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Isoform-selective modulators for metabotropic glutamate receptors and protein kinase c:Synthesis and biological evaluation

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chinese Journal of chemistry 1998年第4期16卷 292-302页

作者：马大为 Shanghai Institute of Organic chemistry Chinese Academy of SciencesShangha 200032China

The synthetic studies for some known modulators of metabotropic glutamate receptors (mGluRs) such as (S)-αM4cPG, (1S,3R)-AcPD, L-ccG-I are described. Based on the structure of αM4cPG several new conformationally constrained analogues are design ed and synthesized. Among them APIcA is a selective antagonist for group II mGluRs. Also, a new benzolactam-V8 analogue is found to have better isoform-selectivity for protein kine c family. Three different protocols for synthesizing benzolactam-VS analogues are developed to meet the requirement for delivering more analogues to test.

关键词： Asymmetric synthesis metabotropic glutamate receptor protein kinase c

Ginsenosides stimulated the proliferation of mouse spermatogonia involving activation of protein kinase c

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2009年第2期10卷 87-92页

作者： Da-lei ZHANG Kai-ming WANG cai-qiao ZHANG Key Laboratory of Animal Epidemic Etiology and Immunological Prevention of the Ministry of Agriculture College of Animal Sciences Zhejiang University Hangzhou 310029 China college of Medicine Nanchang University Nanchang 330006 China

The effect of ginsenosides on proliferation of type A spermatogonia was investigated in 7-day-old mice. Spermatogonia were characterized by c-kit expression and cell proliferation was assessed by immunocytochemical demonstration of proliferating cell nuclear antigen (PcNA). After 72-h culture, Sertoli cells formed a confluent monolayer to which numerous spermatogonial colonies attached. Spermatogonia were positive for c-kit staining and showed high proliferating activity by PcNA expression. Ginsenosides (1.0～10 μg/ml) significantly stimulated proliferation of spermatogonia. Activation of protein kinase c (PKc) elicited proliferation of spermatogonia at 10-8 to 10-7 mol/L and the PKc inhibitor H7 inhibited this effect. Likewise, ginsenosides-stimulated spermatogonial proliferation was suppressed by combined treatment of H7. These results indicate that the proliferating effect of ginsenosides on mouse type A spermatogonia might be mediated by a mechanism involving the PKc signal transduction pathway.

关键词： Ginsenosides Spermatogonia protein kinase c Mouse

calycosin improves cognitive function in a transgenic mouse model of Alzheimer's disease by activating the protein kinase c pathway

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Neural Regeneration Research 2017年第11期12卷 1870-1876页

作者： Lei Song Xiaoping Li Xiao-xue Bai Jian Gao chun-yan Wang Department of Respiratory Medicine First Hospital of Jilin UniversityChangchunJilin ProvinceChina Department of Pediatrics First Hospital of Jilin UniversityChangchunJilin ProvinceChina cadre's Ward First Hospital of Jilin UniversityChangchunJilin provinceChina Department of Neurosurgery China-Japan Union Hospital of Jilin UniversityChangchunJilin ProvinceChina

The major pathological changes in Alzheimer＇s disease are beta amyloid deposits and cognitive impairment. calycosin is a typical phy- toestrogen derived from radix astragali that binds to estrogen receptors to produce estrogen-like effects. Radix astragali calycosin has been shown to relieve cognitive impairment induced by diabetes mellitus, suggesting calycosin may improve the cognitive function of Alzhei- mer＇s disease patients. The protein kinase c pathway is upstream of the mitogen-activated protein kinase pathway and exerts a neuropro- tective effect by regulating Alzheimer＇s disease-related beta amyloid degradation. We hypothesized that calycosin improves the cognitive function of a transgenic mouse model of Alzheimer＇s disease by activating the protein kinase c pathway. Various doses of calycosin （10, 20 and 40 mg/kg） were intraperitoneally injected into APP/PS1 transgenic mice that model Alzheimer＇s disease. calycosin diminished hippocampal beta amyloid, Tau protein, interleukin-lbeta, tumor necrosis factor-alpha, acetylcholinesterase and malondialdehyde levels in a dose-dependent manner, and increased acetylcholine and glutathione activities. The administration of a protein kinase c inhibitor, cal- phostin c, abolished the neuroprotective effects of calycosin including improving cognitive ability, and anti-oxidative and anti-inflammato- ry effects. Our data demonstrated that calycosin mitigated oxidative stress and inflammatory responses in the hippocampus of Alzheimer＇s disease model mice by activating the protein kinase c pathway, and thereby improving cognitive function.

