目的探讨脑卒中患者应激性高血糖及导尿管相关尿路感染(CAUTI)易感性与大麻素受体1(CB1R)基因多态性的关系。方法选择武汉市汉口医院神经内科2020年5月-2021年5月因脑卒中住院接受治疗患者为研究对象,选择发生应激性高血糖和未发生应激...目的探讨脑卒中患者应激性高血糖及导尿管相关尿路感染(CAUTI)易感性与大麻素受体1(CB1R)基因多态性的关系。方法选择武汉市汉口医院神经内科2020年5月-2021年5月因脑卒中住院接受治疗患者为研究对象,选择发生应激性高血糖和未发生应激性高血糖患者各80例,分别纳入应激性高血糖组和非应激性高血糖组。进行聚合酶链式扩增反应扩增CB1R基因rs1049353位点目标序列并通过测序确定分型结果。统计患者性别、年龄、体质量指数、Charlson共病指数评分、脑卒中类型和CAUTI情况。统计患者入院后24 h内空腹血糖(FBG)和血糖不稳定指数(GLI)。统计所有应激性高血糖组入院后7、30 d神经功能缺损评分(NIHSS)和应激性高血糖组30 d预后情况。结果应激性高血糖组CAUTI发生率高于非应激性高血糖组(P<0.05);两组CB1R基因rs1049353位点基因型分布差异有统计学意义(P<0.05);应激性高血糖组CB1R基因rs1049353位点G等位基因和GG基因型频率低于非应激性高血糖组,AA基因型频率高于非应激性高血糖组(P<0.05);应激性高血糖组CB1R基因rs1049353位点携带AA基因型患者CAUTI发生率、入院当天FBG、GLI水平高于携带GG/GA型患者(P<0.05);应激性高血糖组CB1R基因rs1049353位点携带AA型和GG/GA型住院时间、30 d NIHSS评分、30 d存活情况比较差异无统计学意义,AA型7 d NIHSS评分高于GG/GA型(P<0.05)。结论CB1R基因rs1049353位点AA基因型增加脑卒中患者应激性高血糖和CAUTI风险,其机制还需进一步研究。展开更多
The rostral agranular insular cortex(RAIC)has been associated with pain modulation.Although the endogenous cannabinoid system(eCB)has been shown to regulate chronic pain,the roles of eCBs in the RAIC remain elusive un...The rostral agranular insular cortex(RAIC)has been associated with pain modulation.Although the endogenous cannabinoid system(eCB)has been shown to regulate chronic pain,the roles of eCBs in the RAIC remain elusive under the neuropathic pain state.Neuropathic pain was induced in C57BL/6 mice by common peroneal nerve(CPN)ligation.The roles of the eCB were tested in the RAIC of ligated CPN C57BL/6J mice,glutamatergic,or GABAergic neuron cannabinoid receptor 1(CB1R)knockdown mice with the whole-cell patch-clamp and pain behavioral methods.The E/I ratio(amplitude ratio between mEPSCs and mIPSCs)was significantly increased in layer V pyramidal neurons of the RAIC in CPN-ligated mice.Depolarization-induced suppression of inhibition but not depolarization-induced suppression of excitation in RAIC layer V pyramidal neurons were significantly increased in CPN-ligated mice.The analgesic effect of ACEA(a CB1R agonist)was alleviated along with bilateral dorsolateral funiculus lesions,with the administration of AM251(a CB1R antagonist),and in CB1R knockdown mice in GABAergic neurons,but not glutamatergic neurons of the RAIC.Our results suggest that CB1R activation reinforces the function of the descending pain inhibitory pathway via reducing the inhibition of glutamatergic layer V neurons by GABAergic neurons in the RAIC to induce an analgesic effect in neuropathic pain.展开更多
Many researchers employed mammalian expression system to artificially express cannabinoid receptors, but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports....Many researchers employed mammalian expression system to artificially express cannabinoid receptors, but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports. In present study, we demonstrated cannabinoid receptor protein was not able to be properly expressed with routine mammalian expression system. This inefficient expression was rescued by endowing an exogenous signal peptide ahead of cannabinoid receptor peptide. In addition, the artificially synthesized cannabinoid receptor was found to aggregate under routine sample denaturing temperatures (i.e.,≥95°C), forming a large molecular weight band when analyzed by immuno-blotting. Only denaturing temperatures ≤75°C yielded a clear band at the predicted molecular weight. Collectively, we showed that efficient mammalian expression of cannabinoid receptors need a signal peptide sequence, and described the requirement for a low sample denaturing temperature in immuno-blot analysis. These findings provide very useful information for efficient mammalian expression and immuno-blotting of membrane receptors.展开更多
