大麻二酚抑制创伤性脑损伤大鼠睾丸凋亡相关蛋白的表达  

Cannabidiol inhibits the express of apoptosis-related proteins in the testis of rats with traumatic brain injury

作　　者：李佳丽 凌腾晗 曹艳 尹爱平 李恒希 杨莉[2] 吴海鹰[2] 李坪[1] Li Jiali;Ling Tenghan;Cao Yan;Yin Aiping;Li Hengxi;Yang Li;Wu Haiying;Li Ping(Dept of Human Anatomy and Histology&Embryology,Kunming Medical University,Kunming 650500;Dept of Emergency,The First Affiliated Hospital of Kunming Medical University,Kunming 650032)

机构地区：[1]昆明医科大学人体解剖学与组织胚胎学系,昆明650500 [2]昆明医科大学第一附属医院急诊科,昆明650032

出　　处：《安徽医科大学学报》2024年第7期1128-1133,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82060241、82260387);云南省科技厅计划项目(编号:202101AY070001-002);云南省医疗机构科研项目(编号:202403AC100010);昆明医科大学研究生创新基金(编号:2023S133)。

摘　　要：目的研究创伤性脑损伤(TBI)后大鼠睾丸的损伤情况,并分析大麻二酚(CBD)对TBI引起的睾丸损伤的干预作用。方法将18只SD大鼠随机分为3组:假手术组(Sham组)、模型组(TBI组)和治疗组(TBI+CBD组)。采用HE染色法观察大鼠睾丸组织病理学变化,采用ELISA法检测大鼠血清中的睾酮水平。采用TUNEL染色法观察细胞凋亡,同时采用免疫荧光染色、Western blot和RT-qPCR法检测Bax、Bcl-2、Cleaved Caspase-3和肿瘤坏死因子-α(TNF-α)蛋白及mRNA表达。结果HE结果显示,与Sham组比较,TBI组大鼠睾丸出现病理改变。ELISA检测表明,与Sham组比较,TBI组的睾酮水平下降,但差异无统计学意义(P>0.05)。免疫荧光结果显示,与Sham组比较,TBI组大鼠睾丸Bax荧光表达强度升高(P<0.01);而CBD干预后Bax荧光强度下降且Bcl-2荧光强度升高(P<0.01)。Western blot结果表明,大鼠经CBD治疗后降低了睾丸凋亡相关蛋白(Bax和Cleaved Caspase-3,均P<0.05)和炎症相关蛋白(TNF-α,P<0.01)的蛋白水平,升高了抗凋亡蛋白Bcl-2的蛋白水平(P<0.05)。RT-qPCR结果趋势近似,与TBI组比较,CBD干预后Bax(P<0.05)和TNF-α(P<0.01)的mRNA表达下降,而Bcl-2的mRNA表达升高(P<0.05)。结论TBI导致睾丸损伤,CBD能够减轻TBI大鼠睾丸组织的凋亡和炎症反应。Objective To investigate testicular damage in traumatic brain injury(TBI)rats and to analyze the interventional effects of cannabidiol(CBD)on TBI-induced testicular damage.Methods 18 Sprague Dawley(SD)rats were randomly divided into three groups:the sham operation group(Sham group),model group(TBI group)and treatment group(TBI+CBD group).HE staining was used to observe the testicular histopathological changes in the rat testis.ELISA was used to detect testosterone level in rat serum.TUNEL staining was utilized to observe apoptosis,while immunofluorescence staining,Western blot and RT-qPCR were employed to evaluate Bax,Bcl-2,Cleaved Caspase-3 and TNF-αprotein and mRNA expression.Results HE staining showed pathological changes in the testes of TBI rats compared with those in the Sham group.ELISA assay showed a decrease in testosterone levels in the TBI group compared to the Sham group,but there was no significant difference(P>0.05).Immunofluorescence results showed that the intensity of Bax fluorescence expression increased in the TBI group compared with the Sham group(P<0.01),whereas the intensity of Bax fluorescence decreased and the intensity of Bcl-2 fluorescence increased in the rats after the CBD intervention(P<0.01).Western Blot results showed that CBD treatment in rats decreased the protein levels of testicular apoptosis-related proteins(Bax and Cleaved Caspase-3,all P<0.05)and inflammation-related proteins(TNF-α,P<0.01),and increased the protein level of the anti-apoptotic protein,Bcl-2(P<0.05).The trend of RT-qPCR results was similar,with mRNA expression of Bax(P<0.05)and TNF-α(P<0.01)decreased and mRNA expression of Bcl-2 increased after CBD intervention compared with the TBI group(P<0.05).Conclusion TBI induces testis injury,and CBD treatment effectively repairs apoptosis and inflammatory in testicular tissue of TBI rats.

关 键 词：大麻二酚 创伤性脑损伤 凋亡 炎症 大鼠 睾丸 

分 类 号：R742[医药卫生—神经病学与精神病学]