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江西中医药大学研究生院,南昌 330004
南昌大学第二附属医院中医科,江西南昌 330006
李逸成,江西中医药大学硕士研究生。E-mail:847612235@qq.com
✉熊俊,教授。E-mail:xiongjun196071@163.com
收稿:2024-05-17,
纸质出版:2025-07-12
移动端阅览
李逸成, 熊俊, 林萌, 等. 基于PI3K/AKT信号通路探讨热敏灸“肺俞”对AR模型大鼠免疫炎性反应的影响[J]. 中国针灸, 2025,45(7):957-966.
LI Yicheng, XIONG Jun, LIN Meng, et al. Effect of heat-sensitive moxibustion at "Feishu" (BL13) on immunoinflammatory response in allergic rhinitis rats based on PI3K/AKT signaling pathway[J]. Chinese Acupuncture & Moxibustion, 2025, 45(7): 957-966.
李逸成, 熊俊, 林萌, 等. 基于PI3K/AKT信号通路探讨热敏灸“肺俞”对AR模型大鼠免疫炎性反应的影响[J]. 中国针灸, 2025,45(7):957-966. DOI: 10.13703/j.0255-2930.20240517-k0004.
LI Yicheng, XIONG Jun, LIN Meng, et al. Effect of heat-sensitive moxibustion at "Feishu" (BL13) on immunoinflammatory response in allergic rhinitis rats based on PI3K/AKT signaling pathway[J]. Chinese Acupuncture & Moxibustion, 2025, 45(7): 957-966. DOI: 10.13703/j.0255-2930.20240517-k0004.
目的:
2
基于磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)信号通路观察热敏灸“肺俞”对变应性鼻炎(AR)大鼠免疫炎性反应的影响,探讨其作用机制。
方法:
2
将32只雄性SD大鼠随机分为空白组(6只)、造模组(26只),造模组采用卵清白蛋白(OVA)全身及局部攻击致敏法制备AR模型。将造模成功大鼠随机分为模型组(6只)、药物组(6只)、艾灸组(14只)。艾灸组大鼠予悬灸双侧“肺俞”干预,每次40 min,每日1次,连续21 d,同时检测并记录体温和尾温。若艾灸组大鼠干预期间体温及尾温均升高>1 ℃则判定为出现腧穴热敏化效应,分入热敏灸组(8只,为保证样本量一致,随机选取6只),若未出现腧穴热敏化效应则分入传统艾灸组(6只)。药物组予丙酸氟替卡松鼻喷雾剂滴鼻,每侧8 μL,每日1次,共21 d。观察大鼠造模后及干预后鼻炎症状行为学评分及造模后血清免疫球蛋白E(IgE)含量。干预后,采用HE染色法观察大鼠鼻黏膜组织形态;免疫组化法检测大鼠鼻黏膜胸腺基质淋巴细胞生成素(TSLP)阳性表达;ELISA法检测大鼠血清白细胞介素(IL)-4、IL-5、IL-13、干扰素-γ(IFN-γ)含量;Western blot法检测大鼠鼻黏膜肿瘤坏死因子受体超家族成员4(OX40)、磷酸化蛋白激酶B(p-AKT)、磷酸化磷脂酰肌醇3-激酶(p-PI3K)蛋白表达。
结果:
2
造模后,造模组鼻炎症状行为学评分及血清IgE含量较空白组升高(
P
<
0.01),提示AR模型建立成功。干预后,与空白组比较,模型组鼻炎症状行为学评分升高(
P
<
0.01);鼻黏膜结构紊乱,炎性细胞浸润严重;鼻黏膜TSLP阳性表达升高(
P
<
0.01);血清IgE、IL-4、IL-5、IL-13含量均升高(
P
<
0.01),IFN-γ含量降低(
P
<
0.01);鼻黏膜OX40、p-AKT、p-PI3K蛋白表达升高(
P
<
0.05)。与模型组比较,药物组、传统艾灸组、热敏灸组大鼠鼻炎症状行为学评分降低(
P
<
0.01);鼻黏膜结构、炎性细胞浸润及血管扩张情况均有不同程度改善;鼻黏膜TSLP阳性表达均降低(
P
<
0.01);血清IgE、IL-4、IL-5、IL-13含量均降低(
P
<
0.05),IFN-γ含量均升高(
P
<
0.05)。与模型组比较,药物组及传统艾灸组鼻黏膜p-AKT蛋白表达降低(
P
<
0.05),热敏灸组鼻黏膜OX40、p-AKT、p-PI3K蛋白表达均降低(
P
<
0.05)。与药物组比较,热敏灸组鼻黏膜TSLP阳性表达降低(
P
<
0.05)。与传统艾灸组比较,热敏灸组和药物组大鼠血清IL-13含量降低、IFN-γ含量升高(
P
<
0.05);药物组鼻黏膜p-PI3K蛋白表达降低(
P
<
0.05);热敏灸组大鼠鼻黏膜TSLP阳性表达及OX40、p-PI3K蛋白表达降低(
P
<
0.05)。
结论:
2
热敏灸“肺俞”可减轻AR模型大鼠临床症状,改善鼻黏膜炎性细胞浸润及毛细血管扩张情况,抑制免疫炎性反应,可能通过调节PI3K/AKT信号通路实现。
Objective
2
To observe the effect of heat-sensitive moxibustion at "Feishu" (BL13) on immunoinflammatory response in rats with allergic rhinitis (AR) based on phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway
so as to explore its underlying mechanism.
