Parkinson＇s disease （PD） is the most common motor neurode- generative disorder affecting approximately 4 million people worldwide. Although PD presents primarily with motor dysfunction, non-motor symptoms including...Parkinson＇s disease （PD） is the most common motor neurode- generative disorder affecting approximately 4 million people worldwide. Although PD presents primarily with motor dysfunction, non-motor symptoms including cognitive decline, mood disorders, reduced olfaction and constipation are also of- ten present, with some of these non-motor symptoms even pre- senting prior to the onset of motor symptoms. It is well known that PD is largely caused by the gradual degeneration of dopa- minergic neurons within the substantia nigra pars compacta （SNc）, along with the presence of protein aggregates called Lewy bodies, which consist primarily of ct-synuclein and are found in the cytoplasm of surviving neurons. This ongoing cell loss and Lewy body pathology is not confined to the SNc, but is also seen in other brain regions implicated in PD pathogenesis such as the locus ceruleus.展开更多
OBJECTIVE To study the potential relationship between the expression of PPT-I, NK-1, NK2 and the development of breast cancer cells in bone marrow stroma and to provide evidence of potential molecular mechanisms of br...OBJECTIVE To study the potential relationship between the expression of PPT-I, NK-1, NK2 and the development of breast cancer cells in bone marrow stroma and to provide evidence of potential molecular mechanisms of breast cancer patients. of bone metastasis in early stage METHODS The cocultures of breast cancer cell line T-47D and marrow-derived mesenchymal stem cells （MSC） were established with equal numbers. T-47D cells were separated from the coculture system at 48 h and 96 h after coculture by MACS magnetic cell sorting （MicroBeads）. The expression of PPT-I, NK-1, NK-2 in T-47D was then examined before and after coculture by real-time PCR and by Western blot. Alterations in cellular ultrastructure of T-47D cells were detected before and after coculture under electron microscope. Finally, changes in cell cycle distribution were examined by flow cytometry, and growth curves from before and after coculture were drawn and analyzed. RESULTS Following coculture of T-47D and MSC, the expression of PPT-I mRNA and protein was significantly upregulated, while the expression of NK-1 and NK-2 mRNA and protein was greatly downregulated. The analysis of cell cycle distribution by flow cytometry showed that the proportion of T-47D during S phase was increased, and the duration of the G2/M phase was sharply decreased. Under electron microscope, we observed that the synthesis of hereditary material was increased, but the hepatin granules were shown prominent stacking in T-47D cells. These results suggest that although the synthesis of DNA was increased, the proliferation of cells was inhibited after coculture. The cell growth curve confirmed the findings from the observation under the electron microscope and flow cytometry. CONCLUSION Tumor cells could survive through the upregulation in expression of preprotachykinin-I gene during early bone metastasis in breast cancer. The phenomenon of growth suppression in breast cancer cells after coculture in the current study could be induced by downregulation in expression of NK-1 and NK-2.展开更多
The expression of the types of tachykinin receptors in the dorsal root ganglion (DRG) neurons by means of Xenopus oocyte expressing system was studied. Poly(A) +RNAs were extracted from cat cervical and lumbar DRG. Tw...The expression of the types of tachykinin receptors in the dorsal root ganglion (DRG) neurons by means of Xenopus oocyte expressing system was studied. Poly(A) +RNAs were extracted from cat cervical and lumbar DRG. Two days after injection of Poly(A) +RNAs, the oocytes were recorded with the two electrode voltage clamp technique. In the oocytes injected with DRG poly(A) +RNA,[Sar 9,Met(O 2) 11 ] substance P(Sar SP, 1 μmol/L), neurokinin A (NKA,1 μmol/L) or [β Ala 8] neurokinin A (4-10) (Ala NKA, 1 μmol/L) produced an inward current comprising a rapid spike and a long sustained oscillatory component for several minutes. Sar SP induced response was blocked by NK 1 antagonist L 668, 169 (1 μmol/L), but not by NK 2 antagonist L 659,877(1μmol/L). In contrast, Ala NKA and NKA responses were only blocked by L 659,877. The oocytes injected with DH Poly(A) +RNA also responded to Sar SP and NKA with similar inward currents, which were selectively blocked by L 668,169 and L 659,877, respectively. These tachykinins induced responses had a potent desensitization. The present data indicate expression of NK 1 and NK 2 receptors in DRG neurons, suggesting that there may be tachykinin autoreceptors on the nociceptive primary afferent terminals.展开更多
基金in part has been supported by the Neurosurgical Research Foundation, South Australia, Australia
文摘Parkinson＇s disease （PD） is the most common motor neurode- generative disorder affecting approximately 4 million people worldwide. Although PD presents primarily with motor dysfunction, non-motor symptoms including cognitive decline, mood disorders, reduced olfaction and constipation are also of- ten present, with some of these non-motor symptoms even pre- senting prior to the onset of motor symptoms. It is well known that PD is largely caused by the gradual degeneration of dopa- minergic neurons within the substantia nigra pars compacta （SNc）, along with the presence of protein aggregates called Lewy bodies, which consist primarily of ct-synuclein and are found in the cytoplasm of surviving neurons. This ongoing cell loss and Lewy body pathology is not confined to the SNc, but is also seen in other brain regions implicated in PD pathogenesis such as the locus ceruleus.
