陽(yáng)虛質(zhì)慢性失眠患者NK細(xì)胞和T細(xì)胞的特征研究
發(fā)布時(shí)間：2019-05-23 15:49
【摘要】：目的:通過(guò)探索陽(yáng)虛質(zhì)慢性失眠患者的淋巴細(xì)胞亞群、NK細(xì)胞亞群及所含顆粒酶、穿孔素、NK細(xì)胞殺傷功能及分泌功能、Th1、Th2、Th17細(xì)胞亞群變化,從NK細(xì)胞和T細(xì)胞的角度分析陽(yáng)虛質(zhì)慢性失眠患者的免疫紊亂狀態(tài),為陽(yáng)虛質(zhì)慢性失眠的微觀化辨證提供理論依據(jù)。方法:本研究為橫斷面研究,納入心理睡眠門診失眠病人及健康體檢人群,根據(jù)失眠病史的有無(wú)分為慢性失眠組和正常對(duì)照組,并進(jìn)一步將慢性失眠組分別分為陽(yáng)虛質(zhì)組、非陽(yáng)虛質(zhì)組兩個(gè)亞組及陽(yáng)虛質(zhì)組、非陽(yáng)虛質(zhì)組偏頗體質(zhì)組、平和質(zhì)組三個(gè)亞組,收集一般資料、中醫(yī)體質(zhì)調(diào)查問(wèn)卷、匹茲堡睡眠量表,使用流式細(xì)胞術(shù)檢測(cè)淋巴細(xì)胞亞群檢、NK細(xì)胞殺傷功能、NK細(xì)胞亞群及所含顆粒酶、穿孔素、Th1/Th2/Th17細(xì)胞亞群,使用Elisa法檢測(cè)NK細(xì)胞分泌的INFγ、TNF a水平。對(duì)研究結(jié)果進(jìn)行統(tǒng)計(jì)分析,比較慢性失眠組與正常對(duì)照組間,慢性失眠組兩個(gè)亞組間及三個(gè)亞組間的一般資料、PSQI量表及上述檢測(cè)指標(biāo)的差異,并分析陽(yáng)虛程度與失眠程度及上述檢測(cè)指標(biāo)的相關(guān)性。結(jié)果:1.淋巴細(xì)胞亞群:慢性失眠組(n=59)的淋巴細(xì)胞總數(shù)、NK細(xì)胞數(shù)、總T淋巴細(xì)胞數(shù)、抑制性/細(xì)胞毒性T細(xì)胞數(shù)、總B淋巴細(xì)胞數(shù)較正常對(duì)照組(n=30)下降。慢性失眠組內(nèi),在陽(yáng)虛質(zhì)組與非陽(yáng)虛質(zhì)組兩組間比較中,陽(yáng)虛質(zhì)組的NK細(xì)胞數(shù)、NK細(xì)胞百分比較非陽(yáng)虛質(zhì)組下降,而總T淋巴細(xì)胞數(shù)、總T淋巴細(xì)胞百分比、輔助性T細(xì)胞數(shù)則較非陽(yáng)虛質(zhì)組上升。慢性失眠組內(nèi),在陽(yáng)虛質(zhì)組、非陽(yáng)虛質(zhì)偏頗體質(zhì)組、平和質(zhì)組三組間的比較中,陽(yáng)虛質(zhì)組的NK細(xì)胞百分比較平和質(zhì)組下降。慢性失眠組中總T淋巴細(xì)胞數(shù)(r=0.273)、總T淋巴細(xì)胞百分比(r=0.37)與陽(yáng)虛程度呈正相關(guān)關(guān)系,NK細(xì)胞百分比與陽(yáng)虛程度呈負(fù)相關(guān)關(guān)系(r=-0.273)。2.NK細(xì)胞亞群及所含顆粒酶、穿孔素:CD56+CD16-NK細(xì)胞亞群:慢性失眠組(n=32)的CD56+CD16-NK細(xì)胞亞群百分比較正常對(duì)照組(n=28)升高。慢性失眠組內(nèi),在陽(yáng)虛質(zhì)組與非陽(yáng)虛質(zhì)組兩組間的比較中,陽(yáng)虛質(zhì)組CD56+CD16-NK細(xì)胞亞群的顆粒酶百分比、穿孔素百分比、顆粒酶熒光強(qiáng)度均較非陽(yáng)虛質(zhì)組下降。慢性失眠組內(nèi),在陽(yáng)虛質(zhì)組、非陽(yáng)虛質(zhì)偏頗體質(zhì)組、平和體質(zhì)組三組間的比較中,陽(yáng)虛質(zhì)組的CD56+CD16-NK細(xì)胞亞群的顆粒酶百分比較平和質(zhì)組下降,而陽(yáng)虛質(zhì)組的CD56+CD16-NK細(xì)胞亞群的顆粒酶熒光強(qiáng)度較非陽(yáng)虛質(zhì)偏頗體質(zhì)組、平和質(zhì)組下降。CD56+CD16+NK細(xì)胞亞群:慢性失眠組(n=32)的CD56+CD16+NK細(xì)胞亞群百分比較正常對(duì)照組(n=28)下降,該亞群所含穿孔素百分比及穿孔素?zé)晒鈴?qiáng)度較正常對(duì)照組上升。慢性失眠組內(nèi),在陽(yáng)虛質(zhì)組與非陽(yáng)虛質(zhì)組兩組間的比較中,陽(yáng)虛質(zhì)組的CD56+CD16+NK細(xì)胞百分比、顆粒酶百分比、穿孔素百分比、穿孔素?zé)晒鈴?qiáng)度均較非陽(yáng)虛質(zhì)組下降。慢性失眠組內(nèi),在陽(yáng)虛質(zhì)組、非陽(yáng)虛質(zhì)偏頗體質(zhì)組、平和體質(zhì)組三組間的比較中,陽(yáng)虛質(zhì)組CD56+CD16+NK細(xì)胞亞群顆粒酶百分比較非陽(yáng)虛質(zhì)偏頗體質(zhì)組、平和質(zhì)組下降。陽(yáng)虛質(zhì)組CD56+CD16+NK細(xì)胞亞群穿孔素?zé)晒鈴?qiáng)度較平和質(zhì)組下降。CD56-CD16+NK細(xì)胞:慢性失眠組(n=32)的CD56-CD16+NK細(xì)胞的顆粒酶百分比較正常對(duì)照組(n=28)減少,而顆粒酶熒光強(qiáng)度、穿孔素?zé)晒鈴?qiáng)度均較正常對(duì)照組增加。慢性失眠組內(nèi),在陽(yáng)虛質(zhì)組、非陽(yáng)虛質(zhì)組兩組間的比較中,陽(yáng)虛質(zhì)組的CD56-CD16+NK細(xì)胞亞群的顆粒酶百分比、穿孔素百分比、穿孔素?zé)晒鈴?qiáng)度均較非陽(yáng)虛質(zhì)組下降。慢性失眠組內(nèi),在陽(yáng)虛質(zhì)組、非陽(yáng)虛質(zhì)偏頗體質(zhì)組、平和體質(zhì)組三組間的比較中,陽(yáng)虛質(zhì)組的CD56-CD16+NK細(xì)胞顆粒酶百分比、穿孔素百分比均較非陽(yáng)虛質(zhì)偏頗體質(zhì)組減少,而陽(yáng)虛質(zhì)組的CD56-CD16+NK細(xì)胞穿孔素?zé)晒鈴?qiáng)度則較平和質(zhì)組減少。