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谭怡丝,谭劲,朱可可,符静,王宗康.加味丹玄口康对口腔黏膜下纤维化大鼠免疫细胞因子TNF-α、IL-1β、IL-6以及IFN-γ水平的影响[J].湖南中医药大学学报,2022,42(10):1663-1668[点击复制] |
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加味丹玄口康对口腔黏膜下纤维化大鼠免疫细胞因子TNF-α、IL-1β、IL-6以及IFN-γ水平的影响 |
谭怡丝,谭劲,朱可可,符静,王宗康 |
(湖南中医药大学, 湖南 长沙 410208;湖南中医药大学第一附属医院, 湖南 长沙 410007) |
摘要: |
目的 观察加味丹玄口康对口腔黏膜下纤维化(oral submucous fibrosis, OSF)大鼠血清乳酸脱氢酶(lactate dehydrogenase, LDH)、肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)、白细胞介素-1β(interleukin-1β, IL-1β)、白细胞介素-6(interleukin-6, IL-6)及γ干扰素(interferon-γ, IFN-γ)水平的影响,进一步验证加味丹玄口康治疗OSF的疗效机制。方法 60只雄性SD大鼠随机均分为5组(正常组、模型组及加味丹玄口康低、中、高剂量组)。正常组自然饲养,其余4组采用颊黏膜下注入槟榔提取物(areca nut extract, ANE)后再用自制刷蘸ANE刷颊黏膜的方法造模8周。加味丹玄口康高、中、低剂量组通过灌服4、8、12 g/kg剂量的加味丹玄口康进行干预,模型组灌以10 mL/(kg·d)蒸馏水,均连续8周。实验结束后测定大鼠开口度,采用HE染法观察各组大鼠口腔颊黏膜病理形态,采用ELISA法检测各组大鼠血清中LDH、TNF-α、IL-1β、IL-6和IFN-γ水平。结果 与正常组比较,模型组大鼠颊黏膜出现明显的纤维化表现,张口度和血清IFN-γ水平降低(P<0.05),血清LDH、TNF-α、IL-1β、IL-6水平显著升高(P<0.05)。与模型组比较,加味丹玄口康高、中、低剂量组张口度、血清IFN-γ水平均升高(P<0.05),血清LDH、TNF-α、IL-1β、IL-6水平明显降低(P<0.05)。与加味丹玄口康低剂量组比较,加味丹玄口康中剂量组血清IFN-γ水平升高(P<0.05),血清LDH、IL-1β、IL-6降低(P<0.05),加味丹玄口康高剂量组张口度和血清IFN-γ水平升高(P<0.05),血清LDH、TNF-α、IL-1β、IL-6水平明显降低(P<0.05)。与加味丹玄口康中剂量组比较,加味丹玄口康高剂量组张口度和血清IFN-γ水平均明显升高(P<0.05),血清LDH、TNF-α、IL-1β、IL-6水平明显降低(P<0.05)。OSF大鼠血清TNF-α、IL-1β、IL-6水平与张口度均呈高度负相关,与血清LDH水平呈高度正相关(P<0.01),血清IFN-γ水平与张口度呈高度正相关,与血清LDH水平呈负相关(P<0.01)。结论 加味丹玄口康能通过降低促纤维化免疫因子TNF-α、IL-1β、IL-6的分泌,增加抑制纤维化免疫因子IFN-γ的分泌,影响机体免疫功能,并能增加张口度及降低OSF生物标志物LDH水平,进而改善口腔黏膜下纤维化,控制OSF病情进展。 |
关键词: 加味丹玄口康 口腔黏膜下纤维化 乳酸脱氢酶 肿瘤坏死因子-α 白细胞介素-1β 白细胞介素-6 γ干扰素 |
DOI:10.3969/j.issn.1674-070X.2022.10.012 |
投稿时间:2022-04-18 |
基金项目:国家自然科学基金项目(81874496)。 |
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Effects of Modified Danxuan Koukang on immune cytokine levels of TNF-α, IL-1β, IL-6, and IFN-γ in rats with oral submucosal fibrosis |
TAN Yisi,TAN Jin,ZHU Keke,FU Jing,WANG Zongkang |
(Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
Abstract: |
Objective To observe the effects of Modified Danxuan Koukang on serum levels of lactate dehydrogenase (LDH), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-1β), interleukin-6 (IL-6), and interferon-γ (IFN-γ) in rats with oral submucosal fibrosis (OSF), and further verify the therapeutic mechanism of Modified Danxuan Koukang on OSF. Methods A total of 60 male SD rats were randomly divided into 5 groups (normal group, model group, and low-dose, medium-dose, and high-dose groups of Modified Danxuan Koukang). Normal group was naturally fed, and the other 4 groups were treated by injecting acreca nut extract (ANE) into buccal mucosa and then brushing ANE on buccal mucosa with a self-made brush for 8 weeks. The high-dose, medium-dose, and low-dose groups were treated with different doses (4, 8, 12 g/kg) of Modified Danxuan Koukang, and model group was given 10 mL/(kg·d) of distilled water, all for 8 weeks. After the experiment, the mouth opening of rats was tested; HE staining was used to detect the histopathological features in the buccal mucosa of rats; the levels of LDH, TNF-α, IL-1β, IL-6, and IFN-γ in each group were detected by ELISA. Results Compared with normal group, the model group showed obvious fibrosis in the buccal mucosa tissue, significant decrease in mouth opening and IFN-γ (P<0.05), and significant increase in serum levels of LDH, TNF-α, IL-1β, and IL-6 (P<0.05). Compared with model group, the mouth opening and serum level of IFN-γ increased significantly (P<0.05), and the serum levels of LDH, TNF-α, IL-1β, and IL-6 decreased significantly (P<0.05) in high-dose, medium-dose, and low-dose groups of Modified Danxuan Koukang. Compared with low-dose group of Modified Danxuan Koukang, the medium-dose group showed significant increase in serum level of IFN-γ (P<0.05), and significant decrease in serum levels of LDH, IL-1β, and IL-6 (P<0.05). Compared with the low-dose group of Modified Danxuan Koukang, the high-dose group showed significant increase in mouth opening and serum level of IFN-γ (P<0.05), and significant decrease in serum levels of LDH, TNF-α, IL-1β, and IL-6 (P<0.05). Compared with the medium-dose group of Modified Danxuan Koukang, high-dose group showed significant increase in mouth opening and serum level of IFN-γ (P<0.05), and significant decrease in serum levels of LDH, TNF-α, IL-1β, and IL-6 (P<0.05). The serum levels of TNF-α, IL-1β, and IL-6 in OSF rats were highly negatively correlated with mouth opening and highly positively correlated with serum LDH level (P<0.01); serum level of IFN-γ was highly positively correlated with mouth opening but negatively correlated with serum LDH level (P<0.01). Conclusion Modified Danxuan Koukang may reduce the secretion of fibrosis stimulating immune cytokines (TNF-α, IL-1β, and IL-6) and increase the secretion of fibrosis inhibiting immune cytokine (IFN-γ). It could affect the body's immune function, increase mouth opening and reduce biomarker level of LDH in OSF, which further ameliorate symptoms of oral submucosal fibrosis and control the disease progression of OSF. |
Key words: Modified Danxuan Koukang oral submucosal fibrosis lactate dehydrogenase tumor necrosis factor-α interleukin-1β interleukin-6 interferon-γ |
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