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Endothelin-1 induces intracellular [Ca^(2+)] increase via Ca^(2+) influx through the L-type Ca^(2+) channel, Ca^(2+)-induced Ca^(2+) release and a pathway involving ET_A receptors, PKC, PKA and AT1 receptors in cardiomyocytes

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Science China(Life sciences) 2009年第4期52卷 360-370页

作者： ZENG QingHua1, LI XingTing1, ZHONG GuoGan2, ZHANG WenJie2 & SUN ChengWen3 1 Laboratory of Molecular & Cellular Physiology, school of Life sciences, Northeast Normal University, Changchun 130024, China 2 Department of Physiology, school of basic medical sciences, Jilin University, Changchun 130021, China 3 Department of Pharmaceutical sciences, North Dakota State University, Fargo, NorthDakota, USA Laboratory of Molecular & Cellular Physiology School of Life Sciences Northeast Normal University Changchun 130024 China Department of Physiology School of Basic Medical Sciences Jilin University Changchun 130021 China Department of Pharmaceutical sciences North Dakota State University Fargo NorthDakota USA

Using fura-2-acetoxymethyl ester (AM) fluorescence imaging and patch clamp techniques, we found that endothelin-1 (ET-1) significantly elevated the intracellular calcium level ([Ca2+]i) in a dose-dependent manner and activated the L-type Ca2+ channel in cardiomyocytes isolated from rats. The effect of ET-1 on [Ca2+]i elevation was abolished in the presence of the ETA receptor blocker BQ123, but was not affected by the ETB receptor blocker BQ788. ET-1-induced an increase in [Ca2+]i, which was inhibited 46.7% by pretreatment with a high concentration of ryanodine (10 μmol/L), a blocker of the ryanodine receptor. The ET-1-induced [Ca2+]i increase was also inhibited by the inhibitors of protein kinase A (PKA), protein kinase C (PKC) and angiotensin type 1 receptor (AT1 receptor). We found that ET-1 induced an enhancement of the amplitude of the whole cell L-type Ca2+ channel current and an increase of open-state probability (NPo) of an L-type single Ca2+ channel. BQ123 completely blocked the ET-1-induced increase in calcium channel open-state probability. In this study we demonstrated that ET-1 regulates calcium overload through a series of mechanisms that include L-type Ca2+ channel activation and Ca2+-induced Ca2+ release (CICR). ETA receptors, PKC, PKA and AT1 receptors may also contribute to this pathway.

关键词： endothelin-1(ET-1) cardiomyocytes intracellular calcium concentration ([Ca2+]i) L-type Ca2+ channel current (ICaL) Ca2+-induced Ca2+ release (CICR) ETA receptors PKC PKA AT1 receptors

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Cloning of Chicken Anemia Virus vp3 Gene and Apoptosis Inductive Effect of vp3 Gene In Vitro

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Journal of Huazhong University of Science and Technology(medical sciences) 2003年第4期23卷 329-331,334页

作者：孙军王宇哲宗义强屈伸 Department of Biochemistry and Molecular Biology

Using PCR technique, the vp3 gene of chicken anemia virus (CAV) was cloned into the eukaryotic expression vector pcDNA3 to construct a recombinant pcDNA vp3. Restriction enzyme digestion and sequencing analysis revealed that CAV vp3 gene was correctly inserted into the blank vector pcDNA3. After LipofectAMINE TM mediated transfection in vitro with pcDNA vp3 and pcDNA3 respectively, the total mRNA was extracted from liver carcinoma cell lines HepG2 and diploid cell line L 02, and RT PCR was performed afterward. The results of RT PCR suggested that vp3 gene was expressed in these two cell lines. At the same time, using in situ apoptotic detection assay, TUNEL kits, the apoptotic cells were found in pcDNA vp3 transfected HepG2, but not in mock transfected cell lines. VP3 could induce cell death by apoptosis in cancer cell lines, but not in diploid cell lines. All the results indicated that CAV vp3 gene, a potential therapeutic agents, has the potential of being used for cancer treatment.

