Effect of acupuncture combined with bloodletting therapy on mast cell degranulation in urticaria rats

DU Yuzhu; LIANG Yulei; SHI Yu; XUE Yuqiang; LIU Xiang; TIAN Zan; SUN Mingxin; WANG Yanjun

doi:10.13703/j.0255-2930.20240320-k0002

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Effect of acupuncture combined with bloodletting therapy on mast cell degranulation in urticaria rats

Study on Mechanism | 更新时间：2024-11-14

- Effect of acupuncture combined with bloodletting therapy on mast cell degranulation in urticaria rats
- Chinese Acupuncture & Moxibustion Vol. 44, Issue 11, Pages: 1273-1280(2024)
- 作者机构：
  
  1.河北中医药大学针灸推拿学院，石家庄　050200
  3.河北中医药大学基础医学院，石家庄　050200
  2.河北大学附属医院康复科
  4.河北中医药大学第一附属医院皮肤科，石家庄　050011
  5.河北中医药大学第一附属医院针灸科，石家庄　050011
- 作者简介：
- 基金信息：
- DOI：10.13703/j.0255-2930.20240320-k0002
  CLC：
- Received：20 March 2024，
  
  Online First：02 September 2024，
  
  Published：12 November 2024
- 稿件说明：
移动端阅览
DU Yuzhu, LIANG Yulei, SHI Yu, et al. Effect of acupuncture combined with bloodletting therapy on mast cell degranulation in urticaria rats[J]. Chinese Acupuncture & Moxibustion, 2024, 44(11): 1273-1280.
DOI：

DU Yuzhu, LIANG Yulei, SHI Yu, et al. Effect of acupuncture combined with bloodletting therapy on mast cell degranulation in urticaria rats[J]. Chinese Acupuncture & Moxibustion, 2024, 44(11): 1273-1280. DOI： 10.13703/j.0255-2930.20240320-k0002.

摘要

目的：

观察针刺联合刺血疗法对荨麻疹模型大鼠血清炎性因子表达、致敏部位皮肤组织形态及肥大细胞脱颗粒情况的影响，探索其治疗荨麻疹的可能机制。

方法：

SPF级SD大鼠42只，随机选取6只大鼠制备致敏抗血清，剩余36只大鼠随机分为空白组、模型组、阳性药物组、针刺组、刺血组和针刺+刺血组，每组6只，采用被动皮肤过敏反应制备荨麻疹大鼠模型。阳性药物组予1 mL氯雷他定溶液（1 mg∙kg

-1

）灌胃，每日1次；针刺组先予1 mL 0.9%氯化钠溶液灌胃，1 h后进行针刺干预，穴取“百会”“中脘”及双侧“曲池”“血海”，每日1次，每次15 min；刺血组先予1 mL 0.9%氯化钠溶液灌胃，1 h后进行刺血疗法，于“大椎”及双侧“膈俞”点刺放血，每日1次，每穴放血量约0.1 mL；针刺+刺血组先予1 mL 0.9%氯化钠溶液灌胃，1 h后联合采用针刺和刺血干预，每日1次。各组均于第6天开始干预，均干预2周。末次干预结束后，各组大鼠进行抗原激发。ELISA法检测大鼠血清免疫球蛋白E（IgE）、类胰蛋白酶（TPS）、白细胞介素-4（IL-4）、白细胞介素-5（IL-5）、肿瘤坏死因子-α（TNF-α）含量，直尺测量大鼠背部致敏部位皮肤蓝斑直径，HE染色观察大鼠背部致敏部位皮肤组织形态，甲苯胺蓝染色观察大鼠背部致敏部位皮肤肥大细胞脱颗粒情况。

结果：

与空白组比较，模型组大鼠血清IgE、TPS、IL-4、IL-5、TNF-α含量升高（

0.01），背部致敏部位皮肤蓝斑直径增大（

0.01），背部致敏部位皮肤组织肥大细胞脱颗粒率升高（

0.01），模型组大鼠背部致敏部位皮肤组织出现明显炎性细胞浸润及真皮层水肿。与模型组比较，阳性药物组、针刺组、刺血组和针刺+刺血组大鼠血清IgE、TPS、IL-4、IL-5、TNF-α含量降低（

