隔药饼灸对免疫抑制大鼠SLAM、PVRL2甲基化及4-1BB调控的影响  

Effects of Herbal Cake Separated Moxibustion on Methylation of SLAM and PVRL2 and Regulation of 4-1BB in Immunosuppressive Rats

作　　者：田岳凤 熊罗节 王慧芳 翟春涛 李玮 TIAN Yuefeng;XIONG Luojie;WANG Huifang;ZHAI Chuntao;LI Wei(The Second Clinical College of Shanxi University of Chinese Medicine,Jinzhong 030619,Shanxi,China;College of Acupuncture,Tuina and Rehabilitation,Hunan University of Chinese Medicine,Changsha 410208,Hunan,China)

机构地区：[1]山西中医药大学第二临床学院,山西晋中030619 [2]湖南中医药大学针灸推拿与康复学院,湖南长沙410208

出　　处：《中华中医药学刊》2025年第1期5-10,I0001-I0005,共11页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82174504,81674062);山西中医药大学基础研究项目(ZJJC2022005)。

摘　　要：目的观察隔药饼灸干预对免疫抑制大鼠信号转导淋巴细胞激活分子(signaling lymphocytic activation molecule,SLAM)、脊髓灰质炎病毒相关受体2基因(poliovirus receptor-related 2,PVRL2)甲基化以及共刺激分子肿瘤坏死因子受体超家族成员9(tumor necrosis factor receptor superfamily member 9,TNFRSF9,又称4-1BB)的影响。方法50只SD大鼠随机分为空白组、免疫抑制模型组、隔药饼灸组、假药饼灸组、中药灌胃组,每组10只。采用腹腔注射环磷酰胺造模,以30 mg/kg剂量对除空白组外其余各组进行模型制备,持续3 d。隔药饼灸组与假药饼灸组均选择“神阙”“关元”“中脘”“足三里(双)”进行干预,连续10 d。中药灌胃组使用六味地黄汤悬浊液灌服,连续10 d。血常规检测各组白细胞(white blood cell,WBC)数量,ELISA法检测血清中CD28、4-1BB、肿瘤坏死因子配体超家族成员9(tumor necrosis factor superfamily member 9,TNFSF9,又称4-1BBL)及白细胞介素-17(interleukin 17,IL-17)含量;免疫荧光法检测脾组织中4-1BB及4-1BBL平均荧光强度;BSP法检测脾组织SLAM及PVRL2。结果血常规结果显示与空白组比较,免疫抑制模型组白细胞显著减少。ELISA结果显示与空白组比较,免疫抑制模型组CD28、4-1BB及IL-17水平均显著升高;与免疫抑制模型组比较,隔药饼灸组CD28、4-1BB及IL-17水平均显著降低。免疫荧光结果显示,与空白组比较,免疫抑制模型组4-1BB及4-1BBL平均荧光强度显著升高;与免疫抑制模型组比较,隔药饼灸组4-1BB及4-1BBL荧光强度均显著降低。甲基化结果显示,与空白组比较,免疫抑制模型组SLAM及PVRL2启动子区甲基化率均降低;与免疫抑制模型组比较,隔药饼灸组SLAM甲基化率升高,但PVRL2甲基化率降低。结论隔药饼灸可能通过调节共刺激分子4-1BB及其配体以及SLAM甲基化水平,改善免疫抑制状态,提高机体免疫功能。Objective To observe the effect of herbal cake separated moxibustion on the methylation of ignaling lymphocytic activation molecule(SLAM)and poliovirus related receptor 2 gene(PVRL2)and the costimulatory molecule[tumor necrosis factor superfamily member 9(4-1BB)]in immunosuppressed rats.Methods Fifty SD rats were randomly divided into blank group,immunosuppressive model group,herbal cake separated moxibustion group,fake cake moxibustion group and Chinese herbal medicine gavage group,with 10 rats in each group.The model was made by intraperitoneal injection of cyclophosphamide,and the other groups except the blank group set up the model with the dose of 30 mg/kg for 3 days.“Shenque(RN8)”“Guanyuan(RN4)”“Zhongwan(RN12)”and“Zusanli(ST36)(double)”were selected for intervention in the herbal cake separated moxibustion group and the fake cake moxibustion group for 10 consecutive days.The Chinese herbal medicine gavage group was given Liuwei Dihuang Decoction(六味地黄汤)suspension for 10 days.The number of white blood cells(WBC)in each group was detected by blood routine,and the contents of CD28,4-1BB,tumor necrosis factor ligand superfamily member 9(4-1BBL)and interleukin-17(IL-17)in serum were detected by ELISA.The average fluorescence intensity of 4-1BB and 4-1BBL in spleen tissue was detected by immunofluorescence.BSP method was used to detect SLAM and PVRL2.Results The blood routine results showed that compared with those of the blank group,the white blood cells in the model group were significantly reduced.ELISA results showed that compared with those of the blank group,the levels of CD28,4-1BB and IL-17 in the immunosuppressive model group were significantly increased.Compared with those of the immunosuppressive model group,the levels of CD28,4-1BB and IL-17 in the herbal cake separated moxibustion group were significantly decreased.The results of immunofluorescence showed that the average fluorescence intensity of 4-1BB and 4-1BBL in the mmunosuppressive model group was significantly higher t

关 键 词：隔药饼灸 免疫抑制 环磷酰胺 肿瘤坏死因子受体超家族成员9 信号转导淋巴细胞激活分子 脊髓灰质炎病毒相关受体2基因 

分 类 号：R245.81[医药卫生—针灸推拿学]