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目的:观察和分析大鼠睾丸局部短暂热应激对HSP70、HSP90、HSP105 mRNA表达的影响。方法:Wistar雄性大鼠随机分为5组:正常对照(N)组、热应激0天(H0)组、热应激5天(H5)组、热应激10天(H10)组、热应激15天(H15)组。N组睾丸局部22℃水浴20 min,其余各组均睾丸局部43℃水浴20 min。采用HE染色观察组织形态学变化,采用Real Time PCR的方法检测HSP70、HSP90、HSP105 mRNA的表达量。结果:HE染色镜下观察结果显示:与N组相比,H5组部分曲细精管萎缩,生精细胞明显消失,H10组、H15组大部分曲细精管萎缩,生精细胞大量消失。HE染色形态计量结果显示:与N组相比,H5组、H10组、H15组睾丸实质体积比明显减小(p<0.05),睾丸间质体积比明显增加(P<0.05)。RT-PCR结果显示:与N组相比,H0组HSP70 m RNA的表达H0组明显增高(p<0.05),H5组、H10组、H15组HSP90 m RNA的表达明显降低(P<0.05),H5组HSP105 m RNA的表达明显降低(P<0.05)。结论:HSP70、HSP90、HSP105 m RNA的表达在热应激后都会发生变化,变化的具体情况不尽相同,推测它们热应激后在生殖细胞凋亡过程中发挥不同的作用有关,并且和组织的损伤有密切的联系。
OBJECTIVE: To observe and analyze the effects of transient short-term heat stress on the mRNA expression of HSP70, HSP90 and HSP105 in rat testes. Methods: Wistar male rats were randomly divided into 5 groups: normal control group (N), heat stress 0 days (H0) group, heat stress 5 days (H5) group, heat stress 10 days Stress 15 days (H15) group. N group of testicular local 22 ℃ water bath 20 min, the rest of the testicular local 43 ℃ water bath 20 min. Histopathological changes were observed by HE staining. The expression of HSP70, HSP90 and HSP105 mRNA was detected by Real Time PCR. Results: HE staining showed that compared with group N, part of seminiferous tubules in group H5 were atrophied and spermatogenic cells were obviously disappeared. Most of seminiferous tubules in group H10 and group H15 were atrophied, and spermatogenic cells were largely disappeared. The results of HE staining showed that the volume ratio of testes in H5, H10 and H15 group was significantly decreased (p <0.05) and the interstitial volume was significantly increased (P <0.05) compared with N group. Compared with N group, the expression of HSP70mRNA in H0 group was significantly increased (P <0.05), while the expression of HSP90mRNA in H5 group, H10 group and H15 group was significantly lower than that in N group (P <0.05) The expression of HSP105 mRNA in H5 group was significantly decreased (P <0.05). CONCLUSION: The expression of HSP70, HSP90 and HSP105 m RNA changes after heat stress. The specific conditions of the changes are not the same. It is speculated that they play different roles in germ cell apoptosis after heat stress, The damage is closely linked.