温阳化浊通络方对系统性硬化病患者外周血Th17/Treg细胞平衡的影响

Effect of Wenyang Huazhuo Tongluo Recipe on Peripheral Blood Th17/Treg Cell Balance in Systemic Sclerosis Patients

目的观察温阳化浊通络方对系统性硬化病(systemic scierosis,SSc)阳气亏虚、浊邪阻络证患者外周血辅助性T细胞17(T helper 17 cells,Th17)/调节性T细胞(regulatory T cells,Treg)及其细胞因子IL-17、IL-10水平的影响。方法将82例SSc患者按随机数字表分为西医组(对照组)和中西医结合组(治疗组),每组各41例。两组均予甲氨蝶呤片、泼尼松片口服治疗,治疗组加温阳化浊通络方口服,两组均连续治疗6个月。另选择70名健康志愿者作为健康对照组(健康组)。采用流式细胞术检测外周血Th17和Treg细胞数量,采取双抗体夹心E LISA法检测血清IL-17、IL-10、血管性血友病因子(von Willebrand factor,vWF)、Ⅲ型前胶原氨基端原肽(aminoterminal propeptide of typeⅢprocollagen,PⅢNP)、Ⅰ型胶原羧基交联末肽(cross-linked carboxyterminal telopeptide of typeⅠcollagen,ⅠCTP)水平,采用Pearson相关分析观察Th17/Treg与vWF、PⅢNP、ⅠCTP水平及皮肤积分、疾病活动指数相关性。结果 SSc患者外周血Th17比例、Th17/Treg及血清IL-17含量高于健康组,而Treg比例及血清IL-10含量低于健康组,差异均有统计学意义(P<0.05,P<0.01)。两组治疗后Th17比例、Th17/Treg、IL-17含量、vWF及PⅢNP水平均较治疗前降低(P<0.05,P<0.01),Treg比例、IL-10含量升高(P<0.05,P<0.01),治疗组ⅠCTP水平较治疗前升高(P<0.05)。除Th17比例外,治疗组各指标水平改善优于对照组(P<0.05,P<0.01)。SSc患者治疗前后Th17/Treg与vWF、PⅢNP水平及皮肤积分、疾病活动指数呈正相关性(P<0.01),与ⅠCTP水平无明显相关性(P>0.05)。结论温阳化浊通络方能够通过调节SSc患者外周血Th17/Treg细胞失衡、降低vWF和PⅢNP水平、升高ⅠCTP水平而发挥治疗作用。

Objective To observe the effect of Wenyang Huazhuo Tongluo Recipe （WYHZTLR） on the proportion of T helper 17 cells （Th17）/regulatory T cells （Treg）, and serum levels of IL-17 and IL- 10 in peripheral blood of systemic sclerosis （SSc） patients with yang qi insufficiency and turbidity induced collaterals blockage syndrome （YQITICBS）. Methods Totally 82 SSc patients were randomly assigned to the Western medicine group （as the control group） and the integrated Chinese and Western medicine group （as the treatment group）, 41 cases in each group. All patients took methotrexate （MTX） tablet and prednisone tablet. Patients in the treatment group additionally took WYHZTLR. The treatment course for all was six consecutive months. Besides, another 70 healthy volunteers were recruited as a healthy control group （as the healthy group）. Percentages of Th17 and Treg in peripheral blood were detected by flow cytometry. Serum levels of IL-17, IL-10, von Willebrand factor （vWF）, aminoterminal propeptide of type Ⅲ procollagen （ PⅢNP）, and cross-linked carboxyterminal telopeptide of type [ collagen （ Ⅰ CTP） were measured by double antibody sandwich ELISA. The correlations between Th17/Treg and levels of vWF, PⅢNP,Ⅰ CTP, skin score, and disease activity index were observed by Pearson cor relation analysis. Results The percentage of Th17 in peripheral blood, ratios of Th17/Treg, and the serum level of IL-17 were significantly higher, but the percentage of Treg and the serum level of IL-10 were significantly lower in SSc patients, when compared with those of the healthy group （P 〈0.05, P 〈0.01 ）. Compared with the same group before treatment, the percentage of Fh17, ratios of Th17/Treg, and levels of IL-17, vWF, and PⅢNP all decreased in the two groups after treatment （P 〈0.05, P 〈0.01 ）,but the percentage of Treg and the IL-10 level increased （P 〈0.05, P 〈0.01 ）. Meanwhile,the level of I CTP was higher in the treatment group after treatment （P 〈0.05）. The improvement of all indices except the percentage of Th17 was more obvious in the treatment group than in the control group （P 〈0.05, P 〈0.01 ）. The ratio of Th17/Treg was positively correlated with levels of vWF, PⅢNP, skin score, and disease activity index before and after treatment respectively （P 〈0.01 ）, but with no obvious correlation with the level of Ⅰ CTP （P 〉0.05）. Conclusion WYHZTLR could achieve its therapeutic effect on SSc patients by regulating Th17/Treg imbalance, lowering levels of vWF and PⅢNP, and elevating the level of Ⅰ CTP.