关键词： nerve regeneration neurodegeneration Alzheimer's disease calycosin hippocampus oxidative stress inflammation mice protein kinase c calphostin c glutathione malondialdehyde neural regeneration

Fungus induces the release of IL- 8 in human corneal epithelial cells, via Dectin-1-mediated protein kinase c pathways

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International Journal of Ophthalmology(English edition) 2015年第3期8卷 441-447页

作者： Xu-Dong Peng Gui-Qiu Zhao Jing Lin Nan Jiang Qiang Xu cheng-cheng Zhu Jian-Qiu Qu Lin cong Hui Li Department of Ophthalmology the Affiliated Hospital of Qingdao University

AIM: To identify whether Aspergillus fumigatus(***) hyphae antigens induced the release of interleukin-8(IL-8) in anti-fungal innate immunity of cultured human corneal epithelial cells(HcEcs) and determine the involvement of intracellular signalling pathways. METHODS: HcEcs were treated with A. fumigatus hyphae antigens with different concentrations and *** cytoplasmic calcium of HcEcs were assessed by fluorescence imaging. Western blot was used to detect the expression of ca2 +-dependent protein kinase c(PKc). The IL-8 levels were determined by specific human IL-8 enzyme-linked immunosorbent assay(ELISA) and reverse transcriptase polymerase chain reaction(RT-PcR). Using a series of pharmacological inhibitors, we examined the upstream signalling pathway responsible for IL-8 expression in response to *** hyphae antigens. RESULTS: cells exposed to A. fumigatus hyphae antigens showed higher level of IL-8 m RNA expression and protein production. We demonstrated here that stimulation of HcEcs with A. fumigatus hyphae triggers an intracellular ca2 +flux and results in the activation of ca2 +-dependent PKc(α, βⅠ and βⅡ) which can be attenuated by pre-treatment of cells with laminarin,suggesting that Dectin-1 signals pathway induced cytoplasmic calcium and influence the activation of PKc in HcEcs. Inhibitors of ca2 +-dependent PKc(Ro-31-8220 and Go6976) significantly abolished hyphae-induced expression of ***: Our findings suggest that A. fumigatushyphae-induced IL-8 expression was regulated by the activation of Dectin-1-mediated ca2 +-dependent PKc in HcEcs.

关键词： Dectin-1 ca 2+ protein kinase c interleukin-8 corneal epithelium Aspergillus fumigatus

Effects of Mitochondrial ATP-sensitive K^+ channel on protein kinase c Pathway and Airway Smooth Muscle cell Proliferation in Asthma

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2012年第4期32卷 480-484页

作者：万璇赵建平谢俊刚 Department of Pulmonary and critical care Medicine Tongji HospitalTongji Medical CollegeHuazhong University of Science and Technology

The effects of ATP-sensitive mitochondrial K + channel(mitoK ATP) on mitochondrial membrane potential(Δψm),cell proliferation and protein kinase c alpha(PKcα) expression in airway smooth muscle cells(ASMcs) were ***-six Sprague-Dawley(SD) rats were immunized with saline(controls) or ovalbumin(OVA) with alum(asthma models).ASMcs were cultured from the lung of control and asthma *** were treated with diazoxide(the potent activator of mitoK ATP) or 5-hydroxydencanote(5-HD,the inhibitor of mitoK ATP).Rhodamine-123(R-123) was used to detect Δψ*** expression of PKcα protein was examined by using Western blotting,while PKcα mRNA expression was detected by using real-time *** proliferation of ASMcs was measured by MTT assay and cell cycle *** diazoxide-treated normal ASMcs,the R-123 fluorescence intensity,protein and mRNA levels of PKcα,MTT A values and percentage of cells in S phase were markedly increased as compared with untreated *** ratio of G 0 /G 1 cells was decreased(Pcs from normal ***,there were no significant differences between the ASMcs from healthy rats treated with 5-HD and the normal control *** untreated and diazoxide-treated ASMcs of asthmatic rats,the R-123 fluorescence intensity,protein and mRNA levels of PKcα,MTT A values and the percentage of cells in S phase were increased in comparison to the normal control ***,in comparison to ASMcs from asthmatic rats,these values were considerably increased in asthmatic group treated with diazoxide(Pcreased as compared with asthma control group(Pcs of asthma,the signal transduction pathway of PKcα may be involved in cell proliferation,which is induced by the opening of mitoK ATP and the depolarization of Δψm.

关键词： asthma airway smooth muscle cells ATP-sensitive K + channel protein kinase c

An Experimental Study on the Regulation of Expression of Th_2 cytokines from T Lymphocytes by protein kinase c in Asthma

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2001年第4期21卷 292-296页

作者：熊维宁徐永健张珍祥王孝养莫碧文傅娟 Department of Respiratory Medicine Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030

To explore the regulatory role of protein kinase c (PKc) in the expression of Th 2 cytokines, interleukin 4 (IL 4) and interleukin 5 (IL 5) by T lymphocytes in asthma. T lymphocytes were isolated and purified from blood and bronchial alveolus lavage fluid (BALF) of each guinea pig of normal control group and asthmatic group and from peripheral blood of the asthmatic patients and normal controls, and were stimulated with PKc accelerant phorbol 12 myristate 13 acetate (PMA) and inhibitor Ro31 8220. The expression of IL 4 and IL 5 mRNA and protein was detected by using in situ hybridization staining and ELISA respectively. The expression of IL 4 and IL 5 mRNA and protein of asthmatic T lymphocytes stimulated with PMA was significantly higher than that of asthmatic T lymphocytes stimulated without PMA respectively ( P cytes stimulated with PMA respectively ( P protein of asthmatic T lymphocytes stimulated with PMA and Ro31 8220 was significantly lower than that of asthmatic T lymphocytes stimulated only with PMA respectively ( P concluded that PKc might participate in regulating the expression of IL 4 and IL 5 in asthmatic T lymphocytes, and the activation of PKc in T lymphocytes might play an important role in the pathogenesis of asthma.