基金supported by a grant from the National Natural Science Foundation of China(No.30670802).
文摘OBJECTIVE To study the potential relationship between the expression of PPT-I, NK-1, NK2 and the development of breast cancer cells in bone marrow stroma and to provide evidence of potential molecular mechanisms of breast cancer patients. of bone metastasis in early stage METHODS The cocultures of breast cancer cell line T-47D and marrow-derived mesenchymal stem cells （MSC） were established with equal numbers. T-47D cells were separated from the coculture system at 48 h and 96 h after coculture by MACS magnetic cell sorting （MicroBeads）. The expression of PPT-I, NK-1, NK-2 in T-47D was then examined before and after coculture by real-time PCR and by Western blot. Alterations in cellular ultrastructure of T-47D cells were detected before and after coculture under electron microscope. Finally, changes in cell cycle distribution were examined by flow cytometry, and growth curves from before and after coculture were drawn and analyzed. RESULTS Following coculture of T-47D and MSC, the expression of PPT-I mRNA and protein was significantly upregulated, while the expression of NK-1 and NK-2 mRNA and protein was greatly downregulated. The analysis of cell cycle distribution by flow cytometry showed that the proportion of T-47D during S phase was increased, and the duration of the G2/M phase was sharply decreased. Under electron microscope, we observed that the synthesis of hereditary material was increased, but the hepatin granules were shown prominent stacking in T-47D cells. These results suggest that although the synthesis of DNA was increased, the proliferation of cells was inhibited after coculture. The cell growth curve confirmed the findings from the observation under the electron microscope and flow cytometry. CONCLUSION Tumor cells could survive through the upregulation in expression of preprotachykinin-I gene during early bone metastasis in breast cancer. The phenomenon of growth suppression in breast cancer cells after coculture in the current study could be induced by downregulation in expression of NK-1 and NK-2.
文摘The expression of the types of tachykinin receptors in the dorsal root ganglion (DRG) neurons by means of Xenopus oocyte expressing system was studied. Poly(A) +RNAs were extracted from cat cervical and lumbar DRG. Two days after injection of Poly(A) +RNAs, the oocytes were recorded with the two electrode voltage clamp technique. In the oocytes injected with DRG poly(A) +RNA,[Sar 9,Met(O 2) 11 ] substance P(Sar SP, 1 μmol/L), neurokinin A (NKA,1 μmol/L) or [β Ala 8] neurokinin A (4-10) (Ala NKA, 1 μmol/L) produced an inward current comprising a rapid spike and a long sustained oscillatory component for several minutes. Sar SP induced response was blocked by NK 1 antagonist L 668, 169 (1 μmol/L), but not by NK 2 antagonist L 659,877(1μmol/L). In contrast, Ala NKA and NKA responses were only blocked by L 659,877. The oocytes injected with DH Poly(A) +RNA also responded to Sar SP and NKA with similar inward currents, which were selectively blocked by L 668,169 and L 659,877, respectively. These tachykinins induced responses had a potent desensitization. The present data indicate expression of NK 1 and NK 2 receptors in DRG neurons, suggesting that there may be tachykinin autoreceptors on the nociceptive primary afferent terminals.
