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首页> 美国卫生研究院文献>World Journal of Gastroenterology >Expression of gonadotropin-releasing hormone receptor and effect of gonadotropin-releasing hormone analogue on proliferation of cultured gastric smooth muscle cells of rats
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Expression of gonadotropin-releasing hormone receptor and effect of gonadotropin-releasing hormone analogue on proliferation of cultured gastric smooth muscle cells of rats

机译：促性腺激素释放激素受体的表达及促性腺激素释放激素类似物对大鼠胃平滑肌细胞增殖的影响

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AIM: To investigate the expression of gonadotropin-releasing hormone (GnRH) receptor and the effects of GnRH analog (alarelin) on proliferation of cultured gastric smooth muscle cells (GSMC) of rats.METHODS: Immunohistochemical ABC methods and in situ hybridization methods were used to dectect protein and mRNA expression of GnRH receptor in GSMC, respectively. Techniques of cell culture, OD value of MTT test, measure of 3H-TdR incorporation, average fluorescent values of proliferating cell nuclear antigen (PCNA) and flow cytometric DNA analysis were used in the experiment.RESULTS: The cultured GSMC of rats showed immunoreactivity for GnRH receptor; positive staining was located in cytoplasm. GnRH receptor mRNA hybridized signals were also detected in cytoplasm. When alarelin (10-9, 10-7, 10-5 mol/L) was administered into the medium and incubated for 24 h, OD value of MTT, 3H-TdR incorporation and average fluorescent values of PCNA all decreased significantly as compared with the control group (P < 0.05). The maximum inhibitory effect on cell proliferation was achieved a concentration of 10-5 mol/L and it acted in a dose-dependent manner. Flow cytometric DNA analysis revealed that alarelin could significantly enhance ratio of G1 phase and decrease ratio of S phase of GSMC of rats (P < 0.05).The maximum inhibitory effect on ratio of S phase was at the concentration of 10-5 mol/L and also acted in a dose-dependent manner.CONCLUSION: Our data suggest that GnRH receptor can be expressed by GSMC of rats. GnRH analogue can directly inhibit proliferation and DNA synthesis of rat GSMC through GnRH receptors.
机译：目的：探讨促性腺激素释放激素（GnRH）受体的表达以及GnRH类似物（阿拉瑞林）对大鼠培养的胃平滑肌细胞（GSMC）增殖的影响。方法：采用免疫组织化学ABC法和原位杂交法分别检测GSMC中GnRH受体的蛋白和mRNA表达。实验中采用细胞培养技术，MTT法OD值， 3 H-TdR掺入法，增殖细胞核抗原（PCNA）平均荧光值和流式细胞术DNA分析。培养的大鼠GSMC对GnRH受体具有免疫反应性。阳性染色位于细胞质中。在细胞质中也检测到GnRH受体mRNA杂交信号。当将阿拉瑞林（10 -9 ，10 -7 ，10 -5 mol / L）注入培养基中并孵育24小时时，与对照组相比，MTT的OD值， 3 H-TdR的掺入和PCNA的平均荧光值均明显降低（P <0.05）。浓度为10 -5 mol / L时，对细胞增殖的抑制作用最大，且呈剂量依赖性。流式细胞仪分析表明，阿拉瑞林能显着提高大鼠GSMC的G1期比例和降低S期比例（P <0.05）。对S期比例的最大抑制作用是在10 -5浓度下 mol / L，且也呈剂量依赖性。结论：我们的数据提示大鼠GSMC可以表达GnRH受体。 GnRH类似物可以通过GnRH受体直接抑制大鼠GSMC的增殖和DNA合成。

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