关键词： chicken anemia virus gene cloning apoptosis vp3 gene

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The Effect of Neferine on Foam Cell Formation by Anti-low Density Liporotein Oxidation

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Journal of Huazhong University of Science and Technology(medical sciences) 1998年第3期18卷 134-136页

作者：冯友梅吴捷莉从容王淳本宗义强冯宗忱 DepartmentofBiochemistry SchoolofBasicMedicalSciencesTongjiMedicalUniversityWuhan.

Oxidatively modified low density lipoprtein (LDL) plays an important role in atheroslerosis (AS) development. To investigate the role of neferine (Nef) in anti-LDL oxidation and foam cell formation, the lipoprotein was derived and subjected to three different treatments: N-LDL (normal LDL), Cu(2+) +LDL and Cu(2+)+Nef+LDL. The LDLs were put at 25℃ for 24 h and the thiobarbituric acid reactive substance (TBARS) values were determined. They were 0. 57 ±0. 02, 6.01±0. 22 and 2. 26±0. 13 nmol/mg protein, respectively. The difference was very significant (P<0.01) for each two groups by t test. Mouse peritoneal macrophage (MΦ) were exposed to 50 μg protein/ml of Cu(2+) + LDL and Cu(2+)+Nef+LDL at 37℃ for 60 h. The tryglyceride (TG) and total cholesterol (TC) content in Mad were assayed. The results showed that Cu(2+) + LDL was more efficient than Cu(2+)+Nef+LDL in stimulating lipid accumulation in MΦ(P <0. 001). The study demonstrated that Nef could inhibit Cu(2+)-mediated LDL oxidation and thereby inhibiting macrophage-derived foam cell formation.

关键词： neferine low-density lipoprotein foam cell

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aFGF对肺腺癌AGZY-83A细胞株TPK、PKC活性及Ca2+浓度的影响

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中国医科大学学报 1998年第6期 15-18页

作者： Sun Liguang, Xing Wei, Liu Suyuan (Department of Biochemistry, school of basic medical sciences, China medical University, Shenyang, 110001)

目的：观察ａＦＧＦ与细胞膜上特异受体结合后引起的细胞内信号转导途径，探讨ａＦＧＦ导致细胞增殖的机理。方法：以不同浓度的ａＦＧＦ处理ＡＧＺＹ－８３Ａ细胞，利用［γ－３２Ｐ］ＡＴＰ掺入外源性底物的方法测定受体的酪氨酸蛋白... 详细信息

目的：观察ａＦＧＦ与细胞膜上特异受体结合后引起的细胞内信号转导途径，探讨ａＦＧＦ导致细胞增殖的机理。方法：以不同浓度的ａＦＧＦ处理ＡＧＺＹ－８３Ａ细胞，利用［γ－３２Ｐ］ＡＴＰ掺入外源性底物的方法测定受体的酪氨酸蛋白激酶活性（ＴＰＫ）及蛋白激酶Ｃ（ＰＫＣ）活性；用Ｆｕｒａ－２／ＡＭ为荧光指示剂测定［Ｃａ２＋］ｉ。结果：随着ａＦＧＦ浓度增加，ＴＰＫ及ＰＫＣ活性随之升高。当ａＦＧＦ浓度为１．１２μｇ／ｍｌ时ａＦＧＦ处理组的ＴＰＫ是对照组的４倍；膜ＰＫＣ活性也是对照组的４倍，胞浆ＰＫＣ活性是对照组的１．７５倍。［Ｃａ２＋］ｉ是对照组的３倍。结论：该细胞株中ａＦＧＦ受体具有ＴＰＫ活性。ＴＰＫ激活后进一步促进蛋白质和酶磷酸化，而使ＰＫＣ活性及［Ｃａ２＋］ｉ升高，即ＰＫＣ和Ｃａ２＋是ＴＰＫ的下游信号分子，进一步促进ｃ－ｆｏｓ、ｊｕｎ基因表达增加，导致细胞增殖

关键词：酪氨酸蛋白激酶蛋白激酶C 胞浆Ca2+浓度细胞增殖

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Differential Expression of NDAP Elevates Living Threshold via ER Stress

Differential Expression of NDAP Elevates Living Threshold vi...

