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1. 河南中医药大学中医学院(仲景学院)
2. 河南中医药大学
纸质出版日期:2022
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华金双, 李菡洁, 陈放, 等. 针刺预处理结合磷脂酰肌醇激酶阻断剂对哮喘大鼠气道炎性反应的影响[J]. 针刺研究, 2022,47(9):778-784.
HUA Jin-shuang, LI Han-jie, CHEN Fang, et al. Effect of acupuncture preconditioning combined with PI3K blocker LY294002 on airway inflammation in asthmatic rats[J]. Acupuncture research, 2022, 47(9): 778-784.
华金双, 李菡洁, 陈放, 等. 针刺预处理结合磷脂酰肌醇激酶阻断剂对哮喘大鼠气道炎性反应的影响[J]. 针刺研究, 2022,47(9):778-784. DOI: 10.13702/j.1000-0607.20210650.
HUA Jin-shuang, LI Han-jie, CHEN Fang, et al. Effect of acupuncture preconditioning combined with PI3K blocker LY294002 on airway inflammation in asthmatic rats[J]. Acupuncture research, 2022, 47(9): 778-784. DOI: 10.13702/j.1000-0607.20210650.
目的:观察针刺预处理结合磷脂酰肌醇激酶(PI3K)阻断剂LY294002对哮喘大鼠肺组织中PI3K、蛋白激酶B(Akt)蛋白和mRNA表达及血清中白细胞介素(IL)-12、IL-13含量的影响,探讨针刺预处理结合PI3K阻断剂LY294002对哮喘大鼠的预保护作用。方法:Wistar大鼠按照随机数字表法分为空白组、模型组、针刺预处理+空白组、针刺预处理组、针刺预处理+LY294002组、LY294002组,每组10只。采用致敏、激发的方法制备哮喘模型。针刺预处理+LY294002组、LY294002组每次激发前给予LY294002溶液雾化阻断30 min;针刺预处理组、针刺预处理+LY294002组于造模前选择“肺俞”“大椎”“风门”针刺20 min
每日1次,治疗7 d。采用HE染色法观察各组大鼠肺组织病理形态学变化;ELISA法检测各组大鼠血清IL-12、IL-13含量;免疫组织化学法检测肺组织PI3K、Akt蛋白表达情况;荧光定量PCR法检测肺组织中Akt mRNA的表达。结果:模型组大鼠支气管及肺间质内可见大量炎性因子浸润,支气管管腔缩窄明显,渗出物增多。与模型组相比,针刺预处理组、针刺预处理+LY294002组、LY294002组大鼠炎性浸润程度减轻,支气管腔缩窄程度较轻,黏液分泌减少,肺泡结构轻微不完整。与空白组比较,模型组大鼠血清IL-12含量明显降低(P<0.01)
IL-13含量明显升高(P<0.01);与模型组比较,针刺预处理组、针刺预处理+LY294002组、LY294002组大鼠血清IL-12含量均明显升高(P<0.01
P<0.05)
IL-13含量均明显降低(P<0.01)。与空白组比较,模型组大鼠肺组织PI3K、Akt蛋白表达及Akt mRNA表达明显升高(P<0.01);与模型组比较,针刺预处理组、针刺预处理+LY294002组、LY294002组大鼠肺组织PI3K、Akt蛋白及Akt mRNA表达均明显降低(P<0.05
P<0.01)。与针刺预处理组比较,除Akt mRNA外,针刺预处理+LY294002组上述指标均明显改善(P<0.01)。空白组与针刺预处理+空白组各指标差异均无统计学意义(P>0.05)。结论:针刺预处理可以抑制哮喘大鼠气道炎性反应,其中针刺预处理结合LY294002作用最佳,其机制可能是通过调控PI3K、Akt蛋白表达及炎性因子分泌而发挥对哮喘的预防作用。
Objective To observe the effect of acupuncture preconditioning combined with PI3K blocker LY294002 on the expression of PI3K and Akt proteins and genes in the lung tissue and the contents of serum IL-12 and IL-13 in asthmatic rats
so as to explore its preprotective mechanism underlying improving asthma. Methods Sixty male Wistar rats were randomly divided into blank control
model
acupuncture pretreatment + blank
acupuncture pretreatment
acupuncture pretreatment + LY294002 and LY294002 groups(n=10 in each group). The asthma model was established by intraperitoneal injection of mixture solution of OVA and Al(OH)_3 and followed inhalation of 1%OVA for 30 min
once daily for 7 days. Rats of the blocker groups received inhalation of atomized LY294002 solution for 30 min before inhalation of 1% OVA
and acupuncture was applied to “Feishu”(BL13)
“Dazhui”(GV14) and “Fengmen”(BL12) for 20 min
once daily for 7 days before modeling. H.E. staining was used to assess histopathological changes of the lung tissue
and ELISA was used to detect the contents of serum IL-12 and IL-13. The immunoactivity of PI3K and Akt and expression of Akt mRNA of the lung tissue were detected by using immunohistochemistry and fluorescence quantitative real-time PCR
separately.Results Compared with the blank control group
the content of serum IL-12 was significantly decreased(P<0.01)
and the content of serum IL-13
the expression levels of PI3K
Akt protein and Akt mRNA were remarkably increased(P<0.01) in the model group. In comparison with the model group
the content of serum IL-12 in the pretreatment
pretreatment + LY294002 and LY294002 groups was significantly increased(P<0.01
P<0.05)
while the content of IL-13 and the expression levels of PI3K
Akt protein and Akt mRNA were considerably decreased(P<0.01
P<0.05) in the acupuncture pretreatment
acupuncture pretreatment+LY294002 and LY294002 groups. The therapeutic effect of acupuncture pretreatment+LY294002 was obviously superior to that of simple acupuncture pretreatment and LY294002(except PI3K and Akt in the LY294002 group) in up-regulating serum IL-12 level
and in down-regulating serum IL-13
and PI3K and Akt protein levels in the lung tissue(P<0.01). H.E. staining showed severe inflammatory factor infiltration in the bronchus and pulmonary interstitium
and obvious bronchial lumen narrowing with increased exudate in rats of the model group
which was relatively milder in rats of the acupuncture pretreatment
acupuncture pretreatment+LY294002 and LY294002 groups. There were no significant diffe-rences between blank control and pretreatment+blank groups in all of the above indicators(P>0.05). Conclusion Acupuncture preconditioning can inhibit airway inflammation in asthmatic rats
which may be associated with its functions in down-regulating the levels of pulmonary PI3K and Akt and serum IL-13 and up-regulating the content of serum IL-12. Acupuncture preconditioning combined with LY294002 has the best effect.
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