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[“足三里”穴埋线及聚乙丙交酯线埋植对健康大鼠皮肤肥大细胞、P物质和组胺的影响]

[Effects of catgut embedding and PGLA embedding at "Zusanli" (ST 36) on skin mast cells, substance P and histamine in healthy rats].

作者信息

Wang Yu-Jia, Li Li-Hong, Hou Xun-Rui, Nie Hong-Fang, Liang Xin, Zhang Qian, Cheng Ling, Ye Ting-Ting

机构信息

Department of Acupuncture and Moxibustion, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China.

Department of Acupuncture and Moxibustion, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China; Clinical Medicine College of Guizhou Medical University.

出版信息

Zhongguo Zhen Jiu. 2023 Aug 12;43(8):944-50. doi: 10.13703/j.0255-2930.20220703-k0002.

Abstract

OBJECTIVE

To observe the effects of catgut embedding and polyglycolic acid/poly-lactic acid (PGLA) embedding at "Zusanli" (ST 36) on the activation of local skin mast cells (MC), and expression of substance P (SP) and histamine (HA), and to explore the mechanism of the temporal stimulation effect of acupoint catgut embedding and provide a foundation for further research on the initiation mechanism of acupoint catgut embedding.

METHODS

One hundred and sixty male SPF-grade SD rats were randomly divided into a blank group (10 rats), a sham-embedding group (50 rats), a catgut group (50 rats), and a PGLA group (50 rats). Each intervention group was further randomly divided into five subgroups according to the time points after intervention: 8 hours, 3 days, 7 days, 14 days, and 21 days, with 10 rats in each subgroup. One-time sham-embedding, catgut embedding and PGLA embedding was given at left "Zusanli" (ST 36) in each intervention group, respectively. The skin and subcutaneous connective tissue of the left "Zusanli" (ST 36) were collected at the corresponding time points after intervention, except for the blank group (only one day before intervention). Toluidine blue staining was used to detect MC count and degranulation, and immunohistochemical staining was used to detect the expression of SP and HA positive cells.

RESULTS

There was no significant difference in MC count between the subgroups of each intervention group and the blank group (>0.05). There was no significant difference in MC count between the subgroups of the catgut group and the PGLA group (>0.05). The MC count in the 8-hour subgroup of PGLA group was higher than that in the 8-hour subgroup of catgut group (<0.05), while the MC count in the 21-day subgroup of PGLA group was lower than that in the 21-day subgroup of catgut group (<0.05). Compared with the blank group, the degranulation rates of MC were increased in the 8-hour and 3-day subgroups of sham-embedding group, 8-hour, 3-day, and 7-day subgroups of catgut group, and 8-hour, 3-day, 7-day, and 14-day subgroups of PGLA group (<0.01, <0.05, <0.001). There was no significant difference in the degranulation rate of MC between the subgroups of the catgut group and the PGLA group (>0.05), and no significant difference in the degranulation rate of MC between the two embedding groups at the same time point (>0.05). Compared with the blank group, the expression of SP positive cells was increased in the 8-hour subgroup of sham-embedding group, 8-hour, 3-day, 7-day, and 14-day subgroups of catgut group, and 3-day, 7-day, and 14-day subgroups of PGLA group (<0.001, <0.05). The expression of SP positive cells in the 7-day subgroup of catgut group was higher than that in the 8-hour subgroup of catgut group (<0.05), while the expression of SP positive cells in the 14-day subgroup of catgut group was lower than that in the 7-day subgroup of catgut group (<0.001). The expression of SP positive cells in the 7-day subgroup of PGLA group was higher than that in the 3-day subgroup of PGLA group (<0.05), while the expression of SP positive cells in the 14-day subgroup of PGLA group was lower than that in the 7-day subgroup of PGLA group (<0.01). There was no significant difference in the expression of SP positive cells between the subgroups of the two embedding groups at the same time point (>0.05). Compared with the blank group, the expression of HA positive cells was increased in the 8-hour, 3-day subgroups of sham-embedding group, 8-hour, 3-day, 7-day, and 14-day subgroups of catgut group, and 8-hour, 3-day, 7-day, 14-day, and 21-day subgroups of PGLA group (<0.001, <0.01, <0.05). The expression of HA positive cells in the 14-day subgroup of catgut group was lower than that in the 7-day subgroup of catgut group (<0.05), while the expression of HA positive cells in the 3-day subgroup of PGLA group was higher than that in the 8-hour subgroup of PGLA group (<0.05), and the expression of HA positive cells in the 14-day subgroup of PGLA group was lower than that in the 7-day subgroup of PGLA group (<0.05). The expression of HA positive cells in the 3-day subgroup of PGLA group was higher than that in the 3-day subgroup of catgut group (<0.05).

