Bushen Wenyang Huayu Decoction inhibits autophagy by regulating the SIRT1-FoXO-1 pathway in endometriosis rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Bushen Wenyang Huayu Decoction (BWHD) is a traditional Chinese medicine for tonifying kidney and warming Yang, thereby resolving blood stasis and relieving pain. BWHD can significantly improve the clinical symptoms of patients with endometriosis (EMs), but its mechanism is still unclear. AIM OF THE STUDY: We evaluated the expression and role of the SIRT1-FoxO-1 pathway and autophagy levels in EMs rats. The therapeutic effects and potential therapeutic mechanisms of BWHD were also investigated. METHODS: Twenty rats were randomized into the sham group and eighty rats were used for model establishment by autologous transplantation. After successful modeling, they were randomized into the model, BWHD, EX527+BWHD and EX527 groups, with 20 rats in each group. All rats were intragastrically administered with for 3 weeks. Localization of Sirtuin 1 (SIRT1), Forkhead boxO-1 (FoXO-1), Beclin-1, autophagy-related 5 (Atg5) and autophagy-related 7 (Atg7) was determined by immunohistochemical staining. The expression of the above proteins was determined by Western blot and their messenger RNA (mRNA) levels were detected by Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). RESULTS: The protein and mRNA expressions of FoXO-1, Beclin-1, Atg5 and Atg7 in the model group were markedly increased, while that of SIRT1 was markedly decreased relative to the sham group (p < 0.05 and p<0.01, respectively). Results showed that the protein and mRNA expressions of FoXO-1, Beclin-1, Atg5 and Atg7 in eutopic and ectopic endometrium of BWHD group were lower, while SIRT1 expression was higher than in the model group (p < 0.05 and p<0.01, respectively). Furthermore, protein and mRNA expression levels of FoXO-1, Beclin-1, Atg5 and Atg7 in eutopic and ectopic endometrium of EX527 group were higher, while SIRT1 level was significantly lower than in the model group (p < 0.05 and p < 0.01, respectively). The EX527-induced changes in protein and mRNA expressions were reversed in the EX527+BWHD group (p < 0.05 and p < 0.01, respectively). CONCLUSIONS: BWHD inhibits autophagy by up-regulating SIRT1 and down-regulating FoXO-1 expression in EMs via the SIRT1-FoXO-1 signaling pathway. Therefore, it is a potential treatment for EMs.

Bushen Wenyang Huayu Decoction Targets TLR4/NF-κB Mediated Autophagy to Treat Endometriosis Effectively.

Endometriosis has been found to be closely related to autophagy. This study aimed to elucidate the possible mechanism of Bushen Wenyang Huayu Decoction (BWHD) in treating endometriosis (EMs) by targeting TLR4/NF-κB-mediated autophagy. Autologous grafting was used to generate the EMs model in rats. Once the model was developed, BWHD high-dose and low-dose groups received intragastric administration of BWHD, and the gestrinone group served as a positive control. Immunofluorescence labeling and Western blotting were used for the protein expression of toll-like receptor 4 (TLR4), nuclear transcription factor-κB (NF-κB), Beclin-1, and selective autophagy connector protein P62 (P62). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to analyze mRNA levels of TLR4, NF-κB, Beclin-1, and P62. We found that BWHD significantly reduced the size of ectopic lesions in rats with EMs, regulated reproductive hormone levels, and alleviated the cell autophagy level. It suggested that BWHD could be an effective treatment of EMs by targeting TLR4/NF-κB signaling pathway.

[Effects of Bushen Wenyang Huayu Recipe on TRPV1 and Sensitization Factor NGF in Experimental Endometriosis].

OBJECTIVE: To study the effect of Bushen Wenyang Huayu Recipe (BSWYHYR) on nerve growth factor (NGF) and transient receptor potential vanilloid receptor I (TRPV1) in experimental endometriosis (EMT), and to explore its mechanism for treating EMT-induced pain. METHODS: Totally in-bred line BALB/c 75 female mice were divided into five groups, i.e., the sham-operation group, the model group, the high dose BSWYHYR group, the low dose BSWYHYR group, the gestrinone group, 15 in each group. Writhing response was observed in each group. Serum contents of NGF were detected using ELISA. Expression levels of NGF and TRPV1 in uterus and ectopic foci were detected using immunohistochemical staining SP and Western blot. mRNA expression levels of NGF and TRPV1 in uterus and ectopic foci were detected by Real-time PCR. RESULTS: The serum NGF content in the model group was higher than that in the sham-operation group (P < 0.01), and there was positive correlation between NGF and the writhing frequency (r = 0.574, P < 0.01). Compared with the model group, serum levels of NGF significantly decreased in the 3 treatment groups (P < 0.05). Compared with the sham-operation group, mRNA and protein expression levels of NGF and TRPV1 increased significantly in the model group (P < 0.01). Protein expression levels of NGF and TRPV1 decreased significantly in the 3 treatment groups, when compared with the model group (P < 0.01). mRNA expression levels of NGF and TRPV1 decreased most in the high dose BSWYHYR group (P < 0.01). NGF in uterus and ectopic foci was positively correlated with protein and mRNA expression levels of TRPV1 (P < 0.01). CONCLUSIONS: NGF and TRPV1 participated in the occurrence of pain in EMS. BSWYHYR played an important role in inhibiting EMT-induced pain through reducing the up-regulation of NGF on TRPV1.

