Peng He

Third Military Medical University, Ch’ung-ch’ing-shih, Chongqing Shi, China

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Publications (4)9.29 Total impact

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    ABSTRACT: OBJECTIVE: To investigate how the sodium/calcium exchanger subtype 3 (NCX3) and its reverse mode contribute to the function of interstitial cells of Cajal (ICCs) from the rat bladder. METHODS: The study used 20 female Wistar rats. We observed the expression of the NCX3 expression in the bladder using reverse transcriptase-polymerase chain reaction and Western blotting. The NCX3 in ICCs was also confirmed by double-labeled fluorescence. NCX3 functions in reverse mode of ICCs were observed using confocal microscopy with preload fluo-3AM, and its currents were evaluated using the whole-cell patch clamp technique, with or without the NCX3 inhibitor KB-R7943 (5 and 30μM), with an afterward identification of ICCs using single-cell polymerase chain reaction. RESULTS: NCX3 was confirmed in rat bladder ICCs. The time required for the intracellular calcium concentration [Ca(2+)]i of NCX3 was enhanced by KB-R7943 (5μM, P ≤.01). Moreover, KB-R7943 (5 and 30μM) significantly decreased the currents generated by the reverse mode of NCX3 from the ICCs (P <.05). CONCLUSION: NCX3 is expressed in rat bladder ICCs. The reverse mode of NCX3 can generate [Ca(2+)]i of the bladder ICCs.
    Urology 05/2013; · 2.42 Impact Factor
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    ABSTRACT: OBJECTIVE: To investigate the expression and function of T-type calcium channels in the interstitial cells of Cajal in rat bladders. METHODS: Bladders were harvested from Sprague-Dawley rats. The expression of T-type calcium channels subtypes (α1G, α1H, and α1I) in interstitial cells of Cajal were identified by double-labeled immunofluorescence analysis and reverse transcription-polymerase chain reaction analysis in whole mount preparations of rat bladders. The function of T-type calcium channels in freshly isolated interstitial cells of Cajal was assessed by detecting the changes of intracellular calcium ([Ca(2+)](i)) with preloading fluo-3 AM, and by evaluating the changes of the phasic contractions of rat bladder strips after treating with mibefradil and glivec. RESULTS: Three T-type calcium channels subtypes, α1G, α1H, and α1I, colocalized with c-kit in bladder interstitial cells of Cajal by double-labeled immunofluorescence analysis, and this was confirmed using reverse transcription-polymerase chain reaction. The T-type calcium channels selective blocker, mibefradil (1 μM), significantly decreased the intracellular calcium concentration ([Ca(2+)](i)) in isolated interstitial cells of Cajal (P < .01) and inhibited the spontaneous phasic contraction of bladder strips (P < .01). Moreover, the c-kit receptor blocker, glivec, significantly decreased the [Ca(2+)](i) of interstitial cells of Cajal further (P < .01) and the spontaneous phasic contraction of bladder strips. CONCLUSION: T-type calcium channel subtypes were confirmed to colocalize in interstitial cells of Cajal in rats bladders, which might participate in the spontaneous activity of interstitial cells of Cajal and phasic contractions of bladder strips by modulating [Ca(2+)](i) in interstitial cells of Cajal.
    Urology 09/2012; · 2.42 Impact Factor
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    ABSTRACT: To investigate the distribution and effects of hyperpolarization-activated cyclic nucleotide-gated (HCN) channel and its isoforms in bladder, especially in bladder interstitial cells of Cajal (ICC). Four HCN isoforms were detected in bladder tissue from rats using reverse transcription-polymerase chain reaction and Western blotting. The HCN1 subtype was observed in bladder ICCs by double-labeled fluorescence. The effect of the HCN blocker, ZD7288, was investigated using the bladder smooth muscle strip test. HCN1-4 isoforms were all identified in bladder ICCs using reverse transcription-polymerase chain reaction and Western blotting. Based on our semiquantitative analysis, HCN1 was found to be the most prominent isoform. The expression of HCN1 was confirmed in bladder ICCs by double-labeled fluorescence through colabeling of HCN1 and kit (CD117). ZD7288 significantly decreased the bladder excitation. All 4 HCN channel isoforms exist in the bladder, and they affect the bladder excitation, presumably via bladder ICCs.
    Urology 03/2012; 79(6):1411.e7-13. · 2.42 Impact Factor
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    ABSTRACT: To investigate the effects of imatinib (Glivec) on the urinary bladder contraction and excitation induced by neurostimulation, therefore to clarify the relationships between the bladder interstitial cell of Cajal (ICC) and the neural signals. In in vivo experiments, pelvic nerves of rats were stimulated by square-wave pulses. The contractile response was recorded before and 40 min after the administration of medications (atropine, Glivec, and ketotifen). In in vitro experiments, the bladder contractile response induced by acetylcholine with or without Glivec was evaluated. The space relationship between ICC and neural fibers were observed with double-labeled fluorescence using primary antibodies (anti-c-kit and anti-vesicular acetylcholine transferase) and secondary fluorescent antibodies (Alexa 488 and Alexa 594; Molecular Probes, Eugene, OR). Atropine and Glivec could significantly inhibit the bladder contractile response induced by the electrical stimulation in a dose-dependent manner, while ketotifen did not obviously affect bladder contractile response. In in vitro experiments, Glivec did not affect acetylcholine-induced bladder contractile response. The location of ICC in close proximity to cholinergic nerve fibers was confirmed by double-labeled fluorescence. Bladder ICC play an important role as intermediaries in the transmission of cholinergic signals from nerve to smooth muscle cells.
    Journal of Surgical Research 08/2011; 171(2):e193-9. · 2.02 Impact Factor