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河南中医药大学,郑州 450046
谢文英,教授,硕士生导师,从事中医药治疗肺系疾病的临床研究,E-mail:xiewenying1963@163.com
包永生,从事中医药治疗肺系疾病的临床研究,E-mail:85651402@qq.com;
尚立芝,教授,硕士生导师,从事中医药作用机制研究,Tel:0371-86253082,E-mail:lzshang2014@163.com
收稿日期:2019-03-05,
网络出版日期:2019-08-16,
纸质出版日期:2019-12-05
移动端阅览
谢文英, 王俊月, 包永生, 等. 二陈汤加味对慢性阻塞性肺疾病大鼠
Wen-ying XIE, Jun-yue WANG, Yong-sheng BAO, et al. Effect of Modified Erchentang on β2AR/β-arrestin2 Signaling Pathway in Rats with Chronic Obstructive Pulmonary Disease[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(23): 34-40.
谢文英, 王俊月, 包永生, 等. 二陈汤加味对慢性阻塞性肺疾病大鼠
Wen-ying XIE, Jun-yue WANG, Yong-sheng BAO, et al. Effect of Modified Erchentang on β2AR/β-arrestin2 Signaling Pathway in Rats with Chronic Obstructive Pulmonary Disease[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(23): 34-40. DOI: 10.13422/j.cnki.syfjx.20192337.
目的:
2
探讨二陈汤加味对慢性阻塞性肺疾病(COPD)大鼠模型
β
2
肾上腺素受体(
β
2
AR)/
β
-抑制蛋白2(
β
-arrestin2)信号通路的影响,以及血清、肺组织匀浆液和支气管肺泡灌洗液中IL-17表达的影响。
方法:
2
将70只SD大鼠随机分为7组,即正常组、模型组、二陈汤加味高、中、低剂量(40,20,10 g·kg
-1
·d
-1
)组、消咳喘组(5 g·kg
-1
·d
-1
)、二陈汤组(5 g·kg
-1
·d
-1
),每组10只。以烟熏与脂多糖(LPS)气管滴注并用的方法制备大鼠COPD大鼠模型。造模成功后,治疗组分别灌胃给药,正常及模型组则灌等量的生理盐水。采用酶联免疫吸附测定(ELISA)检测大鼠血清、肺组织匀浆液和支气管肺泡灌洗液中白细胞介素-17(IL-17)的含量;采用实时荧光定量PCR(Real-time PCR)检测
β
2
AR的mRNA表达;采用蛋白免疫印迹法(Western blot)检测肺组织
β
2
AR的蛋白表达;采用免疫组化检测肺组织
β
2
AR,
β
-arrestin2的蛋白表达。
结果:
2
与正常组比较,模型组肺组织中
β
2
AR的蛋白表达显著降低(
P
<
0.01);与模型组比较,肺组织中
β
2
AR的蛋白表达显著增加(
P
<
0.01),其中二陈汤加味中剂量组与其他各组均有差异(
P
<
0.05)。与正常组比较,模型组
β
2
AR mRNA表达显著降低(
P
<
0.01);与模型组比较,二陈汤加味高、中、低剂量组、消咳喘组、二陈汤组中
β
2
AR的mRNA表达均显著升高(
P
<
0.01)。其中二陈汤加味中剂量组与其他各给药组比较升高显著(
P
<
0.01)。与正常组比较,模型组血清中IL-17含量显著升高(
P
<
0.01),与模型组比较,各给药组血清中IL-17的含量均受到一定程度的抑制,其中二陈汤加味中剂量组受到的抑制程度尤为明显(
P
<
0.05)。
结论:
2
二陈汤加味可能通过升高
β
2
AR,
β
-arrestin2的表达,并以此降低IL-17的含量,来对抗COPD大鼠的炎症反应,并改善肺功能。
Objective:
2
To explore the effect of modified Erchentang on the signal pathway of
β
2
adrenergicreceptor(
β
2
AR)/arrestin beta 2(
β
-arrestin2) in rats with chronic obstructive pulmonary disease (COPD)
and the expression of interleukin-17(IL-17) in serum
lung homogenate and bronchoalveolar lavage fluid.
Method:
2
Seventy SD rats were randomly divided into seven groups: normal group
model group
modified Erchentang with high
medium and low doses (40
20
10 g·kg
-1
·d
-1
)
Xiaokechuan group (5 g·kg
-1
·d
-1
)
modified Erchentang group (5 g·kg
-1
·d
-1
)
10 rats in each group. The rat model of COPD was established by smoking and lipopolysaccharide (LPS) intratracheal drip. After successful modeling
the treatment group was given intragastric administration
while the normal group and the model group were given the same amount of saline. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of IL-17 in serum
lung homogenate and bronchoalveolar lavage fluid of rats. Real-time fluorescence quantitative PCR (Real-time PCR) was used to detect the expression of
β
2
AR gene. Western blot was used to detect the expression of
β
2
AR protein in lung tissue. The expression of
β
2
AR and
β
-arrestin2 in lung tissue was detected by immunohistochemistry.
Result:
2
Compared with the normal group
the expression of
β
2
AR protein in lung tissue of model group was significantly decreased(
P
<
0.01). Compared with model group
the expression of
β
2
AR protein in lung tissue was significantly increased(
P
<
0.01)
and the middle dose group of modified Erchentang was different from other groups (
P
<
0.05). Compared with normal group
the expression of
β
2
AR in model group was significantly lower(
P
<
0.01)
compared with model group
the expression of
β
2
AR in high
medium and low dose group
Xiaokechuan group and modified Erchentang group was significantly higher(
P
<
0.01). The middle dosage group of modified Erchentang was significantly higher than other groups(
P
<
0.01). Compared with normal group
the serum level of IL-17 in the model group was significantly higher(
P
<
0.01). Compared with model group
the serum level of IL-17 in each group was inhibited to a certain extent
especially in the middle dose group of modified Erchentang (
P
<
0.05).
Conclusion:
2
Modified Erchentang may increase the expression of
β
2
AR and
β
-arrestin2 and decrease the content of IL-17 in order to resist inflammation and improve pulmonary function in COPD rats.
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