慢性失眠組中,陽(yáng)虛程度與CD56+CD16+NK細(xì)胞亞群百分比(r=-0.446)、CD56+CD16+NK細(xì)胞亞群顆粒酶百分比(r=-0.647)、CD56+CD16+NK細(xì)胞亞群穿孔素?zé)晒饬?r=-0.465)、CD56+CD16-NK細(xì)胞亞群顆粒酶熒光量(r=-0.504)呈負(fù)相關(guān)關(guān)系。3.NK細(xì)胞殺傷功能及分泌功能:慢性失眠組(n=8)的NK細(xì)胞殺傷率、NK細(xì)胞分泌的TNFα、IFNy與正常對(duì)照組(n=7)無(wú)明顯差異。4.Th1、Th2、Th17 細(xì)胞亞群:慢性失眠組(n=58)的Th1/Th2比值較正常對(duì)照組(n=22)下降。在慢性失眠組內(nèi),在陽(yáng)虛質(zhì)組、非陽(yáng)虛質(zhì)組兩組間的比較中,陽(yáng)虛質(zhì)組的Th1細(xì)胞百分比、Th2細(xì)胞百分比、Th17細(xì)胞百分比均較非陽(yáng)虛質(zhì)組下降。在慢性失眠組內(nèi),在陽(yáng)虛質(zhì)組、非陽(yáng)虛質(zhì)偏頗體質(zhì)組、平和體質(zhì)組三組間的比較中,陽(yáng)虛質(zhì)組Th1細(xì)胞百分比、Th2細(xì)胞百分比均較非陽(yáng)虛質(zhì)偏頗體質(zhì)組下降,而陽(yáng)虛質(zhì)組Th17細(xì)胞百分比則較非陽(yáng)虛質(zhì)偏頗體質(zhì)組、平和質(zhì)組下降。慢性失眠組中,陽(yáng)虛程度與Th1細(xì)胞百分比(r=-0.452)、Th2細(xì)胞百分比(r=-0.39)、Th17細(xì)胞百分比(r=-0.387)細(xì)胞亞群呈負(fù)相關(guān)關(guān)系(P0.05)。結(jié)論:慢性失眠導(dǎo)致的免疫紊亂主要特征為NK細(xì)胞、T細(xì)胞數(shù)目的下降及相關(guān)功能亞群的偏移,其中的陽(yáng)虛質(zhì)慢性失眠還具有免疫激活能力下降的特征表現(xiàn),且陽(yáng)虛程度與上述指標(biāo)變化程度存在相關(guān)關(guān)系,提示陽(yáng)虛質(zhì)慢性失眠是慢性失眠中免疫功能較易受失眠影響的一種類型。
[Abstract]:Objective: To study the lymphocyte subsets, NK cell subsets and the changes of the secretory function, Th1, Th2 and Th17 subsets in the patients with chronic insomnia with yang-yang deficiency. The immune disorder status of the patients with yang-deficiency and chronic insomnia is analyzed from the angle of NK cells and T-cells, and the theoretical basis for the micro-differentiation of the chronic insomnia with yang-deficiency is provided. Methods: The study was a cross-sectional study, which was included in the patients with insomnia and the healthy physical examination of the patients with insomnia. According to the history of insomnia, the patients were divided into the chronic insomnia group and the normal control group, and the chronic insomnia group was divided into two groups of Yang-Yang, non-Yang, and Yang-Yang, respectively. The three sub-groups of the non-yang-yang mass group, the partial constitution group and the mild mass group, the general information, the physical constitution questionnaire of the traditional Chinese medicine and the Pittsburgh sleep scale were collected, and the lymphocyte subgroup detection, the NK cell killing function, the NK cell subpopulation and the particle-containing enzyme and the perforin were detected by flow cytometry. The subsets of Th1/ Th2/ Th17 cells and the level of TNF a, which were secreted by NK cells, were detected by the Elisa method. The general data, PSQI scale and the difference between the two sub-groups and the three sub-groups in the chronic insomnia group and the normal control group, the PSQI scale and the above-mentioned detection index