神经固本方调控PI3K和NRF2信号通路治疗免疫性不孕的机制研究

Study on the Mechanism of Shenjing Guben Prescription Regulating PI3K and NRF2 Signaling Pathway in the Treatment of Immune Infertility.

作者信息

Liu Handu, Xue Jianguo, Mo Hui

机构信息

Faculty of Chinese Medicine, Macau University of Science and Technology, Wai Long Avenue, S.A.R. 999078, Taipa, Macau.

Andrology, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China.

出版信息

Evid Based Complement Alternat Med. 2022 May 13;2022:8754188. doi: 10.1155/2022/8754188. eCollection 2022.

DOI:10.1155/2022/8754188

PMID:35600964

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9122677/

Abstract

OBJECTIVE

To explore the mechanism of Shenjing Guben prescription (SP) in the treatment of immune infertility by regulating PI3K-NRF2/p38 signal pathway.

METHODS

60 adult male SD rats were randomly divided into control group (NC group), ACN group, low concentration AP intervention group (low group), middle concentration SP intervention group (middle group), and high concentration SP intervention group (high group). 12 rats in each group were administered by gavage once a day, 6 days/w, and the rats were killed after 28 days. Bilateral testis and epididymis were removed and weighed and organ coefficients were calculated, and testicular histopathological sections were prepared to evaluate the changes of testicular tissue structure. The relative expression levels of PI3K, MKK7, JNK, p38 mRNA, and protein in testis were measured by QRT-PCR and western blot.

RESULTS

(1) Compared with the control group, the proportion of grade A and B sperms in ACN group increased significantly, and the proportion of grade D sperm decreased significantly ( < 0.05). After SP intervention, compared with ACN group, there was no significant difference in the proportion of sperm at all levels in low, medium, and high SP intervention groups ( > 0.05). (2) Compared with the control group, the sperm VCL, VSL, VAP, and mad in ACN group increased significantly, and the BCF decreased significantly ( < 0.05). After SP intervention, compared with ACN group, there was no significant difference in sperm motility parameters among low, medium, and high SP intervention groups ( > 0.05). (3) Compared with the control group, the activities of AKP and SDH in testicular tissue of rats in ACN group decreased significantly ( < 0.05). After SP intervention, compared with ACN group, AKP activity increased significantly and LDH activity decreased significantly in low, medium, and high SP intervention groups ( < 0.05). (4) Compared with the control group, the expression levels of PI3K, p-PI3K, MKK7, p-MKK7, JNK, p-JNK, p38, and p-p38 proteins and the ratios of p-JNK/JNK and p-p38/p38 increased in the testis of ACN group ( < 0.05). After SP intervention, compared with ACN group, the protein expression levels of PI3K, p-PI3K, MKK7, p-MKK7, JNK, p-JNK, p38, and p-p38 in testicular tissue of SP intervention group decreased, and the ratio of p-JNK/JNK and p-p38/p38 decreased ( < 0.05).

CONCLUSION

SP can reduce the oxidative stress of testis induced by ACN and inhibit the activation of PI3K-NRF2/p38 signal pathway.

摘要

目的

探讨神经固本方（SP）通过调节PI3K-NRF2/p38信号通路治疗免疫性不育的机制。

方法

将60只成年雄性SD大鼠随机分为对照组（NC组）、醋酸棉酚组（ACN组）、低浓度SP干预组（低剂量组）、中浓度SP干预组（中剂量组）和高浓度SP干预组（高剂量组）。每组12只大鼠，每天灌胃给药1次，每周6天，28天后处死大鼠。取出双侧睾丸和附睾称重并计算脏器系数，制备睾丸组织病理切片，评估睾丸组织结构变化。采用QRT-PCR和western blot检测睾丸中PI3K、MKK7、JNK、p38 mRNA及蛋白的相对表达水平。

结果

（1）与对照组相比，ACN组A级和B级精子比例显著升高，D级精子比例显著降低（P<0.05）。SP干预后，低、中、高剂量SP干预组各级精子比例与ACN组相比，差异无统计学意义（P>0.05）。（2）与对照组相比，ACN组精子VCL、VSL、VAP和mad显著升高，BCF显著降低（P<0.05）。SP干预后，低、中、高剂量SP干预组精子运动参数与ACN组相比，差异无统计学意义（P>0.05）。（3）与对照组相比，ACN组大鼠睾丸组织中AKP和SDH活性显著降低（P<0.05）。SP干预后，低、中、高剂量SP干预组与ACN组相比，AKP活性显著升高，LDH活性显著降低（P<0.05）。（4）与对照组相比，ACN组睾丸组织中PI3K、p-PI3K、MKK7、p-MKK7、JNK、p-JNK、p38和p-p38蛋白表达水平及p-JNK/JNK和p-p38/p38比值升高（P<0.05）。SP干预后，SP干预组睾丸组织中PI3K、p-PI3K、MKK7、p-MKK7、JNK、p-JNK、p38和p-p38蛋白表达水平降低，p-JNK/JNK和p-p38/p38比值降低（P<0.05）。