Methods
2
Thirty-two male SD rats were randomly divided into a blank group (6 rats) and a modeling group (26 rats). In the modeling group
AR model was prepared using systemic and local attack sensitization method with ovalbumin. The successfully-modeled rats were randomized into a model group (6 rats)
a medication group (6 rats) and a moxibustion group (14 rats). In the moxibustion group
the suspending moxibustion was operated at bilateral "Feishu" (BL13)
40 min each time
once daily
for 21 consecutive days; during which
the temperature of the body and tail was recorded. During intervention
if the temperature of the body and tail increased by >1 ℃
the heat-sensitive reaction at the point was determined in the rats of the moxibustion group
and these rats were collected in a heat-sensitive moxibustion group (8 rats involved and 6 rats of them were randomly collected to ensure the sample-size consistency); and those without heat-sensitive moxibustion reaction were assigned to a traditional moxibustion group (6 rats). In the medication group
fluticasone propionate nasal spray was applied
8 μL on each side
once daily and for 21 days. The behavioral score for AR symptoms after modeling and intervention
and the content of serum immunoglobulin E (IgE) after modeling were observed. After intervention
the histological morphology of the nasal mucosa was observed using HE staining
the positive expression of thymic stromal lymphopoietin (TSLP) in the nasal mucosa was detected using immunohistochemistry
the levels of IgE
interleukin (IL)-4
IL-5
IL-13 and interferon-γ (IFN-γ) were detected by ELISA
and the protein expression of the member 4 of tumor necrosis factor receptor superfamily (OX40)
phosphorylated protein kinase B (p-AKT)
phosphorylated phosphatidylinositol 3-kinase (p-PI3K) in nasal mucosa was detected by Western blotting.
Results
2
After modeling
the behavioral score of AR symptoms and serum IgE level in the modeling group were higher than those of the blank group (
P
<
0.01)
suggesting the success of AR modeling. After intervention
compared with the blank group
the behavioral score of AR symptoms was increased (
P
<
0.01);the nasal mucosa structure was disordered
the inflammatory infiltration was severe; the positive expression of TSLP in the nasal mucosa increased (
P
<
0.01)
the levels of serum IgE
IL-4
IL-5
and IL-13 elevated (
P
<
0.01)
and the level of IFN-γ decreased (
P
<
0.01); and the protein expression of OX40
p-AKT
and p-PI3K in the nasal mucosa increased (
P
<
0.05) in the model group. Compared with the model group
the behavioral score of AR symptoms was reduced (
P
<
0.01); the nasal mucosa structure
inflammatory infiltration
and vascular dilation were ameliorated to varying degrees; the positive expression of TSLP in the nasal mucosa decreased (
P
<
0.01); the content of serum IgE
IL-4
IL-5
and IL-13 decreased (
P
<
0.05)
and that of IFN-γ increased (
P
<
0.05) in the med
ication
traditional moxibustion
and heat-sensitive moxibustion groups. Compared with the model group
the protein expression of p-AKT was reduced in the medication and traditional moxibustion groups (
P
<
0.05)
the protein expression of OX40
p-AKT
and p-PI3K in the nasal mucosa decreased in the heat-sensitive moxibustion group (
P
<
0.05). When compared with the medication group
the positive expression of TSLP in the nasal mucosa was reduced (
P
<
0.05) in the heat-sensitive moxibustion group. In comparison with the traditional moxibustion group
the content of serum IL-13 was reduced and the content of IFN-γ elevated in the heat-sensitive moxibustion and the medication groups (
P
<
0.05)
the protein expression of p-PI3K reduced in the medication group (
P
<
0.05)
and the positive expression of TSLP and the protein expression of OX40 and p-PI3K in the nasal mucosa were reduced in the heat-sensitive moxibustion group (
P
<
0.05).
Conclusion
2
Heat-sensitive moxibustion at "Feishu" (BL13) can alleviate the symptoms of AR rats
ameliorate the inflammatory infiltration and telangiectasia of nasal mucosa
and inhibit immunoinflammatory response
which may be obtained by regulating PI3K/AKT signal pathway.
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