0.01，

0.05），阳性药物组和针刺+刺血组大鼠背部致敏部位皮肤蓝斑直径减小（

0.05），阳性药物组、针刺组、刺血组和针刺+刺血组大鼠背部致敏部位皮肤组织肥大细胞脱颗粒率降低（

0.01），阳性药物组、针刺组、刺血组和针刺+刺血组大鼠背部致敏部位真皮层水肿、炎性细胞浸润情况均有所减轻。阳性药物组和针刺+刺血组大鼠血清IgE、TPS、IL-4、IL-5、TNF-α含量及背部致敏部位皮肤组织肥大细胞脱颗粒率低于针刺组和刺血组（

0.05）。

结论：

针刺联合刺血可抑制荨麻疹模型大鼠背部致敏部位皮肤组织肥大细胞脱颗粒，改善其皮肤组织病理形态，其机制可能与抑制辅助性T细胞2（Th2）分化增殖，调节体液免疫应答有关。

Abstract

Objective

To observe the effects of acupuncture combined with bloodletting on the expression of inflammatory factors in serum

the morphology of sensitized skin tissue and the mast cell degranulation in urticaria rats

and to explore the potential mechanism of this therapy for urticaria.

Methods

Among 42 SD rats of SPF grade

6 rats were randomly collected for the preparation of sensitized antiserum; and the rest 36 rats were randomized into a blank group

a model group

a positive drug group

an acupuncture group

a bloodletting group and a combined treatment group (acupuncture + bloodletting)

6 rats in each one. The rat model of urticaria was established by passive cutaneous anaphylaxis. In the positive drug group

loratadine (1 mg·kg

-1

) by gavage was administered once a day. In the acupuncture group

1 h after gavage with 0.9% NaCl (1 mL)

acupuncture was delivered at "Baihui" (GV 20)

"Zhongwan" (CV 12)

and bilateral "Quchi" (LI 11) and "Xuehai" (SP 10) for 15 min

once daily . In the bloodletting group

1 h after gavage with 0.9% NaCl (1 mL)

bloodletting was operated at "Dazhui" (GV 14) and bilateral "Geshu" (BL 17)

around 0.1 mL of bleeding volume at each point

once daily. In the combined treatment group

1 h after gavage with 0.9% NaCl (1 mL)

the interventions as the acupuncture group and the bloodletting group were adopted

once daily. All the interventions started on day 6 of modeling

lasting 2 weeks. After intervention completion

antigenic stimulation was performed in the rats of each group. Using ELISA

the levels of serum immunoglobulin E (IgE)

tryptase (TPS)

interleukin-4 (IL-4)

interleukin-5 (IL-5)

tumor necrosis factor-α (TNF-α) were detected. The diameter of the blue spots of the sensitized skin on the back was measured with ruler in each rat. The morphology of sensitized skin tissu

e was observed using HE staining

and the degranulation of mast cells was observed using Toluidine blue staining.

Results

Compared with the blank group

in the model group

the levels of serum IgE

TPS

IL-4

IL-5 and TNF-α increased (

0.01)

the diameter of blue spot on the sensitized part of the rat back was larger (

0.01)

the degranulation rate of mast cells was elevated (

0.01)

and there were obvious inflammatory cell infiltration and edema in the dermis of sensitized skin tissue on the rat back. Compared with the model group

the serum levels of IgE

TPS

IL-4

IL-5 and TNF-α were reduced in the positive drug group

the acupuncture group

the bloodletting group and the combined treatment group (

0.01

0.05); skin blue spot diameter was smaller in the positive drug group and the combined treatment group (

0.05); the degranulation rate of mast cells decreased in the positive drug group

the acupuncture group

the bloodletting group and the combined treatment group (

0.01); and the dermal edema

inflammatory infiltration were attenuated in the positive drug group

the acupuncture group

the bloodletting group and the combined treatment group. Compared with the acupuncture group and the bloodletting group

the serum levels of IgE

TPS

IL-4

IL-5 and TNF-α

as well as the degranulation rate of mast cells in the sensitized tissue were lower in the positive drug group and the combined treatment group (

0.05).

Conclusion

Acupuncture combined with bloodletting effectively suppress mast cell degranulation in the sensitized skin tissue on the back of urticaria rats

and ameliorate the histopathological morphology. Its effect mechanism may be related to inhibiting the differentiation and proliferation of helper T cells 2 and regulating the humoral immune response.

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