基于CD4~+CD25~+调节性T淋巴细胞特性探讨HBV携带者辨证论治的临床研究
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摘要
目的本课题以中医理论为指导,在国内外医学研究背景和导师临床经验的指导下,探讨慢性乙型肝炎中医辨证分型及HBeAg(+)与HBeAg(一)携带者外周血中CD4'CD25'调节性T细胞的变化及其意义,通过观察CD4+CD25'调节性T淋巴细胞的变化探讨中医药对HBV携带者进行干预后的免疫学特性。
方法收集2010年8月至2011年12月东南大学附属第二医院中西医结合肝病科门诊HBV携带者200例,进行中医证型及证候因素辨证；选取HBV携带者中肝郁脾虚型69例,均符合该病的诊断标准和纳入标准,分为空白对照组43例,其中HBeAg(+)组16例,HBeAg(一)组27例,苦参素组16例,当归补血汤加减组10例,另选取健康对照组25例。以HBeAg(+)组、HBeAg(一)组与健康对照组比较,得出HBV携带者免疫水平；再将HBV携带者与经过苦参素、当归补血汤加减的治疗组进行比较,观察中药对机体的免疫调节作用。检测各组的外周血CD3+T淋巴细胞所占比例、CD4+和CD8+T淋巴细胞所占比例及其比值、NK.NKT淋巴细胞所占比例及CD4+CD25'调节性T淋巴细胞所占比例。
结果HBeAg(+)与HBeAg(-)慢性乙型肝炎携带者在证型分布上均表现为以肝郁脾虚型为主,湿热中阻型、瘀血阻络型、肝肾阴虚型散在出现,均无脾肾阳虚型证候特点；HBeAg(+)组表现以湿证、热证为主,血瘀证次之,伴有不同程度的虚证；HBeAg(-)组以虚证为主,表现为阴虚,血虚,并伴有血瘀证候,湿热证候不显。HBeAg(+)组热证多于与HBeAg(-)组；而虚证少于与HBeAg(-)组,差异有统计学意义(P<0.05).HBeAg(+)、HBeAg(-)携带者与健康对照组相比,CD3+T淋巴细胞所占比例、CD4+和CD8+T淋巴细胞所占比例、NK.NKT淋巴细胞所占比例均无统计学意义,CD4+CD25+Tregs比例(%)分别为：4.82±3.49,2.15±2.36,2.02±0.82；HBeAg(+)组与健康对照组比较有统计学意义(p<0.05),HBeAg(-)组与健康对照组无统计学意义；苦参素组和当归补血汤组与HBeAg(+)组比较,外周血CD3+T淋巴细胞、CD4+与CD8+T淋巴细胞所占比例及比值差异无统计学意义,CD4'CD25'Treg比例(%)分别为：4.82±3.49,1.13±1.46,1.48±0.94：差异有统计学意义；与HBeAg(-)组相比,外周血CD3+T淋巴细胞、CD4+与CD8+T淋巴细胞所占比例及比值差异无统计学意义,CD4+CD25+Treg比例(%)分别为：2.15±2.36,1.13±1.46,1.48±0.94,差异无统计学意义；
结论(1)HBV携带者中医证型分布规律以肝郁脾虚型为主；(2)在证候因素上表现为HBeAg(+)组湿热证多于HBeAg(-)组;而虚证少于HBeAg(-)组;(3)HBeAg(+)、HBeAg(-)携带者相比于正常对照组,CD4+CD25+Tregs所占比例均有升高,提示处于不同的免疫状态；(4)中药苦参素与当归补血汤加减可下调慢性乙型肝炎携带者的CD4+CD25+Tregs水平,治疗组优于对照组,提示通过下调CD4+CD25+Tregs水平影响机体对乙肝病毒的免疫应答。
Objective With the guidance of the theroy of TCM,on the ground of in tutor formerly clinical study and the guidance of the professor, to investigate the distribution differences of syndrome in HBV and the Changes and Value of the proportion of CD4+CD25+regulatory T cells (Tregs) in HBeAg(+) and HBeAg(-) chronic asymptomtic hepatitis B virus carriers(ASC),and to explore the immune state after the treatment of oxymatrine and Chinese Angelica Decoction for Tonifying the Blood.
Methods200patients with HBV, collected from the second hospital of Nanjing, are deversed into HBeAg(+) group (100patients) and HBeAg(-) group (100patients). Investatigating the characteristics and distribution of syndrome types by syndrome differentiation of Traditional Chinese Medicine.Patients with HBeAg(+) HBV carriers (n=16),HBeAg(-)(n=27),treatment with oxymatrine(n=16),treatment with Chinese Angelica Decoction for Tonifying the Blood(n=10) and normol subjects(n=25) confirmed to the accepted standard are recruited in this study. First,Compared oxymatrine group and Chinese Angelica Decoction for Tonifying the Blood group to ASC,to observe the effect of Chinese medicine to the immune state of ASC. Detect the quantification of peripheral CD3+,CD4+,CD8+T lymphocyte subgroup,CD4+/CD8+,NK,NKT and CD4+CD25+Tregs.