were compared, and the correlation between the degree of yang-yang and the level of insomnia and the above-mentioned detection index was analyzed. Results:1. The lymphocyte subpopulation: the total number of lymphocytes, the number of NK cells, the total T-lymphocyte number, the inhibitory/ cytotoxic T-cell number and the total B-lymphocyte number of the chronic insomnia group (n = 59) decreased with the normal control group (n = 30). In the chronic insomnia group, the number of NK cells and the percentage of NK cells in the Yang-Yang mass group were lower than that of the non-Yang-Yang mass group, while the total number of T-lymphocytes, the percentage of total T-lymphocytes and the number of helper T-cells were higher than that of the non-Yang-Yang mass group. In the chronic insomnia group, the percentage of NK cells in the Yang-Yang mass group was lower than that in the mass group. The total number of T lymphocytes in the chronic insomnia group (r = 0.273), the total T-lymphocyte percentage (r = 0.37) was positively related to the degree of yang-yang, and the percentage of NK cells was negatively correlated with the degree of yang-yang (r =-0.273). The percentage of CD56 + CD16-NK cells in the chronic insomnia group (n = 32) was higher than that in the normal control group (n = 28). In the chronic insomnia group, the percentage of the particles of CD56 + CD16-NK cell subpopulation, the percentage of perforin and the fluorescence intensity of the granular enzyme were lower than that of the non-Yang-Yang mass group. In the chronic insomnia group, the percentage of the granular enzyme of CD56 + CD16-NK cell subpopulation of the Yang-Yang mass group was lower than that in the mass group. However, the fluorescence intensity of CD56 + CD16-NK cell subsets in the Yang-Yang mass group was lower than that of the non-yang-deficient group. The percentage of CD56 + CD16 + NK cells in the chronic insomnia group (n = 32) was lower than that in the normal control group (n = 28). In the chronic insomnia group, the percentage of CD56 + CD16 + NK cells, the percentage of granular enzyme, the percentage of perforin and the fluorescence intensity of the perforin were lower than that of the non-Yang-Yang mass group. In the chronic insomnia group, the percentage of the granular enzyme of CD56 + CD16 + NK cell subpopulation in the Yang-Yang mass group was lower than that of the non-yang-deficient group and the quality group decreased in the comparison among the three groups. The fluorescence intensity of CD56 + CD16 + NK cell subpopulation in the Yang-Yang mass group was lower than that in the mass group. CD56-CD16 + NK cells: the percentage of the particles of CD56-CD16 + NK cells in the chronic