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1.贵州中医药大学,贵阳 550025
2.遵义市中医院,贵州 遵义 563000
3.南京中医药大学,南京 210000
Received:27 March 2020,
Published Online:30 July 2020,
Published:05 November 2020
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王庆学,欧阳宇晨,卞尧尧等.千金苇茎汤对香烟烟雾暴露模型大鼠肺组织Th17/Treg分化及其相关细胞因子表达的影响[J].中国实验方剂学杂志,2020,26(21):28-33.
WANG Qing-xue,OUYANG Yu-chen,BIAN Yao-yao,et al.Observation of Effect of Qianjin Weijingtang on Differentiation of Th17/Treg and Expressions of Related Cytokines in Lung Tissues of Model Rats Exposed to Cigarette Smoke[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(21):28-33.
王庆学,欧阳宇晨,卞尧尧等.千金苇茎汤对香烟烟雾暴露模型大鼠肺组织Th17/Treg分化及其相关细胞因子表达的影响[J].中国实验方剂学杂志,2020,26(21):28-33. DOI: 10.13422/j.cnki.syfjx.20201873.
WANG Qing-xue,OUYANG Yu-chen,BIAN Yao-yao,et al.Observation of Effect of Qianjin Weijingtang on Differentiation of Th17/Treg and Expressions of Related Cytokines in Lung Tissues of Model Rats Exposed to Cigarette Smoke[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(21):28-33. DOI: 10.13422/j.cnki.syfjx.20201873.
目的
2
观察千金苇茎汤对香烟烟雾暴露模型大鼠肺组织辅助性T细胞17(Th17)/调节性T细胞(Treg)分化及其相关细胞因子表达的影响。
方法
2
雄性大鼠60只,随机分为正常组,烟雾模型组,乙酰半胱氨酸组、千金苇茎汤高、中、低剂量组,共6组,每组10只。香烟烟雾造模30 d后,药物分别干预30 d,后处死各组大鼠并取组织;实时荧光定量聚合酶链式反应(Real-time PCR)检测维甲酸相关孤儿受体(ROR-
γ
t),叉头状/翼状螺旋转录因子3(Foxp3) mRNA表达,酶联免疫吸附测定(ELISA)检测肺组织白细胞介素-17(IL-17),IL-6,IL-10,转化生长因子-
β
1
(TGF-
β
1
)含量,苏木素-伊红(HE)染色切片观察肺组织病理,流式细胞术检测Treg(Foxp3
+
CD25
+
)比例,Th17(CD4
+
IL-17
+
)比例,Treg/Th17。
结果
2
与正常组比较,模型组ROR-
γ
t mRNA表达显著升高(
P
<
0.01);与模型组比较,千金苇茎汤高、中、低剂量组干预后都能够降低其表达(
P
<
0.05,
P
<
0.01);与正常组比较,模型组Foxp3 mRNA表达明显降低(
P
<
0.05),与模型组比较,千金苇茎汤中低剂量组干预后表达明显升高(
P
<
0.05)。与正常组比较,模型组IL-17,IL-6含量明显升高,IL-10,TGF-
β
1
含量明显降低(
P
<
0.05,
P
<
0.01);与模型组比较,千金苇茎汤高、中、低剂量组干预后能够降低肺组织IL-17,IL-6含量,同时升高IL-10的含量(
P
<
0.05),与Real-time PCR检测的结果趋势一致。流式细胞术检测显示,千金苇茎汤中、低剂量组干预后Th17(CD4
+
IL-17
+
)比例降低,Treg/Th17增加(
P
<
0.05,
P
<
0.01)。
结论
2
千金苇茎汤能调节Treg/Th17,使Th17/Treg达到平衡状态。
Objective
2
The present study was to observe the effect of Qianjin Weijingtang on the differentiation of helper T cells 17 (Th17)/T regulatory cell (Treg) and the expressions of related cytokines in the lung tissues of the model rats exposed to cigarette smoke.
Method
2
Totolly 60 male rats were randomly assigned into six groups (control group, model group, acetylcysteine group and Qianjin Weijingtang high, moddle and low dose groups), with 10 rats in each group. After 30 day's modeling and 30 day's intervention, rats were killed peacefully with their tissues collected. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) was used to detect the expression of retinoic acid associated orphan receptor (ROR-
γ
t) and forkhead/pterygoid helix transcription factor 3 (Foxp3) mRNA, enzyme-linked immunosorbent assay (ELISA) was used to check the concentration of interleukin-17(IL-17), IL-6, IL-10 and transforming growth factor-
β
1
(TGF-
β
1
). Htoxylin eosin (HE) staining was used to observe the pathological changes of lung tissues, while flow cytometry was used to detect Treg(Foxp3
+
CD25
+
), Th17(CD4
+
IL-17
+
) and Treg/Th17 ratio.
Result
2
As compared with the control group, the ROR-
γ
t mRNA expression in model group was higher(
P
<
0.01). As compared with the model group, various doses of Qianjin Weijingtang down-regulated the expression(
P
<
0.05,
P
<
0.01). As compared with the control group, the expression of the Foxp3 mRNA was down-regulated in model group(
P
<
0.05), but was up-regulated in Qianjin Weijingtang middle and low dose groups(
P
<
0.05). Compared with the control group, the concentration of IL-17 and IL-6 in the model group were significantly increased, while the concentration of IL-10 and TGF-
β
1
in the model group were significantly decreased(
P
<
0.05,
P
<
0.01). As compared with the model group, intervention with various doses of Qianjin Weijingtang could help to decrease the concentration of IL-17 and IL-6 in lung tissues, and increase the concentration of IL-10(
P
<
0.05), which were consistent with those of Real-time PCR results. Flow cytometry examination showed that the Th17(CD4
+
IL-17
+
) proportion decreased and Treg/Th17 ratio increased after Qianjin Weijing Tang middle and low dose intervention(
P
<
0.05,
P
<
0.01).
Conclusion
2
Qianjin Weijingtang could regulate Treg/Th17 ratio and help to achieve Th17/Treg balance.
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