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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,长沙 410208
Received:08 January 2025,
Accepted:26 March 2025,
Published Online:27 March 2025,
Published:20 July 2025
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王千怀,屈一平,徐小珊等.隔药饼灸对免疫抑制大鼠BAFF及CXCL13信号通路的影响[J].中国实验方剂学杂志,2025,31(14):86-95.
WANG Qianhuai,QU Yiping,XU Xiaoshan,et al.Effect of Herb-partitioned Moxibustion on BAFF and CXCL13 Signaling Pathway in Immunosuppressed Rats[J].Chinese Journal of Experimental Traditional Medical Formulae,2025,31(14):86-95.
王千怀,屈一平,徐小珊等.隔药饼灸对免疫抑制大鼠BAFF及CXCL13信号通路的影响[J].中国实验方剂学杂志,2025,31(14):86-95. DOI: 10.13422/j.cnki.syfjx.20250640.
WANG Qianhuai,QU Yiping,XU Xiaoshan,et al.Effect of Herb-partitioned Moxibustion on BAFF and CXCL13 Signaling Pathway in Immunosuppressed Rats[J].Chinese Journal of Experimental Traditional Medical Formulae,2025,31(14):86-95. DOI: 10.13422/j.cnki.syfjx.20250640.
目的
2
观察隔药饼灸干预对免疫抑制大鼠B细胞激活因子(BAFF)及趋化因子13(CXCL13)、趋化因子5(CXCR5)信号通路的影响,阐释隔药饼灸对机体B细胞作用可能的分子机制。
方法
2
将40只SD大鼠随机分为4组:空白组、模型组、隔药饼灸组及艾条灸组,每组10只。除空白组外,其余各组采用腹腔注射环磷酰胺,以30 mg·kg
-1
剂量对其进行模型制备,持续3 d。隔药饼灸组与艾条灸组选择“中脘”“神阙”“关元”“足三里(双)”进行干预,连续10 d。每日记录大鼠体质量,并在干预结束后次日,对动物进行称体质量、麻醉,取静脉血、动物脾脏,计算脾脏指数,酶联免疫吸附测定法(ELISA)检测血清中
γ
干扰素(IFN-
γ
)、分化簇19(CD19)、分化簇79(CD79)、分化簇27(CD27)、免疫球蛋白G(IgG)、免疫球蛋白M(IgM)、白细胞介素-4(IL-4)及白细胞介素-21(IL-21)含量,并计算Th1/Th2值;采用苏木素-伊红(HE)染色观察脾脏组织形态学变化;实时荧光定量聚合酶链式反应(Real-time PCR)检测脾组织中免疫球蛋白样转录子4(ILT4)、BAFF蛋白表达量;蛋白免疫印迹法(Western blot)检测脾
组织中CXCL13、CXCR5蛋白表达。
结果
2
与空白组比较,模型组大鼠体质量下降明显(
P
<
0.05),脾脏指数显著上升(
P
<
0.01),血清中CD19、CD27、CD79、IL-21含量显著减少(
P
<
0.01),IFN-
γ
含量和Th1/Th2值显著升高,IL-4含量明显降低(
P
<
0.05),IgG和IgM的含量均明显下降(
P
<
0.05),脾脏组织细胞排列紊乱,白髓和红髓之间界限不清晰,脾小结模糊,ILT4 mRNA表达显著升高(
P
<
0.01),BAFF mRNA表达显著降低(
P
<
0.01),CXCL13、CXCR5蛋白表达显著降低(
P
<
0.01);与模型组比较,艾灸组和隔药饼灸组大鼠体质量明显升高(
P
<
0.05),脾脏指数显著下降(
P
<
0.01),血清中CD19、CD27、CD79、IL-21、IL-4含量显著升高(
P
<
0.01),IFN-
γ
含量、Th1/Th2值明显下降(
P
<
0.05),IgG和IgM含量均明显上升(
P
<
0.05),脾脏淋巴细胞弥漫性增生和浸润现象明显缓解,且白髓和红髓界限较清晰,ILT4 mRNA表达显著降低(
P
<
0.01),BAFF mRNA表达显著升高(
P
<
0.01),CXCL13、CXCR5蛋白表达显著升高(
P
<
0.01),隔药饼灸组与艾条灸组比较,除体质量、IL-4、IgG外,其余观测指标两组间均有明显差异(
P
<
0.05,
P
<
0.01)。
结论
2
隔药饼灸可以通过调节机体B细胞、CXCL13、CXCR5及细胞激活因子BAFF改善机体免疫状态,提高机体免疫功能。
Objective
2
To investigate the effects of herb-partitioned moxibustion on B-cell activating factor (BAFF) and the chemokine (C-X-C motif) ligand 13 (CXCL13)/C-X-C motif chemokine receptor 5 (CXCR5) signaling pathway in immunosuppressed rats and elucidate the potential molecular mechanisms underlying the regulatory effects of herb-partitioned moxibustion on B cells in the body.