insomnia group (n = 32) was lower than that in the normal control group (n = 28), while the fluorescence intensity of the granular enzyme and the fluorescence intensity of the perforin were higher than that of the normal control group. In the chronic insomnia group, the percentage of the particles of the CD56-CD16 + NK cell subpopulation of the Yang-Yang mass group, the percentage of the perforin and the fluorescent intensity of the perforin were lower than that of the non-Yang-Yang mass group in the comparison among the two groups of the Yang-Yang mass group and the non-Yang-Yang mass group. In the chronic insomnia group, the percentage of CD56-CD16 + NK cell particles and the percentage of perforin in the Yang-Yang mass group were lower than that of the non-yang-deficient group. The fluorescence intensity of CD56-CD16 + NK cells in the Yang-Yang mass group was lower than that of the mild group. The percentage of CD56 + CD16 + NK cell subpopulation (r =-0.446), CD56 + CD16 + NK cell subpopulation particle enzyme percentage (r =-0.465), CD56 + CD16 + NK cell subpopulation particle enzyme fluorescence (r =-0.465), CD56 + CD16-NK cell subpopulation particle enzyme fluorescence (r =-0.504) were negatively correlated with the chronic insomnia group. The NK cell killing rate of the chronic insomnia group (n = 8), the TNF level of NK cell secretion, IFNy and the normal control group (n = 7) had no significant difference.4. The ratio of Th1/ Th2 and Th17 cells in chronic insomnia group (n = 58) was lower than that of normal control group (n = 22). In the chronic insomnia group, the percentage of Th1 cells, the percentage of Th2 cells and the percentage of Th17 cells in the Yang-Yang mass group were lower than that of the non-Yang-Yang mass group. in the chronic insomnia group, the percentage of the Th1 cells and the percentage of the Th2 cells in the Yang-Yang mass group were lower than that of the non-yang-deficient group, The percentage of Th17 cells in the Yang-Yang mass group was lower than that of the non-yang-deficient group. The percentage (r =-0.452), the percentage of Th2 cells (r =-0.39) and the percentage of Th17 cells (r =-0.387) in the chronic insomnia group were negatively correlated with the percentage of Th1 cells (r =-0.452), the percentage of Th2 cells (r =-0.39), and the percentage of Th17 cells (r =-0.387) (P0.05). Conclusion: The immune disorder caused by chronic insomnia is mainly characterized by the decrease of the number of NK cells and T cells and the shift of related functional subpopulations. It is suggested that chronic insomnia with yang-deficiency can be a type of immune function in chronic insomnia which is more likely to be affected by insomnia.