CONCLUSION

Catgut and PGLA embedding at "Zusanli" (ST 36) in healthy rats could induce changes in local skin MC, SP, and HA, which may be one of the mechanisms of the temporal stimulation effect after acupoint embedding. There are certain differences between different suture materials. A moderate inflammatory response in the acupoint area, mediated by MC and involving SP and HA, may be one of the initiating factors for the effect of acupoint catgut embedding.

摘要

目的

观察在“足三里”(ST 36)穴位埋线及聚乙醇酸/聚乳酸(PGLA)埋线对局部皮肤肥大细胞(MC)活化、P物质(SP)及组胺(HA)表达的影响,探讨穴位埋线时效刺激效应的机制,为进一步研究穴位埋线的启动机制提供依据。

方法

将160只雄性SPF级SD大鼠随机分为空白组(10只)、假埋线组(50只)、羊肠线组(50只)和PGLA组(50只)。各干预组再根据干预后时间点随机分为5个亚组:8小时、3天、7天、14天和21天,每个亚组10只大鼠。各干预组分别于左侧“足三里”(ST 36)进行一次性假埋线、羊肠线埋线和PGLA埋线。除空白组(仅在干预前一天取材)外,在干预后的相应时间点采集左侧“足三里”(ST 36)的皮肤及皮下结缔组织。采用甲苯胺蓝染色检测MC计数及脱颗粒情况,免疫组织化学染色检测SP和HA阳性细胞的表达。

结果

各干预组亚组与空白组之间MC计数差异无统计学意义(>0.05)。羊肠线组与PGLA组亚组之间MC计数差异无统计学意义(>0.05)。PGLA组8小时亚组的MC计数高于羊肠线组8小时亚组(<0.05),而PGLA组21天亚组的MC计数低于羊肠线组21天亚组(<0.05)。与空白组相比,假埋线组8小时和3天亚组、羊肠线组8小时、3天和7天亚组、PGLA组8小时、3天、7天和14天亚组的MC脱颗粒率升高(<0.01,<0.05,<0.001)。羊肠线组与PGLA组亚组之间MC脱颗粒率差异无统计学意义(>0.05),两个埋线组在同一时间点的MC脱颗粒率差异无统计学意义(>0.05)。与空白组相比,假埋线组8小时亚组、羊肠线组8小时、3天、7天和14天亚组、PGLA组3天、7天和14天亚组的SP阳性细胞表达增加(<0.001,<0.05)。羊肠线组7天亚组的SP阳性细胞表达高于羊肠线组8小时亚组(<0.05),而羊肠线组14天亚组的SP阳性细胞表达低于羊肠线组7天亚组(<0.001)。PGLA组7天亚组的SP阳性细胞表达高于PGLA组3天亚组(<0.05),而PGLA组14天亚组的SP阳性细胞表达低于PGLA组7天亚组(<0.01)。两个埋线组在同一时间点亚组之间的SP阳性细胞表达差异无统计学意义(>0.05)。与空白组相比,假埋线组8小时、3天亚组、羊肠线组8小时、3天、7天和14天亚组、PGLA组8小时、3天、7天、14天和21天亚组的HA阳性细胞表达增加(<0.001,<0.01,<0.05)。羊肠线组14天亚组的HA阳性细胞表达低于羊肠线组7天亚组(<0.05),而PGLA组3天亚组的HA阳性细胞表达高于PGLA组8小时亚组(<0.05),PGLA组14天亚组的HA阳性细胞表达低于PGLA组7天亚组(<0.05)。PGLA组3天亚组的HA阳性细胞表达高于羊肠线组3天亚组(<0.05)。

结论

健康大鼠“足三里”(ST 36)穴位埋线及PGLA埋线可引起局部皮肤MC、SP和HA的变化,这可能是穴位埋线时效刺激效应的机制之一。不同缝合材料之间存在一定差异。由MC介导、涉及SP和HA的穴位局部适度炎症反应可能是穴位埋线效应的启动因素之一。

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