Anti-Proliferative Actions of T-Type Calcium Channel Inhibition in Thy1 Nephritis
Anti-Proliferative Actions of T-Type Calcium Channel Inhibition in Thy1 Nephritis
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DOI：
10.1016/j.ajpath.2013.04.029
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发表时间：
2013-08-01
影响因子：
6
通讯作者：
Hendry, Bruce M.
中科院分区：
文献类型：
--
作者：
Cove-Smith, Andrea;Mulgrew, Christopher J.;Hendry, Bruce M.
系膜细胞（MC）的异常增殖是进行性肾小球疾病的一个重要发现。 TH1177 是一种小分子，已被证明可以抑制低压激活的 T 型 Ca2+ 通道 (TCC)。目前的研究调查了 TH1177 对体外和体内 MC 增殖的影响。使用微量培养四唑测定并通过测量溴脱氧尿苷掺入来研究 Ca2+ 通道抑制对原代大鼠体外 MC 增殖的影响。在体内，用 TH1177 或媒介物治疗患有 Thy1 肾炎的大鼠。以盲法确定肾小球损伤和平均肾小球细胞数。还进行了 Ki-67 和磷酸化 ERK 的免疫染色。使用定量实时 PCR 研究 TCC 亚型在健康和患病组织中的表达。 TCC 阻断导致体外大鼠 MC 增殖显着减少，而 L 型抑制则没有效果。与安慰剂相比，用 TH1177 治疗 Thy1 肾炎可显着减少肾小球损伤 (P < 0.005)，并使肾小球细胞数量减少 49% (P < 0.005)。 TH1177 还将每个肾小球的 Ki-67 阳性和 pERK 阳性细胞减少了 52%（分别为 P < 0.01 和 P < 0.005）。这些结果表明，TH1177 在体外和体内抑制 MC 增殖，支持了以下假设：TCC 抑制可能是研究和改变 MC 对损伤的增殖反应的有用策略。
Aberrant proliferation of mesangial cells (MCs) is a key finding in progressive glomerular disease. TH1177 is a small molecule that has been shown to inhibit low-voltage activated T-type Ca2+ channels (TCCs). The current study investigates the effect of TH1177 on MC proliferation in vitro and in vivo. The effect of Ca2+ channel inhibition on primary rat MC proliferation in vitro was studied using the microculture tetrazolium assay and by measuring bromodeoxyuridine incorporation. In vivo, rats with Thy1 nephritis were treated with TH1177 or vehicle. Glomerular injury and average glomerular cell number were determined in a blinded fashion. Immunostaining for Ki-67 and phosphorylated ERK were also performed. The expression of TCC isoforms in healthy and diseased tissue was investigated using quantitative real-time PCR. TCC blockade caused a significant reduction in rat MC proliferation in vitro, whereas L-type inhibition had no effect. Treatment of Thy1 nephritis with TH1177 significantly reduced glomerular injury (P < 0.005) and caused a 49% reduction in glomerular cell number (P < 0.005) compared to the placebo. TH1177 also reduced Ki-67-positive and pERK-positive cells per glomerulus by 52% (P < 0.01 and P < 0.005, respectively). These results demonstrate that TH1177 inhibits MC proliferation in vitro and in vivo, supporting the hypothesis that TCC inhibition may be a useful strategy for studying and modifying MC proliferative responses to injury.