【學(xué)位授予單位】：廣州中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R256.23
本文編號(hào)：2484026
[Abstract]:Objective: To study the lymphocyte subsets, NK cell subsets and the changes of the secretory function, Th1, Th2 and Th17 subsets in the patients with chronic insomnia with yang-yang deficiency. The immune disorder status of the patients with yang-deficiency and chronic insomnia is analyzed from the angle of NK cells and T-cells, and the theoretical basis for the micro-differentiation of the chronic insomnia with yang-deficiency is provided. Methods: The study was a cross-sectional study, which was included in the patients with insomnia and the healthy physical examination of the patients with insomnia. According to the history of insomnia, the patients were divided into the chronic insomnia group and the normal control group, and the chronic insomnia group was divided into two groups of Yang-Yang, non-Yang, and Yang-Yang, respectively. The three sub-groups of the non-yang-yang mass group, the partial constitution group and the mild mass group, the general information, the physical constitution questionnaire of the traditional Chinese medicine and the Pittsburgh sleep scale were collected, and the lymphocyte subgroup detection, the NK cell killing function, the NK cell subpopulation and the particle-containing enzyme and the perforin were detected by flow cytometry. The subsets of Th1/ Th2/ Th17 cells and the level of TNF a, which were secreted by NK cells, were detected by the Elisa method. The general data, PSQI scale and the difference between the two sub-groups and the three sub-groups in the chronic insomnia group and the normal control group, the PSQI scale and the above-mentioned detection index were compared, and the correlation between the degree of yang-yang and the level of insomnia and the above-mentioned detection index was analyzed. Results:1. The lymphocyte subpopulation: the total number of lymphocytes, the number of NK cells, the total T-lymphocyte number, the inhibitory/ cytotoxic T-cell number and the total B-lymphocyte number of the chronic insomnia group (n = 59) decreased with the normal control group (n = 30). In the chronic insomnia group, the number of NK cells and the percentage of NK cells in the Yang-Yang mass group were lower than that of the non-Yang-Yang mass group, while the total number of T-lymphocytes, the percentage of total T-lymphocytes and the number of helper T-cells were higher than that of the non-Yang-Yang mass group. In the chronic insomnia group, the percentage of NK cells in the Yang-Yang mass group was lower than that in the mass group. The total number of T lymphocytes in the chronic insomnia group (r = 0.273), the total T-lymphocyte percentage (r = 0.37) was positively related to the degree of yang-yang, and the percentage of NK cells was negatively correlated with the degree of yang-yang (r =-0.273). The percentage of CD56 + CD16-NK cells in the chronic insomnia group (n = 32) was higher than that in the normal control group (n = 28). In the chronic insomnia group, the percentage of the particles of CD56 + CD16-NK cell subpopulation, the percentage of perforin and the fluorescence intensity of the granular enzyme were lower than that of the non-Yang-Yang mass group. In the chronic insomnia group, the percentage of the granular enzyme of CD56 + CD16-NK cell subpopulation of the Yang-Yang mass group was lower than that in the mass group. However, the fluorescence intensity of CD56 + CD16-NK cell subsets in the Yang-Yang mass group was lower than that of the non-yang-deficient group. The percentage of CD56 + CD16 + NK cells in the chronic