的THP-1单核细胞,经佛波酯(PMA)诱导成为巨噬细胞后,与核受体PPARγ、LXRα、RXRα的相应配体曲格列酮(TROG)、22(R)羟化胆固醇(22(R)-HC)、9-顺式视黄酸(9-CRA)作用24小时,提取细胞总RNA,用一步法逆转录聚合酶链反应检测核受体PPARγ、LXRα、RXRα活化剂对ABC1mRNA表达的影响。结果与对照组相比较,曲格列酮组、22(R)-HC组、9-CRA组、TROG+22(R)-HC组、TROG+组、22(R)-HC+9-CRA组ABC1 mRNA的表达均有不同程度增加,联合作用组(即核受体PPARγ、LXRα、RXRα两两活化)ABC1 mRNA增加更为明显。结论通过调控PPARγ、LXRα和RXRα等核受体的活化程度可达到调控ABC1表达的目的,这可能将为动脉粥样硬化的防治提供新的前景和机会。     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

Dr.Zhang Ren''''s Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

Luo Lingjie''''s Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.