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药物相关蛋白质组学专题研讨会

作者： Yu chang hai school of basic medical sciences, Peking University, Beijing 100083 China

Switching on and off key molecules during life or death decisions, plays fundamental roles in brain morphogenesis. NDAP ( neuronal development - associated protein) was previously identified by our laboratory and shown to be differentially expressed during development and regulated by neurotrophins in culture. In this study, we found that an increase in NDAP was correlated with apoptosis during development. We demonstrated that NDAP - EGFP was an ER resident protein. Overexpression of NDAP - EG-

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Effects of acetazolamide and anordiol on osmotic water permeability in AQP1-cRNA injected Xenopus oocyte

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Acta Pharmacologica Sinica 2004年第1期 92-99页

作者： Bing MA, Yang XIANG, Sheng-mei MU, Tao LI, He-ming YU, Xue-jun LIDepartment of Pharmacology, school of basic medical sciences, Peking University, Beijing 100083 National Research Institute for Family Planning, Beijing 100081, China

AIM: To study the effects of acetazolamide and anordiol on osmotic water permeability in aquaporin 1 (AQP1)-cRNA injected Xenopus oocyte and their mechanisms. METHODS: AQP1 gene constructed in pBluescript was transcripted into cRNA in vitro and then the cRNA was injected in Xenopus oocytes. The effects of acetazolamide and anordiol on the water transport function of AQP1 were observed by assaying the osmotic swelling of oocytes. In addition, their effects on protein expression of AQP1 were quantitatively investigated by Western blotting method. RESULTS: After incubation for 15 min or 72 h, acetazolamide, a carbonic anhydrase inhibitor, equally reduced the water permeability of AQP1-cRNA injected oocyte in a dose-dependent manner. After incubation for 72 h, anordiol, an antiestrogen with partial estrogenic activity, reduced the osmotic water permeability dose dependently as well; however, no discernable action was observed after incubation with anordiol for 15 min. The Western blotting

关键词： aquaporins acetazolamide anordiol permeability Xenopus

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The Pharmacological Study of Lingzhi and the Research of the Therapeutical Principle of Fuzheng Guben in Traditional Chinese Medicine

The Pharmacological Study of Lingzhi and the Research of the...

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2002国际灵芝专题研讨会

作者： Zheng Cong Department of Pharmacology, school of basic medical sciences , Peking University, Beijing, China

This paper reviewed the historical course of the research work of Lingzhi in author's laboratory. They proposed a working hypothesis to guide their pharmacological studies of Chinese tonic drugs and the Therapeutical Principle of Fuzheng Guben in TCM and introduced their successful experiences. On this basis, the author suggested some strategic considerations which may be useful in the research of Chinese traditional drugs.

关键词： Lingzhi, Ganoderma lucidum , traditional Chinese medicine (TCM), the Therapeutical Principle of Fuzheng Guben in TCM, tonic drugs, working hypothesis

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Effects of U-50 488H a k-agonist on action potentials of isolated ventricular papillary muscle of guinea pigs

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Acta Pharmacologica Sinica 1994年第1期15卷 37-39页

作者： SHENG Jian-Zhong, DONG Hui, WONG Tak-Ming(Department of Physiology, Faculty oj Medicine, The University oj Hong Kong, Hong Kong) Recipient of a fellowship from Sun Yat Sen Foundation, now in Department of Physiology, Zhejiang Meidical University, Hangzhou 310006, China.Department of Biophysics, school of basic medical sciences, Shanghai medical University, Shanghai 200032, China. Department of Physiology Faculty of Medicine University of Hong Kong Hong Kong Hong Kong