insomnia group (n = 32) was lower than that in the normal control group (n = 28). In the chronic insomnia group, the percentage of CD56 + CD16 + NK cells, the percentage of granular enzyme, the percentage of perforin and the fluorescence intensity of the perforin were lower than that of the non-Yang-Yang mass group. In the chronic insomnia group, the percentage of the granular enzyme of CD56 + CD16 + NK cell subpopulation in the Yang-Yang mass group was lower than that of the non-yang-deficient group and the quality group decreased in the comparison among the three groups. The fluorescence intensity of CD56 + CD16 + NK cell subpopulation in the Yang-Yang mass group was lower than that in the mass group. CD56-CD16 + NK cells: the percentage of the particles of CD56-CD16 + NK cells in the chronic insomnia group (n = 32) was lower than that in the normal control group (n = 28), while the fluorescence intensity of the granular enzyme and the fluorescence intensity of the perforin were higher than that of the normal control group. In the chronic insomnia group, the percentage of the particles of the CD56-CD16 + NK cell subpopulation of the Yang-Yang mass group, the percentage of the perforin and the fluorescent intensity of the perforin were lower than that of the non-Yang-Yang mass group in the comparison among the two groups of the Yang-Yang mass group and the non-Yang-Yang mass group. In the chronic insomnia group, the percentage of CD56-CD16 + NK cell particles and the percentage of perforin in the Yang-Yang mass group were lower than that of the non-yang-deficient group. The fluorescence intensity of CD56-CD16 + NK cells in the Yang-Yang mass group was lower than that of the mild group. The percentage of CD56 + CD16 + NK cell subpopulation (r =-0.446), CD56 + CD16 + NK cell subpopulation particle enzyme percentage (r =-0.465), CD56 + CD16 + NK cell subpopulation particle enzyme fluorescence (r =-0.465), CD56 + CD16-NK cell subpopulation particle enzyme fluorescence (r =-0.504) were negatively correlated with the chronic insomnia group. The NK cell killing rate of the chronic insomnia group (n = 8), the TNF level of NK cell secretion, IFNy and the normal control group (n = 7) had no significant difference.4. The ratio of Th1/ Th2 and Th17 cells in chronic insomnia group (n = 58) was lower than that of normal control group (n = 22). In the chronic insomnia group, the percentage of Th1 cells, the percentage of Th2 cells and the percentage of Th17 cells in the Yang-Yang mass group were lower than that of the non-Yang-Yang mass group. in the chronic insomnia group, the percentage of the Th1 cells and the percentage of the Th2 cells in the Yang-Yang mass group were lower than that of the non-yang-deficient group, The percentage of Th17 cells in the Yang-Yang mass group was lower than that of the non-yang-deficient group. The percentage (r =-0.452), the percentage of Th2 cells (r =-0.39) and the percentage of Th17 cells (r =-0.387) in the chronic insomnia group were negatively correlated with the percentage of Th1 cells (r =-0.452), the percentage of Th2 cells (r =-0.39), and the percentage of Th17 cells (r =-0.387) (P0.05). Conclusion: The immune disorder caused by chronic insomnia is mainly characterized by the decrease of the number of NK cells and T cells and the shift of related functional subpopulations. It is suggested that chronic insomnia with yang-deficiency can be a type of immune function in chronic insomnia which is more likely to be affected by insomnia.
【學(xué)位授予單位】：廣州中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R256.23
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