基于肠道菌群调节研究桂枝加葛根汤治疗颈型颈椎病家兔作用机制
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R285.5;R681.5+3

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南京中医药大学针灸推拿·养生康复学院课题(ZTYK202101)


Study on Mechanism of Guizhi and Gegen Decoction for Neck-Type Cervical Spondy⁃ lopathy in the Rabbits Based on Microbial Flora Regulation
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    摘要:

    目的:基于肠道菌群(IF) 调节研究桂枝加葛根汤治疗颈型颈椎病(NTCS) 家兔的作用机制。方 法:将9 只家兔随机分成空白组、模型组、桂枝加葛根汤组3 组,每组各3 只。空白组不造模,其余2 组均制 成脾虚证NTCS 家兔模型。空白组和模型组不予给药,桂枝加葛根汤组以桂枝加葛根汤药液按10 mL/kg 灌胃。 每天1 次,连续给药7 d。以16 SrRNA 基因高通量测序法检测家兔IF 丰度;以酶联免疫吸附测定法(ELISA) 行直肠、迷走神经、下丘脑和颈交感干中血管活性肠多肽(VIP)、神经肽Y(NPY) 的检测及斜方肌、C3~5 椎 间盘组织中脂多糖(LPS) 的检测。以Pearson 分析检测IF 丰度与相应各组织中VIP、NPY 及LPS 的相关性。 结果:最终纳入统计,空白组3 只,模型组3 只,桂枝加葛根汤组3 只。造模后,模型组与空白组厚壁菌门与 拟杆菌门的丰度,差异有统计学意义(P<0.05),提示家兔IF 失调已形成。给药后,中药组与模型组厚壁菌 门与拟杆菌门2 种菌群丰度分别比较,差异均有统计学意义(P<0.05),提示中药能有效改变菌群丰度,恢复 正常的厚壁菌/拟杆菌比例。与空白组比较,模型组VIP 和NPY 在直肠、迷走神经、下丘脑和颈交感神经干中 的含量均明显升高,LPS 在斜方肌和C3~5 椎间盘组织中含量亦明显升高,差异均有统计学意义(P<0.05);与 模型组比较,中药组上述各指标含量均明显降低(P<0.05)。各相应组织中VIP、NPY 及LPS 的水平变化与厚 壁菌群丰度呈正相关性(P<0.01),而与拟杆菌群丰度呈负相关性(P<0.05)。结论:IF 失调与NTCS 存在密 切相关性。在NTCS 发病过程中,IF 失调可通过菌-肠-脑-颈交感轴将促炎物质VIP、NPY 及LPS 等损伤信号 上传至颈部,从而促进颈部软组织诸如斜方肌和颈椎间盘组织内的免疫炎症反应。而桂枝加葛根汤则具有IF 调节作用,即通过恢复正常菌群比例,抑制有害信号在菌-肠-脑-颈交感轴内上传,从而消除颈周软组织的炎 症反应以达止痛效果。

    Abstract:

    Abstract:Objective:To study the mechanism of Guizhi and Gegen Decoction for neck- type cervical spondylopathy (NTCS) in rabbits based on intestinal flora (IF) regulation. Methods:A total of 9 rabbits were randomly divided into the blank group,the model group,and the Guizhi and Gegen Decoction group,with 3 rabbits in each group. The blank group was not subjected to modeling and the rest two groups were made as the NTCS rabbit models with spleen deficiency syndrome. The blank group and the model group were not given any medicine, and the Guizhi and Gegen Decoction group was treated with 10mL/kg Guizhi and Gegen Decoction by gavage for seven days,once per day. The IF abundance of rabbits was detected by 16 SrRNA gene high throughput sequencing; the vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY) of rectum,vagus nerve,hypothalamus and cervical sympathetic trunk,and the lipopolysaccharide (LPS) of C3- 5 intervertebral disc tissue were detected by enzyme linked immunosorbent assay (ELISA); The correlation of IF abundance with VIP,NPY and LPS of each tissue was detected by Pearson correlation analysis. Results: According to final statistics, 3 rabbits were included in the blank group, the model group and the Guizhi and Gegen Decoction group, respectively. After modeling, the abundance of Firmicutes and Bacteroidetes in the model group and the blank group showed significance in the difference (P<0.05), indicating that the IF imbalance in rabbits had been formed. After medication, comparing respectively the abundance of Firmicutes and Bacteroidetes between the Chinese medicine group and the model group, differences being significant (P<0.05), indicating that Chinese medicine can effectively change the abundance of flora and recover the normal proportion of Firmicutes and Bacteroidetes. The contents of VIP and NPY of rectum, vagus nerve, hypothalamus and cervical sympathetic trunk in the model group were significantly increased when compared with those in the blank group, and the LPS content of cucullaris and C3- 5 intervertebral disc tissue was also obviously increased, differences being significant (P<0.05);the above contents in the Chinese medicine group were significantly decreased when compared with those in the model group (P<0.05). The levels of VIP,NPY and LPS in the corresponding tissues were positively correlated with the abundance of Firmicutes (P<0.01), and negatively correlated with the abundance of Bacteroidetes (P<0.05). Conclusion:IF imbalance is closely related to NTCS. During the onset of NTCS, IF imbalance can deliver the damage signals such as pro- inflammatory substances VIP,NPY and LPS to the neck through the bacteria-gut-brain-neck sympathetic axis,thereby promoting the immune inflammatory responses in soft tissues of neck such as cucullaris and cervical intervertebral disc tissue. Guizhi and Gegen Decoction can regulate IF,that is,by restoring the normal proportion of flora,it inhibits the delivery of damage signals via the bacteria- gut- brain- neck sympathetic axis, thereby eliminating the inflammatory responses of the soft tissue around the neck to achieve analgesic effect.

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秦宇航.基于肠道菌群调节研究桂枝加葛根汤治疗颈型颈椎病家兔作用机制[J].新中医,2024,56(2):186-190

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  • 在线发布日期: 2024-01-25
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