Trans (+-)-3,4-dichloro-N-methyl-N-[ 2-( 1-pyrrolidinyl ) cyclohexyl ]-benzeneacetamide-methanesulfonate-hydrate (U-50 488H), a specific K-agonist. at 1 - 10 μmol·L-1 caused concentration-dependent reductions in the action potential duration at 5.0% and 90% of repolarization (APD50 and APD90) without modifying the resting potential (RP), the action potential amplitude (APA) and the maximal upstroke velocity (Vmax). The effects were attenuated by (- )- (lR,5R.9R)-5,9-diethyl-2-(3-furylmethyI)-2 L - hydroxy- 6 , 7 - benzomorphan (Mr 2266 BS, 1 μmol·L-1), a specific K-antagonist which itself had no effect on the action potentials of the ventricular papillary muscle of guinea pigs, indicating that U-50 -188H at 1-10 μmol·L-1 acts via specific cardiac K-receptors. At 100 μmol·L-1, U-50 488H not only shortened APD50 and APD90 but also reduced RP, APA, and Vmax which were not attenuated by Mr 2266 BS (1 μmol·L-1) suggesting that the effects of U-50 488H at 100 μmol · L-1 were probably nonspecific.

关键词： U-50 488H Mr 2266 BS endorphin receptors action potentials papillary muscles guinea pigs

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Effects and possible mechanisms of habenular nucleus in the pathogenesis of obstructive sleep apnea

Effects and possible mechanisms of habenular nucleus in the ...

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2006年中国睡眠研究会第四届学术年会

作者： Hua Zhao (Department of Physiology,school of basic medical sciences, Ji Lin University, Changchun 130021)

The morphological and physiological studies have indicated an important role for serotonin in the central mechanism of OSAS occurring. The reduction of 5-HT in the nuclei of raphe suppressed the activities of hypoglossal motor neurons during sleep ,then pharynx dilator muscle relaxed and resulted in OSAS at last. Habenular nucleus(Hb) located in thalamencephalon projected directly to the nuclei of raphe and influenced its functions. The expeirment aim is to investigate the regulating effect of Hb on the nuclei of raphe in OSAS and find the possiblly referred neurotransmitters or mediator. The results are stated as

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Pathogenicity of GB virus C on virus hepatitis and hemodialysis patients

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World Journal of Gastroenterology 2003年第8期9卷 1739-1742页

作者： Wan-Fu Zhu Li-Min Yin Peng Li Jian Huang Hui Zhuang, Department of Microbiology, school of basic medical sciences, Peking University, Beijing 100083, China Wan-Fu Zhu Li-Min Yin Peng Li Jian Huang Hui Zhuang Department of Microbiology School of Basic Medical Sciences Peking University Beijing 100083 China

AIM: To determine the pathogenicity of GB virus C (GBV-C)on liver and the effects of its co-infection on the clinical features and prognosis of patients with hepatitis B and ***: Cross-sectional study was carried out in 413patients with acute, chronic hepatitis B or liver cirrhosis,and in 67 hemodialysis patients. A 20-month prospective cohort study was carried out in 95 hepatitis B and 80 hepatitis C patients. A reverse transcriptase nested polymerase chain reaction (RT-nPCR) of the 5′-noncoding region was used to detect circulating GBV-C RNA. Liver function was determined by an automated analyzer for all ***: The prevalence of GBV-C in the high-risk populations with the virus transmitted via blood was high,ranging from 16.2 to 28.8%. Co-infection with GBV-C in hepatitis B patients did not affect the clinical features of the disease or liver function. The dialysis patients infected with GBV-C alone did not develop functional changes to the *** cohort study showed that GBV-C co-infection did not affect the clinical features, prognosis or negative serum conversion rate of chronic hepatitis B and ***: The results suggest that GBV-C has no marked pathogenicity on liver, so it may not be a hepatitis virus.

关键词：病毒性肝炎血液透析 GB病毒病原学

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