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基于氧化应激探讨针刺“秩边”(BL 54)透“水道”(ST 28)对卵巢早衰大鼠凋亡相关因子表达的影响

[Effect of acupuncture at "Zhibian" (BL 54) through "Shuidao" (ST 28) on the expression of apoptosis-related factors in rats with premature ovarian insufficiency based on oxidative stress].

作者信息

Yan Jing, Zhao Ji-Yu, Yin Lu-Yun, Yan Xiao-Qin, Jin Xiao-Fei

机构信息

Second Clinical College of Shanxi University of CM, Jinzhong 030619, China.

Jinzhong Hospital of TCM Affiliated to Shanxi University of CM.

出版信息

Zhongguo Zhen Jiu. 2023 Apr 12;43(4):454-60. doi: 10.13703/j.0255-2930.20220705-k0001.

Abstract

OBJECTIVE

To explore the possible mechanism of acupuncture at "Zhibian" (BL 54) through "Shuidao" (ST 28) on premature ovarian insufficiency (POI) from the perspective of oxidative stress.

METHODS

Sixty female SD rats were randomly divided into a blank group, a model group, a sham acupuncture group, a medication group, and an acupuncture group, 12 rats in each group. Except the blank group, the rats in the remaining groups were intraperitoneally injected with cyclophosphamide to establish the POI model. After the model was successfully established, the rats in the acupuncture group were treated with acupuncture at "Zhibian" (BL 54) through "Shuidao" (ST 28), with a depth of about 12 mm, and the needle was retained for 30 min; the acupuncture was given once a day, for a total of 4 weeks. The rats in the sham acupuncture group were treated with blunt-head needle to tap the skin surface of "Zhibian" (BL 54), without penetrating the skin, once a day for 4 weeks. The rats in the medication group were treated with estradiol valerate by gastric gavage for 4 weeks. After the intervention, the level of reactive oxygen species (ROS) in the ovarian tissue was detected by fluorescence probe; the expression of c-Jun N-terminal kinase (JNK), forkhead box O1 (FoxO1), tumor suppressor gene protein 53 (p53) and p53 up-regulated modulator of apoptosis (Puma) mRNA and protein in ovarian tissue were detected by real-time fluorescence quantitative PCR and Western blot.

RESULTS

Compared with the blank group, the level of ROS and the expression of JNK mRNA, p-JNK protein, FoxO1, p53, Puma mRNA and protein in the ovarian tissue in the model group were increased (<0.01). Compared with the model group, the level of ROS and the expression of p-JNK protein, FoxO1, p53, Puma mRNA and protein in the ovarian tissue in the sham acupuncture group were slightly reduced, but the difference was not statistically significant (>0.05). The level of ROS and the expression of JNK mRNA, p-JNK protein, FoxO1, p53, Puma mRNA and protein in the ovarian tissue in the acupuncture group and the medication group were reduced (<0.01).

CONCLUSION

Acupuncture at "Zhibian" (BL 54) through "Shuidao" (ST 28) could improve the level of oxidative stress, down-regulate the expression of apoptosis-related factors JNK, FoxO1, p53 and Puma induced by oxidative stress, and inhibit the premature failure of ovarian reserve function caused by apoptosis of ovarian granulosa cells in POI rats.

摘要

目的

从氧化应激角度探讨针刺“秩边”(BL 54)透“水道”(ST 28)对卵巢早衰(POI)的可能作用机制。

方法

将60只雌性SD大鼠随机分为空白组、模型组、假针刺组、药物组和针刺组,每组12只。除空白组外,其余各组大鼠腹腔注射环磷酰胺建立POI模型。模型成功建立后,针刺组大鼠采用针刺“秩边”(BL 54)透“水道”(ST 28)治疗,进针深度约12 mm,留针30 min;每天针刺1次,共4周。假针刺组大鼠用钝头针轻叩“秩边”(BL 54)皮肤表面,不穿透皮肤,每天1次,共4周。药物组大鼠采用戊酸雌二醇灌胃治疗4周。干预后,用荧光探针检测卵巢组织中活性氧(ROS)水平;采用实时荧光定量PCR和蛋白质印迹法检测卵巢组织中c-Jun氨基末端激酶(JNK)、叉头框O1(FoxO1)、肿瘤抑制基因蛋白53(p53)和p53上调凋亡调节因子(Puma)mRNA及蛋白表达。

结果

与空白组比较,模型组卵巢组织中ROS水平及JNK mRNA、p-JNK蛋白、FoxO1、p53、Puma mRNA和蛋白表达升高(<0.01)。与模型组比较,假针刺组卵巢组织中ROS水平及p-JNK蛋白、FoxO1、p53、Puma mRNA和蛋白表达略有降低,但差异无统计学意义(>0.05)。针刺组和药物组卵巢组织中ROS水平及JNK mRNA、p-JNK蛋白、FoxO1、p53、Puma mRNA和蛋白表达降低(<0.01)。

结论

针刺“秩边”(BL 54)透“水道”(ST 28)可改善氧化应激水平,下调氧化应激诱导的凋亡相关因子JNK、FoxO1、p53和Puma表达,抑制POI大鼠卵巢颗粒细胞凋亡所致的卵巢储备功能过早衰竭。

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