Methods
2
Forty SD rats were randomly allocated into four groups (
n
=10): blank, model, herb-partitioned moxibustion, and moxa stick moxibustion. The model was established by intraperitoneal injection of cyclophosphamide at 30 mg·kg
-1
for 3 consecutive days in other groups except the blank group. Shenque (CV8), Guanyuan (CV4), Zhongwan (CV12), and Zusanli (ST36, bilateral) were selected for interventi
on in the two moxibustion groups for 10 consecutive days. The body weight of each rat was recorded every day. On the day following the end of intervention, the rats were weighed and anesthetized. Then, venous blood and spleen samples were collected, and the spleen index was calculated. The enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of interferon-
γ
(IFN-
γ
), cluster of differentiation 19 (CD19), cluster of differentiation 79 (CD79), cluster of differentiation 27 (CD27), immunoglobulin G (IgG), immunoglobulin M (IgM), interleukin-4 (IL-4), and interleukin-21 (IL-21) in the serum, and the Th1/Th2 ratio was calculated. Hematoxylin-eosin staining was used to examine the morphological alterations in the spleen tissue. Real-time quantitative polymerase chain reaction(Real-time PCR) was utilized to determine the mRNA levels of immunoglobulin-like transcript 4 (ILT4) and B cell activating factor (BAFF) in the spleen tissue. Western blot was employed to measure the protein levels of CXCL13 and CXCR5 in the spleen tissue.
Results
2
Compared with the blank group, the model group exhibited a decrease in body weight (
P
<
0.05), an increase in spleen index (
P
<
0.01), cell disarrangement in the spleen tissue, an obscure boundary between the white pulp and the red pulp, and fuzzy spleen nodules. In addition, the modeling led to declined serum levels of CD19 (
P
<
0.01), CD27 (
P
<
0.01), CD79 (
P
<
0.01), IL-21 (
P
<
0.01), IL-4 (
P
<
0.05), and IgG and IgM (
P
<
0.05), elevated IFN-
γ
and Th1/Th2 levels (
P
<
0.05), up-regulated mRNA level of ILT4 (
P
<
0.01), and down-regulated mRNA level of BAFF (
P
<
0.01) and protein levels of CXCL13 and CXCR5 (
P
<
0.01). Compared with the model group, the herb-partitioned moxi
bustion and moxa stick moxibustion groups showed an increase in body weight (
P
<
0.05), a decrease in spleen index (
P
<
0.01), alleviated diffuse proliferation and infiltration of lymphocytes in the spleen, and a clear boundary between the white pulp and the red pulp. Furthermore, both moxibustion groups showcased elevated serum levels of CD19 (
P
<
0.01), CD27 (
P
<
0.01), CD79 (
P
<
0.01), IL-21 (
P
<
0.01), IL-4 (
P
<
0.01), and IgG and IgM (
P
<
0.05), declined IFN-
γ
and Th1/Th2 levels (
P
<
0.05), down-regulated mRNA level of ILT4 (
P
<
0.01), and up-regulated mRNA level of BAFF (
P
<
0.01) and protein levels of CXCL13 and CXCR5 (
P
<
0.01). The herb-partitioned moxibustion group and the moxa stick moxibustion group showed differences in other indicators (
P
<
0.05,
P
<
0.01) except the body weight, IL-4, and IgG.
Conclusion
2
Herb-partitioned moxibustion can improve the immune status and immune function by regulating B cells, CXCL13, CXCL5, and BAFF.
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