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电针联合壮医药线点灸对糖尿病胃轻瘫大鼠胃窦沉默信息调节因子-1/核因子κB信号通路的影响

[Effect of electroacupuncture combined with Zhuang-medicine-thread moxibustion on silent information regulator-1/nuclear factor κB signaling pathway in gastric antrum of diabetic gastroparesis rats].

作者信息

Mai Wei, Fan Yu-Shan, Miao Fu-Rui, He Cai, Huang Li-Lin, Zhao Xiao-Jun, Nie Ying-Jie, Pang Rui-Kang

机构信息

Graduate School, Guangxi University of Chinese Medicine, Nanning 530001, China.

College of Acupuncture-moxibustion and Tuina, Guangxi University of Chinese Medicine, Nanning 530001, China.

出版信息

Zhen Ci Yan Jiu. 2021 Oct 25;46(10):837-44. doi: 10.13702/j.1000-0607.200942.

DOI:10.13702/j.1000-0607.200942
PMID:34698457
Abstract

OBJECTIVE

To observe the effect of electroacupuncture (EA) combined with Zhuang-medicine-thread moxibustion on silent information regulator-1 (SIRT1)/nuclear factor kappa B (NF-κB) signal pathway and inflammatory factor expression in gastric antrum tissue of diabetic gastroparesis (DGP) rats, so as to explore its mechanism underlying improvement of DGP.

METHODS

Male SD rats were randomly divided into normal, model, medication, EA, Zhuang-medicine-thread moxibustion (moxibustion) and EA+moxibustion groups (=12 per group). The DGP model was established by intraperitoneal injection of streptozotocin (STZ). Rats of the medication group were treated by gavage of 0.15 mg/mL mosapride citrate suspension. EA (10 Hz /50 Hz, 2 mA) or moxibustion (3 cones) or EA+moxibustion was applied to "Zhongwan"(CV12), bilateral "Neiguan"(PC6) and bilateral "Sanyinjiao"(SP6) of the related group for 20 min, once a day for 3 weeks. Blood glucose, gastric emptying rate and intestinal propulsion rate were measured. The levels of serum interleukin (IL)-6, IL-8, IL-10 and tumor necrosis factor-α(TNF-α) were detected by ELISA; the phosphorylation level of the phosphorylated inhibitor of nuclear factor κBα inhibitor (pIκ-Bα), the protein and mRNA expression of NF-κB p65 and SIRT1 in the gastric antrum tissue were detected by Western blot and real-time quantifitative PCR, respectively.

RESULTS

(1) Compared with the normal group, the levels of blood glucose, serum IL-6, IL-8, TNF-α, and gastric pIκ-Bα and NF-κB p65 protein and mRNA expressions were significantly increased (<0.01), and the gastric emptying rate, intestinal propulsion rate, serum IL-10 level, and SIRT1 protein and mRNA expressions were considerably decreased in the model group (<0.01). (2) In contrast to the model group, the blood glucose in the EA, moxibustion and EA+moxibustion groups, serum IL-6, IL-8 and TNF-α levels in the 4 treatment groups, as well as NF-κB p65 protein expression in the medication and EA+moxibustion groups, and NF-κB p65 mRNA expression and pIκ-Bα protein and mRNA expression in the 4 treatment groups were significantly decreased (<0.01, <0.05); while the gastric emptying rate and intestinal propulsive rate and IL-10 content in the 4 treatment groups, and SIRT1 protein and mRNA expression in the medication and EA+moxibustion groups were obviously increased (<0.05, <0.01). (3) The effects of EA+moxibustion were significantly superior to those of simple EA and moxibustion in increasing gastric emptying rate, IL-10, SIRT1 protein expression (<0.05, <0.01), and in lowering IL-8 and TNF-α contents, pIκ-Bα protein and mRNA expression and NF-κB p65 mRNA expression (<0.05, <0.01). No significant differences were found among the 4 intervention groups in promoting the intestinal propulsive rate and among the EA, moxibustion and EA+moxibustion groups in lowering blood glucose (>0.05).

CONCLUSION

EA combined with Zhuang-medicine-thread moxibustion can effectively reduce the level of serum inflammatory factors and regulate SIRT1/NF-κB signal pathway in DGP rats, which may contribute to its function in improving gastrointestinal movement.

摘要

目的

观察电针(EA)联合壮医药线点灸对糖尿病胃轻瘫(DGP)大鼠胃窦组织中沉默信息调节因子1(SIRT1)/核因子κB(NF-κB)信号通路及炎症因子表达的影响,以探讨其改善DGP的作用机制。

方法

将雄性SD大鼠随机分为正常组、模型组、药物组、电针组、壮医药线点灸组(灸疗组)和电针+灸疗组(每组n = 12)。采用腹腔注射链脲佐菌素(STZ)建立DGP模型。药物组大鼠灌胃给予0.15 mg/mL枸橼酸莫沙必利混悬液。电针组(10 Hz/50 Hz,2 mA)、灸疗组(3壮)或电针+灸疗组对相关穴位“中脘”(CV12)、双侧“内关”(PC6)和双侧“三阴交”(SP6)进行干预20 min,每天1次,共3周。检测血糖、胃排空率和肠推进率。采用酶联免疫吸附测定(ELISA)法检测血清白细胞介素(IL)-6、IL-8、IL-10和肿瘤坏死因子-α(TNF-α)水平;采用蛋白质免疫印迹法(Western blot)和实时定量聚合酶链反应(real-time quantifitative PCR)分别检测胃窦组织中核因子κBα抑制蛋白(pIκ-Bα)的磷酸化水平、NF-κB p65和SIRT1的蛋白及mRNA表达。

结果

(1)与正常组比较,模型组大鼠血糖、血清IL-6、IL-8、TNF-α水平及胃pIκ-Bα、NF-κB p65蛋白及mRNA表达均显著升高(P<0.01),胃排空率、肠推进率、血清IL-10水平及SIRT1蛋白和mRNA表达均显著降低(P<0.01)。(2)与模型组比较,电针组、灸疗组和电针+灸疗组大鼠血糖水平,4个治疗组血清IL-6、IL-8、TNF-α水平,药物组和电针+灸疗组NF-κB p65蛋白表达,4个治疗组NF-κB p65 mRNA表达及pIκ-Bα蛋白和mRNA表达均显著降低(P<0.01,P<0.05);4个治疗组胃排空率、肠推进率及IL-10含量,药物组和电针+灸疗组SIRT1蛋白和mRNA表达均明显升高(P<0.05,P<0.01)。(3)电针+灸疗组在提高胃排空率、IL-10、SIRT1蛋白表达(P<0.05,P<0.01),降低IL-8、TNF-α含量、pIκ-Bα蛋白和mRNA表达及NF-κB p65 mRNA表达(P<0.05,P<0.01)方面,效果明显优于单纯电针组和灸疗组。4个干预组在促进肠推进率方面差异无统计学意义;电针组、灸疗组和电针+灸疗组在降低血糖方面差异无统计学意义(P>0.05)。

结论

电针联合壮医药线点灸能有效降低DGP大鼠血清炎症因子水平,调节SIRT1/NF-κB信号通路,这可能是其改善胃肠运动功能的作用机制。

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