Results The main proportion of ASC was syndrome of liver depression with spleen insufficiency, the rest were showed syndrome of damp-heat obstruction,syndrome of live-kidney yin deficency,syndrome of stagnant-blood obstructing collaterals,and there is no patient classified into syndrome of spleen-kidney yang deficiency. People in HBeAg(+) group shows a certain degree syndrome of dampness and heat,secondly is syndrome of blood stasis,accompanied by syndrome of deficiency.People in HBeAg(+) group mainly shows syndrome of yin and blood deficiency, accompanied by syndrome of blood stasis. The proportion of CD3,CD4+, CD8+T lymphocyte subgroup has no significant difference among healthy controls, HBeAg(+) and HBeAg(-) ASC. The proportion of CD4+CD25+Trcgs as follows(%):4.82±3.49,2.15±2.36,2.02±0.82;Contrast to healthy controls, The proportion of CD4+CD25+Tregs in HBeAg(+) patients is elevated(p<0.05),but there is no significant difference between healthy controls and HBeAg(-) patients. Compared oxymatrine group and Chinese Angelica Decoction for Tonifying the Blood group to HBeAg(+), The proportion of CD3+,CD4+,CD8+,CD4+/CD8+T lymphocyte subgroup has no significant difference,The proportion of CD4+CD25+Tregs as follows(%):4.82±3.49,1.13±1.46,1.48±0.94; Contrast to HBeAg(+) ASC,the proportion of CD3+,CD4+,CD8+T lymphocyte subgroup of oxymatrine and Chinese Angelica Decoction for Tonifying the Blood has no significant difference, but the proportion of CD4+CD25+Tregs is descent(p<0.05).Contrast to HBeAg(-) ASC,The proportion of CD3+,CD4+,CD8+,CD4+/CD8+, CD4+CD25+T lymphocyte subgroup has no significant difference,and the proportion of CD4+CD25+Tregs as follows(%):2.15±2.36,1.13±1.46,1.48±0.94.
Conclusion (1)The main proportion of ASC was syndrome of liver depression with spleen insufficiency.(2)Contrast to HBeAg(-) group,People in HBeAg(+) shows more percentage of syndrome of dampness and heat,but less percentage of syndrome of deficiency.(3)The proportion of CD4+CD25+Tregs in both HBeAg(+) and HBeAg(-)ASC is higher than health controls, showing that they are in different immune state.(4).oxymatrine and Chinese Angelica Decoction for Tonifying the Blood influence the immune response to HBV by downregulating the proportion of CD4+CD25+Tregs in ASC.
方法收集2010年8月至2011年12月东南大学附属第二医院中西医结合肝病科门诊HBV携带者200例,进行中医证型及证候因素辨证；选取HBV携带者中肝郁脾虚型69例,均符合该病的诊断标准和纳入标准,分为空白对照组43例,其中HBeAg(+)组16例,HBeAg(一)组27例,苦参素组16例,当归补血汤加减组10例,另选取健康对照组25例。以HBeAg(+)组、HBeAg(一)组与健康对照组比较,得出HBV携带者免疫水平；再将HBV携带者与经过苦参素、当归补血汤加减的治疗组进行比较,观察中药对机体的免疫调节作用。检测各组的外周血CD3+T淋巴细胞所占比例、CD4+和CD8+T淋巴细胞所占比例及其比值、NK.NKT淋巴细胞所占比例及CD4+CD25'调节性T淋巴细胞所占比例。
结果HBeAg(+)与HBeAg(-)慢性乙型肝炎携带者在证型分布上均表现为以肝郁脾虚型为主,湿热中阻型、瘀血阻络型、肝肾阴虚型散在出现,均无脾肾阳虚型证候特点；HBeAg(+)组表现以湿证、热证为主,血瘀证次之,伴有不同程度的虚证；HBeAg(-)组以虚证为主,表现为阴虚,血虚,并伴有血瘀证候,湿热证候不显。HBeAg(+)组热证多于与HBeAg(-)组；而虚证少于与HBeAg(-)组,差异有统计学意义(P<0.05).HBeAg(+)、HBeAg(-)携带者与健康对照组相比,CD3+T淋巴细胞所占比例、CD4+和CD8+T淋巴细胞所占比例、NK.NKT淋巴细胞所占比例均无统计学意义,CD4+CD25+Tregs比例(%)分别为：4.82±3.49,2.15±2.36,2.02±0.82；HBeAg(+)组与健康对照组比较有统计学意义(p<0.05),HBeAg(-)组与健康对照组无统计学意义；苦参素组和当归补血汤组与HBeAg(+)组比较,外周血CD3+T淋巴细胞、CD4+与CD8+T淋巴细胞所占比例及比值差异无统计学意义,CD4'CD25'Treg比例(%)分别为：4.82±3.49,1.13±1.46,1.48±0.94：差异有统计学意义；与HBeAg(-)组相比,外周血CD3+T淋巴细胞、CD4+与CD8+T淋巴细胞所占比例及比值差异无统计学意义,CD4+CD25+Treg比例(%)分别为：2.15±2.36,1.13±1.46,1.48±0.94,差异无统计学意义；
结论(1)HBV携带者中医证型分布规律以肝郁脾虚型为主；(2)在证候因素上表现为HBeAg(+)组湿热证多于HBeAg(-)组;而虚证少于HBeAg(-)组;(3)HBeAg(+)、HBeAg(-)携带者相比于正常对照组,CD4+CD25+Tregs所占比例均有升高,提示处于不同的免疫状态；(4)中药苦参素与当归补血汤加减可下调慢性乙型肝炎携带者的CD4+CD25+Tregs水平,治疗组优于对照组,提示通过下调CD4+CD25+Tregs水平影响机体对乙肝病毒的免疫应答。
