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.

Corrigendum to "Berberine Reduces Renal Cell Pyroptosis in Golden Hamsters with Diabetic Nephropathy through the Nrf2-NLRP3-Caspase-1-GSDMD Pathway".

[This corrects the article DOI: 10.1155/2021/5545193.].

Role and Mechanism of the Renin-Angiotensin-Aldosterone System in the Onset and Development of Cardiorenal Syndrome.

Cardiorenal syndrome (CRS), a clinical syndrome involving multiple pathological mechanisms, exhibits high morbidity and mortality. According to the primary activity of the disease, CRS can be divided into cardiorenal syndrome (type I and type II), renal heart syndrome (type III and type IV), and secondary heart and kidney disease (type V). The renin-angiotensin-aldosterone system (RAAS) is an important humoral regulatory system of the body that exists widely in various tissues and organs. As a compensatory mechanism, the RAAS is typically activated to participate in the regulation of target organ function. RAAS activation plays a key role in the pathogenesis of CRS. The RAAS induces the onset and development of CRS by mediating oxidative stress, uremic toxin overload, and asymmetric dimethylarginine production. Research on the mechanism of RAAS-induced CRS can provide multiple intervention methods that are of great significance for reducing end-stage organ damage and further improving the quality of life of patients with CRS.

Berberine Reduces Renal Cell Pyroptosis in Golden Hamsters with Diabetic Nephropathy through the Nrf2-NLRP3-Caspase-1-GSDMD Pathway.

Objective: To observe the effect of berberine (BBR) on kidney cell pyroptosis in golden hamsters with diabetic nephropathy (DN) and to explore the molecular mechanism of its renal protection. Methods: Fifty clean-grade male golden hamsters were randomly divided into a control group (10) and a model building group (40). The DN model was established by high-sugar and high-fat feeding and injection of a small amount of STZ. After successful establishment of the model, they were randomly divided into a model group, western medicine group, and berberine high- and low-dose groups. The western medicine group was given irbesartan 13.5 mg/kg, and the berberine high- and low-dose groups were given BBR 200 mg/kg and 100 mg/kg, respectively, for 8 consecutive weeks. An automatic biochemical analyser was used to measure blood glucose, blood lipids, kidney function, MDA, and other indicators; radioimmunoassay was used to assess serum insulin; enzyme-linked immunosorbent assay (ELISA) was used to quantify IL-1ß, IL-6, IL-18, TNF-α; HE, PAS, and Masson staining were used to observe kidney pathological tissue morphology; western blot and real-time fluorescent quantitative PCR were used to assess protein and mRNA expression of molecules, such as Nrf2, NLRP3, Caspase-1, and GSDMD; and TUNEL staining was used to detect DNA damage. SPSS statistical software was used for the data analysis. Results: The kidney tissues of golden hamsters in the control group were normal; Nrf2 was highly expressed, serum MDA level was low, NLRP3 expression in kidney tissue was not obvious, Caspase-1 and GSDMD were weakly expressed, and only a few TUNEL-positive cells were observed. Compared with the control group, the golden hamsters in the model group had obvious renal pathological damage; blood glucose, blood lipids, renal function-related indexes, insulin, and inflammatory factors IL-1ß, IL-6, IL-18, and TNF-α were increased (P < 0.05); NLRP3, Caspase-1, and GSDMD expression was increased; Nrf2 expression was decreased; MDA level was increased (P < 0.05); and the number of TUNEL-positive cells was increased. Compared with the model group, the pathological morphology of the kidney tissue of golden hamsters in the three treatment groups was significantly improved; blood glucose, blood lipids, renal function, and the expression of inflammatory factors IL-1ß and IL-6 were reduced (P < 0.05); NLRP3, Caspase-1, GSDMD, and other molecular proteins and mRNA expression were decreased; Nrf2 expression was increased; MDA level was decreased (P < 0.05); and the number of TUNEL-positive cells was decreased. Conclusion: DN golden hamster kidney NLRP3-Caspase-1-GSDMD signalling was enhanced. BBR can reduce oxidative stress damage by regulating antioxidative Nrf2 and then regulating NLRP3-Caspase-1-GSDMD signalling to inhibit pyroptosis, antagonizing DN inflammation-induced damage.

[Effect of GnRHa controlled ovarian hyperstimulation on mouse embryo implantation and its mechanism].

