摘要
研究祖师麻及其炮制品对SD大鼠Ⅱ型胶原诱导性关节炎(collagen induced arthritis,CIA)中辅助性T细胞17(Th17)/调节性T细胞(Treg)分化的影响。按体质量将64只SD大鼠随机分为正常组,模型组,祖师麻炙品低、高剂量组,生品低、高剂量组,祖师麻甲素组,雷公藤多苷组。除正常组外,CIA模型在第1次免疫后第7天进行二免,并在二免7 d后灌胃28 d。取材后,采用苏木素-伊红(HE)染色观察CIA大鼠关节滑膜炎症的情况;采用酶联免疫吸附测定法(ELISA)检测血清中转化生长因子-β(TGF-β)、γ干扰素(IFN-γ)、白细胞介素(IL)-2、IL-4、IL-10水平变化;实时荧光定量逆转录PCR(qRT-PCR)和蛋白免疫印迹法检测关节滑膜中分化簇80(B7-1)、分化簇86(B7-2)、分化簇28(CD28)、细胞毒性T淋巴细胞相关抗原-4(CTLA-4)mRNA和蛋白表达;流式细胞术用于检测大鼠关节滑膜中Th17和Treg细胞的比例。结果显示,与正常组相比,模型组大鼠的关节滑膜炎症明显加剧,关节炎指数评分明显上升,免疫器官指数上升(P<0.01);与模型组相比,各给药组均能不同程度地改善大鼠的关节炎症,降低关节评分指数,抑制滑膜增生,降低免疫器官指数;相较于模型组,各给药组中大鼠血清中IL-2、IFN-γ显著下降(P<0.01),TGF-β、IL-4、IL-10显著上升(P<0.01),滑膜中B7-1、CTLA-4 mRNA及蛋白表达显著升高(P<0.01),关节组织中的Th17细胞和Treg细胞的比例出现显著降低(P<0.01)。综上得出祖师麻通过调控B7/CD28/CTLA-4通路以及调节Th17/Treg平衡,抑制CIA大鼠中Th17细胞的表达,促进Treg细胞的表达,进而治疗RA。
This study investigates the effects of Daphnes Cortex and its processed products on the differentiation of Th17/Treg cells in SD rats with typeⅡcollagen-induced arthritis(CIA).Sixty-four SD rats were randomly divided into the normal group(normal),model group(model),fried Daphne giraldii Nitsche low-dose and high-dose groups(FDGN-L group,FDGN-H group),raw D.giraldii Nitsche low-dose and high-dose groups(RDGN-L group,RDGN-H group),daphnetin group(DAPH group),and tripterygium glycosides group(GTW group).Except for the normal group,the CIA model was immunized on the seventh day after the first immunization,and was gavaged for 28 days after the second immunization.After sampling,the inflammation of articular synovial membrane in CIA rats was observed by hematoxylin-eosin(HE)staining;the levels of transforming growth factor-β(TGF-β),interferon-γ(IFN-γ),interleukin(IL)-2,IL-4,and IL-10 in serum were detected by enzyme-linked immunosorbent assay(ELISA);real-time reverse transcription-PCR(qRT-PCR)and Western blot were used to detect the mRNA and protein expressions of cluster of differentiation(CD)80(B7-1),CD 86(B7-2),CD28,and cytotoxic T lymphocyte-associated antigen 4(CTLA-4)in the synovial membrane of rats;flow cytometry was used to detect the proportion of Th17 and Treg cells in the synovial membrane of rats.The results showed that compared with the normal group,the joint synovial inflammation of rats in the model group was significantly aggravated,the arthritis index was significantly increased,and the immune organ index was increased(P<0.01).Compared with the model group,each drug administration group could improve the joint inflammation of rats to varying degrees,reduce the arthritis index,inhibit synovial hyperplasia,and reduce the immune organ index;compared with the model group,the serum levels of IL-2 and IFN-γin each drug administration group were significantly decreased(P<0.01),TGF-β,IL-4,and IL-10 were significantly increased(P<0.01),the mRNA and protein expressions of B7-1 and CTLA-4 in the synovial
作者
赵彩蓉
孟祥龙
王娟
赵泓瑜
韩香
李子昂
张朔生
ZHAO Cai-rong;MENG Xiang-long;WANG Juan;ZHAO Hong-yu;HAN Xiang;LI Zi-ang;ZHANG Shuo-sheng(College of Traditional Chinese Medicine and Food Engineering,Shanxi University of Chinese Medicine,Jinzhong 030619,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第15期4091-4099,共9页
China Journal of Chinese Materia Medica
基金
山西省重点实验室项目(202104010910029)
山西中医药大学2022年度科技创新团队项目(2022TD1014)
山西中医药大学学科建设项目(XN2024093)。