Objective With the guidance of the theroy of TCM,on the ground of in tutor formerly clinical study and the guidance of the professor, to investigate the distribution differences of syndrome in HBV and the Changes and Value of the proportion of CD4+CD25+regulatory T cells (Tregs) in HBeAg(+) and HBeAg(-) chronic asymptomtic hepatitis B virus carriers(ASC),and to explore the immune state after the treatment of oxymatrine and Chinese Angelica Decoction for Tonifying the Blood.
Methods200patients with HBV, collected from the second hospital of Nanjing, are deversed into HBeAg(+) group (100patients) and HBeAg(-) group (100patients). Investatigating the characteristics and distribution of syndrome types by syndrome differentiation of Traditional Chinese Medicine.Patients with HBeAg(+) HBV carriers (n=16),HBeAg(-)(n=27),treatment with oxymatrine(n=16),treatment with Chinese Angelica Decoction for Tonifying the Blood(n=10) and normol subjects(n=25) confirmed to the accepted standard are recruited in this study. First,Compared oxymatrine group and Chinese Angelica Decoction for Tonifying the Blood group to ASC,to observe the effect of Chinese medicine to the immune state of ASC. Detect the quantification of peripheral CD3+,CD4+,CD8+T lymphocyte subgroup,CD4+/CD8+,NK,NKT and CD4+CD25+Tregs.
Results The main proportion of ASC was syndrome of liver depression with spleen insufficiency, the rest were showed syndrome of damp-heat obstruction,syndrome of live-kidney yin deficency,syndrome of stagnant-blood obstructing collaterals,and there is no patient classified into syndrome of spleen-kidney yang deficiency. People in HBeAg(+) group shows a certain degree syndrome of dampness and heat,secondly is syndrome of blood stasis,accompanied by syndrome of deficiency.People in HBeAg(+) group mainly shows syndrome of yin and blood deficiency, accompanied by syndrome of blood stasis. The proportion of CD3,CD4+, CD8+T lymphocyte subgroup has no significant difference among healthy controls, HBeAg(+) and HBeAg(-) ASC. The proportion of CD4+CD25+Trcgs as follows(%):4.82±3.49,2.15±2.36,2.02±0.82;Contrast to healthy controls, The proportion of CD4+CD25+Tregs in HBeAg(+) patients is elevated(p<0.05),but there is no significant difference between healthy controls and HBeAg(-) patients. Compared oxymatrine group and Chinese Angelica Decoction for Tonifying the Blood group to HBeAg(+), The proportion of CD3+,CD4+,CD8+,CD4+/CD8+T lymphocyte subgroup has no significant difference,The proportion of CD4+CD25+Tregs as follows(%):4.82±3.49,1.13±1.46,1.48±0.94; Contrast to HBeAg(+) ASC,the proportion of CD3+,CD4+,CD8+T lymphocyte subgroup of oxymatrine and Chinese Angelica Decoction for Tonifying the Blood has no significant difference, but the proportion of CD4+CD25+Tregs is descent(p<0.05).Contrast to HBeAg(-) ASC,The proportion of CD3+,CD4+,CD8+,CD4+/CD8+, CD4+CD25+T lymphocyte subgroup has no significant difference,and the proportion of CD4+CD25+Tregs as follows(%):2.15±2.36,1.13±1.46,1.48±0.94.
Conclusion (1)The main proportion of ASC was syndrome of liver depression with spleen insufficiency.(2)Contrast to HBeAg(-) group,People in HBeAg(+) shows more percentage of syndrome of dampness and heat,but less percentage of syndrome of deficiency.(3)The proportion of CD4+CD25+Tregs in both HBeAg(+) and HBeAg(-)ASC is higher than health controls, showing that they are in different immune state.(4).oxymatrine and Chinese Angelica Decoction for Tonifying the Blood influence the immune response to HBV by downregulating the proportion of CD4+CD25+Tregs in ASC.
引文
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