文摘目的探讨脑卒中患者应激性高血糖及导尿管相关尿路感染(CAUTI)易感性与大麻素受体1(CB1R)基因多态性的关系。方法选择武汉市汉口医院神经内科2020年5月-2021年5月因脑卒中住院接受治疗患者为研究对象,选择发生应激性高血糖和未发生应激性高血糖患者各80例,分别纳入应激性高血糖组和非应激性高血糖组。进行聚合酶链式扩增反应扩增CB1R基因rs1049353位点目标序列并通过测序确定分型结果。统计患者性别、年龄、体质量指数、Charlson共病指数评分、脑卒中类型和CAUTI情况。统计患者入院后24 h内空腹血糖(FBG)和血糖不稳定指数(GLI)。统计所有应激性高血糖组入院后7、30 d神经功能缺损评分(NIHSS)和应激性高血糖组30 d预后情况。结果应激性高血糖组CAUTI发生率高于非应激性高血糖组(P<0.05);两组CB1R基因rs1049353位点基因型分布差异有统计学意义(P<0.05);应激性高血糖组CB1R基因rs1049353位点G等位基因和GG基因型频率低于非应激性高血糖组,AA基因型频率高于非应激性高血糖组(P<0.05);应激性高血糖组CB1R基因rs1049353位点携带AA基因型患者CAUTI发生率、入院当天FBG、GLI水平高于携带GG/GA型患者(P<0.05);应激性高血糖组CB1R基因rs1049353位点携带AA型和GG/GA型住院时间、30 d NIHSS评分、30 d存活情况比较差异无统计学意义,AA型7 d NIHSS评分高于GG/GA型(P<0.05)。结论CB1R基因rs1049353位点AA基因型增加脑卒中患者应激性高血糖和CAUTI风险,其机制还需进一步研究。
基金This work was supported by the National Natural Science Foundation of China(32271056,81671081,and 81701095)University Science and Technology Fund Planning Projects(2022XC002 and 2019XB006).
文摘The rostral agranular insular cortex(RAIC)has been associated with pain modulation.Although the endogenous cannabinoid system(eCB)has been shown to regulate chronic pain,the roles of eCBs in the RAIC remain elusive under the neuropathic pain state.Neuropathic pain was induced in C57BL/6 mice by common peroneal nerve(CPN)ligation.The roles of the eCB were tested in the RAIC of ligated CPN C57BL/6J mice,glutamatergic,or GABAergic neuron cannabinoid receptor 1(CB1R)knockdown mice with the whole-cell patch-clamp and pain behavioral methods.The E/I ratio(amplitude ratio between mEPSCs and mIPSCs)was significantly increased in layer V pyramidal neurons of the RAIC in CPN-ligated mice.Depolarization-induced suppression of inhibition but not depolarization-induced suppression of excitation in RAIC layer V pyramidal neurons were significantly increased in CPN-ligated mice.The analgesic effect of ACEA(a CB1R agonist)was alleviated along with bilateral dorsolateral funiculus lesions,with the administration of AM251(a CB1R antagonist),and in CB1R knockdown mice in GABAergic neurons,but not glutamatergic neurons of the RAIC.Our results suggest that CB1R activation reinforces the function of the descending pain inhibitory pathway via reducing the inhibition of glutamatergic layer V neurons by GABAergic neurons in the RAIC to induce an analgesic effect in neuropathic pain.
基金supported by a grant from Army Medical Research Program of China(No.08G168)
文摘Many researchers employed mammalian expression system to artificially express cannabinoid receptors, but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports. In present study, we demonstrated cannabinoid receptor protein was not able to be properly expressed with routine mammalian expression system. This inefficient expression was rescued by endowing an exogenous signal peptide ahead of cannabinoid receptor peptide. In addition, the artificially synthesized cannabinoid receptor was found to aggregate under routine sample denaturing temperatures (i.e.,≥95°C), forming a large molecular weight band when analyzed by immuno-blotting. Only denaturing temperatures ≤75°C yielded a clear band at the predicted molecular weight. Collectively, we showed that efficient mammalian expression of cannabinoid receptors need a signal peptide sequence, and described the requirement for a low sample denaturing temperature in immuno-blot analysis. These findings provide very useful information for efficient mammalian expression and immuno-blotting of membrane receptors.
