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[火针治疗咪喹莫特诱导的小鼠银屑病样皮损及对信号转导与转录激活因子3(STAT3)通路的影响]

[Effect of fire needling on imiquimod induced psoriasis-like lesion and STAT3 pathway in mice].

作者信息

Feng Fang, Wang Yan, Zhao Jing-Xia, Di Ting-Ting, Meng Yu-Jiao, Chen Zhao-Xia, Qi Cong, Hu Xue-Qing, Wang Ya-Zhuo, Li Ping

机构信息

Clinical Medical College of Beijing University of CM, Beijing 100029, China; Beijing Hospital of TCM Affiliated to Capital Medical University, Beijing Institute of TCM, Beijing Key Laboratory of Clinical Basic Research on Psoriasis, Beijing 100010.

Beijing Hospital of TCM Affiliated to Capital Medical University, Beijing Institute of TCM, Beijing Key Laboratory of Clinical Basic Research on Psoriasis, Beijing 100010.

出版信息

Zhongguo Zhen Jiu. 2022 May 12;42(5):541-8. doi: 10.13703/j.0255-2930.20211112-k0005.

Abstract

OBJECTIVE

To observe the effect of fire needling on psoriasis-like lesion and the signal transducer and activator of transcription 3 (STAT3) pathway in mice and compare the therapeutic effect between different interventions of fire needling therapy (surrounding technique of fire needling, fire needling at "Dazhui" [GV 14] and "Zusanli" [ST 36]).

METHODS

Thirty male BALB/c mice were randomized into a blank group, a model group, a dexamthasone group, a surrounding technique group and an acupoint group, 6 mice in each one. Except the blank group, the mice in the rest groups were established as psoriasis-like lesion model by topical application with imiquimod cream, once daily, consecutively for 8 days. From day 4 to day 8, in the dexamthasone group, gastric infusion with 0.2 mL dexamthasone was administered, once daily. On day 4, 6 and 8, in the surrounding technique group, fire needling was exerted around the skin lesion; and fire needling was applied to "Dazhui" (GV 14) and "Zusanli" (ST 36) in the acupoint group, once a day. The changes in skin lesion on the dorsal parts of mice were observed in each group to score the psoriasis area and severity index (PASI). Using HE staining, the dermal morphological changes and epidermal thickness were observed in the mice of each group. The positive expression of proliferating cell-associated antigen Ki-67 was determined by immunofluorescence. Immunohistochemistry method was used to determine the expressions of , and T cells of skin tissue in each group. Using real-time PCR, the expressions of interleukin (IL)-17, IL-22, tumor necrosis factor α(TNF-α) mRNA were determined. Western blot method was adopted to determine the protein expressions of STAT3 and p-STAT3 in skin tissue in each group.

RESULTS

Compared with the blank group, the scores of each item and the total scores of PASI, as well as the epidermal thickness were all increased in the mice of the model group (<0.01). Except for the erythema scores of the dexamethasone group and the surrounding technique group, the scores of each item and the total scores of PASI, as well as the epidermal thickness were all decreased in each intervention group as compared with the model group (<0.01). The infiltration scores and the total scores in the dexamethasone group and the acupoint group were lower than those in the surrounding technique group respectively (<0.01, <0.05). In comparison with the blank group, Ki-67 positive cell numbers and the numbers of , and T cells in skin tissue were increased in the mice of the model group (<0.01). Ki-67 positive cell numbers and the numbers of , and T cells were reduced in each intervention group as compared with the model group (<0.01), and the numbers of and T cells in the acupoint group were less than the surrounding technique group (<0.01). Compared with the blank group, the mRNA expressions of IL-17, IL-22 and TNF-α and the ratio of p-STAT3 to STAT3 were all increased in the model group (<0.01). The mRNA expressions of IL-17, IL-22 and TNF-α and the ratio of p-STAT3 to STAT3 were all decreased in each intervention group as compared with the model group (<0.01, <0.05). The mRNA expressions of IL-17, IL-22 and TNF-α in the acupoint group, as well as mRNA expression of IL-17 in the surrounding technique group were all lower than the dexamethasone group (<0.01), while, the mRNA expression of IL-22 in the acupoint group was lower than the surrounding technique group (<0.01).

CONCLUSION

Fire needling therapy improves skin lesion severity in imiquimod induced psoriasis-like lesion of the mice, which is probably related to the inhibition of STAT3 pathway activation and the decrease of Th17 inflammatory factors expression. The systemic regulation of fire needling at "Dazhui" (GV 14) and "Zusanli" (ST 36) is superior to the local treatment.

摘要

目的

观察火针治疗对小鼠银屑病样皮损及信号转导与转录激活因子3(STAT3)通路的影响,并比较火针疗法不同干预方式(火针围刺术、针刺“大椎”[GV 14]和“足三里”[ST 36])之间的治疗效果。

方法

将30只雄性BALB/c小鼠随机分为空白组、模型组、地塞米松组、围刺术组和穴位组,每组6只。除空白组外,其余各组小鼠均通过局部涂抹咪喹莫特乳膏建立银屑病样皮损模型,每日1次,连续8天。从第4天至第8天,地塞米松组小鼠每日灌胃0.2 mL地塞米松。在第4、6和8天,围刺术组在皮损周围进行火针治疗;穴位组针刺“大椎”(GV 14)和“足三里”(ST 36),每日1次。观察各组小鼠背部皮肤皮损变化,对银屑病面积和严重程度指数(PASI)进行评分。采用HE染色观察各组小鼠真皮形态变化和表皮厚度。通过免疫荧光法检测增殖细胞相关抗原Ki-67的阳性表达。采用免疫组织化学方法检测各组皮肤组织中 、 和T细胞的表达。采用实时荧光定量PCR检测白细胞介素(IL)-17、IL-22、肿瘤坏死因子α(TNF-α)mRNA的表达。采用蛋白质印迹法检测各组皮肤组织中STAT3和p-STAT3的蛋白表达。

结果

与空白组相比,模型组小鼠各项评分及PASI总分、表皮厚度均增加(<0.01)。与模型组相比,各干预组除地塞米松组和围刺术组的红斑评分外,各项评分及PASI总分、表皮厚度均降低(<0.01)。地塞米松组和穴位组的浸润评分及总分分别低于围刺术组(<0.01,<0.05)。与空白组相比,模型组小鼠皮肤组织中Ki-67阳性细胞数及 、 和T细胞数量增加(<0.01)。与模型组相比,各干预组Ki-67阳性细胞数及 、 和T细胞数量减少(<0.01),穴位组 和T细胞数量少于围刺术组(<0.01)。与空白组相比,模型组IL-17、IL-22和TNF-α的mRNA表达及p-STAT3与STAT3的比值均增加(<0.01)。与模型组相比,各干预组IL-17、IL-22和TNF-α的mRNA表达及p-STAT3与STAT3的比值均降低(<0.01,<0.05)。穴位组IL-17、IL-22和TNF-α的mRNA表达以及围刺术组IL-17的mRNA表达均低于地塞米松组(<0.01),而穴位组IL-22的mRNA表达低于围刺术组(<0.01)。

结论

火针疗法可改善咪喹莫特诱导的小鼠银屑病样皮损严重程度,其机制可能与抑制STAT3通路激活及降低Th17炎性因子表达有关。针刺“大椎”(GV 14)和“足三里”(ST 36)的整体调节作用优于局部治疗。

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