关键词： protein kinase c bronchial asthma interleukin 4 interleukin 5

Regulations of Thyroid Hormone on cardiac protein kinase c Signal Pathway in vitro

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Wuhan University Journal of Natural Sciences 2004年第4期9卷 522-526页

作者： WANGBao-hua OUYANGJing-ping LIUYong-min ZHENGHan-qiao LIYin-ping SchoolofMedicine WuhanUniversityWuhan430071HubeiChina SchoolofMedicine WuhanUniversityWuhan430071HubeiChina

The experiments were conducted to assess the influences of thyroid hormone on cardiac protein kinase c(PKc) signal pathway with cultured cardiac myocytes and fibroblasts as the models. cells were pretreated with 1% newborn calf serum (NcS) or angiotensin II (Ang II), and then following by a triiodothyronine (T3) treatment. The PKc activity, PKcα and PKcε expressions were analyzed and compared. In 1% NcS pretreatment, T3 could inhibit PKc activity and PKcε expression in cardiac myocytes. The AngII pretreatment led to an increase of PKc activity and PKcε expression in cardiac myocytes, and an increase of PKc activity in cardiac fibroblasts. Following by T3 treatment, the increased PKc activity and PKcε expression in cardiac myocytes were markedly decreased. In conclusion, whether in 1% NcS or in Ang II pretreatment, T3 could inhibit PKc activity and PKcε expression in cardiac myocytes. Key words thyroid hormone - cardiac myocytes - cardiac fibroblasts - protein kinase c cLc number Q 572 Foundation item: Supported by the Natural Science Foundation of Hubei Province (98091)Biography: WANG Bao-hua (1974-), female, Ph. D, research direction: cardiovascular pathophysiology.

关键词： thyroid hormone cardiac myocytes cardiac fibroblasts protein kinase c

The changes of protein kinase c Activity in Peripheral Blood Lymphocytes in the Patients with Obstructive Jaundice and the Implication

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2001年第2期21卷 119-121页

作者：王剑明邹倩邹声泉 Department of Surgery Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030

The roles of protein kinase c signal pathway in the pathogenesis of obstructive jaundice were studied. PKc from cytosolic and membrane fractions of peripheral blood lymphocytes in 51 patients with obstructive jaundice and 16 cases of normal controls was isolated and purified. The activities of PKc were determined by radioactive isotope γ 32 P ATP catalyzing assay. The results showed that the total PKc activities in PBL in the patients with obstructive jaundice were significantly increased as compared with those in the normal controls . Moreover, the membrane PKc activities and their percentages of the total PKc activities were higher in obstructive jaundice group than in those in the normal controls . The total PKc activities in PBL in the patients with obstructive jaundice were significantly positively correlated with the levels of soluble IL 2 receptor and the degree of jaundice in serum. It was concluded that the activities of PKc signal pathway was related with the degree of T BIL. PKc signal pathway might took part in the activation of T lymphocytes in the patients with obstructive jaundice and play an important role in the immune regulation and the assessment of pathosis in the patients with obstructive jaundice.

关键词： obstructive jaundice lymphocytes protein kinase c

protein kinase c regulates neurite outgrowth in spinal cord neurons

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Neuroscience Bulletin 2010年第2期26卷 117-125页

作者：杨萍李振强宋林殷玉芹第三军医大学解剖学教研室重庆400038 第三军医大学神经生物学教研室重庆400038 哈佛医学院儿童医院神经外科神经科学实验室波士顿02115

Objective The functional roles of protein kinase c （PKc） in the neurite outgrowth and nerve regeneration remain controversial. The present study was aimed to investigate the role of PKc in neurite outgrowth, by studying their regulatory effects on neurite elongation in spinal cord neurons in vitro. Methods The anterior-horn neurons of spinal cord from embryonic day 14 （E14） Sprague-Dawley （SD） rats were dissociated, purified and cultured in the serum-containing medium. The ratio of membrane-PKc （mPKc） activity to cytoplasm-PKc （cPKc） activity （m/c-PKc） was studied at different time points during culture. Results Between 3-11 d of culture, the change of m/c-PKc activity ratio and PKc-βⅡ expression in the neurite were both significantly correlated with neurite outgrowth （r=0.95, P 〈 0.01; r=0.73, P 〈 0.01, respectively）. Moreover, PMA, an activator of PKc, induced a dramatic elevation in the m/c-PKc activity ratio, accompanied with the increase in neurite length （r=-0.99, P 〈 0.01）. In contrast, GF 109203X, an inhibitor of PKc, significantly inhibited neurite elongation, which could not be reversed by PMA. conclusion PKc activity may be important in regulating neurite outgrowth in spinal cord neurons, and βⅡ isoform of PKc probably plays a major role in this process.

关键词： protein kinase c spinal cord neurons neurite outgrowth in vitro