Spatial Heterogeneity of Soil Nutrients after the Establishment of Caragana intermedia Plantation on Sand Dunes in Alpine Sandy Land of the Tibet Plateau.

The Gonghe Basin region of the Tibet Plateau is severely affected by desertification. Compared with other desertified land, the main features of this region is windy, cold and short growing season, resulting in relatively difficult for vegetation restoration. In this harsh environment, identification the spatial distribution of soil nutrients and analysis its impact factors after vegetation establishment will be helpful for understanding the ecological relationship between soil and environment. Therefore, in this study, the 12-year-old C. intermedia plantation on sand dunes was selected as the experimental site. Soil samples were collected under and between shrubs on the windward slopes, dune tops and leeward slopes with different soil depth. Then analyzed soil organic matter (SOM), total nitrogen (TN), total phosphorus (TP), total potassium (TK), available nitrogen (AN), available phosphorus (AP) and available potassium (AK). The results showed that the spatial heterogeneity of soil nutrients was existed in C. intermedia plantation on sand dunes. (1) Depth was the most important impact factor, soil nutrients were decreased with greater soil depth. One of the possible reasons is that windblown fine materials and litters were accumulated on surface soil, when they were decomposed, more nutrients were aggregated on surface soil. (2) Topography also affected the distribution of soil nutrients, more soil nutrients distributed on windward slopes. The herbaceous coverage were higher and C. intermedia ground diameter were larger on windward slopes, both of them probably related to the high soil nutrients level for windward slopes. (3) Soil "fertile islands" were formed, and the "fertile islands" were more marked on lower soil nutrients level topography positions, while it decreased towards higher soil nutrients level topography positions. The enrichment ratio (E) for TN and AN were higher than other nutrients, most likely because C. intermedia is a leguminous shrub.

[Water physiological characteristics and leaf traits of different aged Salix cheilophila on alpine sandy land].

Taking 4-, 11-, 25-, and 37- year old Salix cheilophila stands on the alpine sandy land of Gonghe basin in Qinghai of West China as test objects, a laboratory test was conducted on their relative water deficit, water holding ability, specific leaf area (SLA), leaf mass-based nitrogen concentration (N(mass)) and phosphorous concentration (P(mass)), and N(mass)/P(mass), aimed to understand the variation patterns of the water physiological characteristics and leaf traits of different aged S. cheilophila on alpine sandy land. No significant difference was observed in the relative water deficit of the four stands. The daily mean value of water potential of the 37-year old stand was significantly lower, as compared with that of the other three stands, and the 4- and 11-year old stands had a significantly lower daily mean water potential than the 25-year old stand. The water loss rate of the 4-year old stand was significantly lower than that of the other three stands, and the 25-year old S. cheilophila stand had a significantly lower water loss rate than the 11-year old stand. The 4-year old stand also had a significantly lower SLA than the other three stands, implying its higher water use efficiency. The N(mass) of the 11-year old stand was significantly higher than that of the other three stands, and the 25-year old stand had a significantly higher N(mass) than the 37-year old stand, implying that the 11- and 25-year old stands had a higher photosynthetic capacity. The P(mass) of the 11-year old stand was significantly higher than that of the 25- and 37-year old stands, and the 4-year old stand had a significantly higher P(mass) than the 25-year old stand. The N(mass)/P(mass) of the four stands was 5.16-6.28, and the 25-year old stand had a significantly higher N(mass)/P(mass) than the 4- and 11-year old stands. The N(mass) of the four stands was significantly positively correlated with P(mass) the P(mass) was highly significantly negatively correlated with N(mass)/P(mass) and significantly nega- tively correlated with stand age, and the N(mass)/P(mass) was significantly positively correlated with stand age. It was suggested that S. cheilophila at its different developmental stages could have different ecological adaptive strategies.