The purpose of the present study was to investigate the effects and underlying mechanism of gonadotropin-releasing hormone agonist (GnRHa) controlled ovarian hyperstimulation (COH) on embryo implantation in mice. Forty female Kunming mice aged 9 weeks were randomly divided into two groups (control and COH groups). The COH group received intraperitoneal (i.p.) injections of aminocyclin acetate (GnRHa), human menopausal gonadotropin (HMG) and human chorionic gonadotropin (hCG), while the control group was given equal amount of physiological saline by i.p. injection. One male mouse and two female mice were put into the same cage at 16:00 on the hCG injection day, and on the fourth day of pregnancy, 10 mice from each group were killed. The levels of serum estradiol (E2) and progesterone (P) were measured by radioimmunoassay; HE staining was used to observe the morphology of ovarian and endometrial tissues. The protein expression levels of endometrial leukemia inhibitory factor (LIF), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), heparin-binding epidermal growth factor-like growth factor (HB-EGF) and glycodelin A were detected by Western blot and immunohistochemistry. Ten mice from each group were sacrificed on the eighth day of pregnancy, and the status of the uterus and the average number of blastocysts were observed. The results showed that, compared with control group, the serum E2 level in COH group was significantly decreased (P < 0.05), while the P level was increased significantly (P < 0.05); the ovarian follicles at different developmental stages were rare, corpus lutea (CL) were visible and multiple, the endometrium was thinned, and the number of endometrial glands was reduced (P < 0.05); the contents of LIF, p-STAT3, HB-EGF and glycodelin A in the endometrium were decreased significantly (P < 0.05) on the fourth day of pregnancy; mouse blastocysts developed slowly and were decreased in number on the eighth day of pregnancy (P < 0.05). The above results suggest that GnRHa COH can affect embryo implantation in mice. The mechanism may be related to the imbalance of gonadal hormone, the changes in the structure of the endometrium and the expressions of LIF, p-STAT3, HB-EGF and glycodelin A in the implantation stage, which may lead to the decrease of endometrial receptivity and the abnormal dialogue between the embryo and the uterus.

[Effect of Bushen Tiaojing Recipe containing serum on FSH/cAMP-PKA pathway in in vitro cultured human ovarian granular cells].

OBJECTIVE: To explore the potential molecular mechanisms for Bushen Tiaojing Recipe (BTR) improving the endocrine function of ovarian granular cells by observing the effect of BTR containing serum on follicle stimulating hormone/cyclic adenosine monophosphate-protein kinase A (FSH/ cAMP-PKA) pathway in in vitro cultured human ovarian granular cells. METHODS: The primary ovarian granular cells collected from in vitro fertilization-embryo transfer patients were cultured for 24 h. The human and rat serum containing different concentrations of BTR (low, medium, high dose), and their normal serums were co-incubated with ovarian granular cells for 48 h respectively, and then they were divided into the low, medium, high dose BTR groups and the control group. The levels of estradiol (E2), progesterone (P), and cyclic adenosine monophosphate (cAMP) in the culture medium were measured by radioimmunoassay. The protein expression of FSHR in ovarian granular cells was detected by Western Blot. The mRNA expression of follicle stimulating hormone receptor (FSHR) and P450 aromatase (P450arom) in ovarian granular cells were detected by Real-time PCR. RESULTS: In human BTR containing serum groups: Compared with control group, the levels of E2 and cAMP in the culture medium were higher (both P < 0.05) in the medium and high dose BTR groups; the levels of P in the culture medium decreased in the medium dose BTR group (P < 0.01). The protein and mRNA expression of FSHR in ovarian granular cells increased (all P < 0.01), the mRNA expressions of P450arom in ovarian granular cells were higher (P < 0.05, P< 0.01) in the medium and high dose BTR groups. In rat BTR containing serum groups: Compared with the control group, the levels of E2 in the culture medium were higher (all P < 0.01), cAMP in the culture medium were higher (P < 0.05, P < 0.01) in the medium and high dose BTR group; the levels of P in the culture medium decreased in the medium dose BTR group (P < 0.01). The protein and mRNA expression of FSHR in ovarian granular cells were higher (all P < 0.01), the mRNA expression of P450arom in ovarian granular cells increased in the medium and high dose BTR groups (P < 0.05, P < 0.01). CONCLUSION: BTR could possibly improve the endocrine function of ovarian granular cells by regulating main effector molecules FSHR, cAMP, P450arom, and E2 in FSH/cAMP-PKA pathway of ovarian granular cells.

Effect of electroacupuncture on brain-derived neurotrophic factor mRNA expression in mouse hippocampus following cerebral ischemia-reperfusion injury.

OBJECTIVE: This work aims to observe the effects of electroacupuncture on brain-derived neurotrophic factor (BDNF) mRNA expression in mouse hippocampus following cerebral ischemia-reperfusion injury. METHODS: The models of mouse cerebral ischemia-reperfusion injury were established. A total of 96 healthy mice were randomly assigned into 4 groups, namely, the sham surgery, model, model + electroacupuncture, and mode + hydergine groups. Mice in the model + electroacupuncture group were treated through electroacupuncture at the Shenshu (BL 23), Geshu (BL 17), and Baihui (GV 20) acupoints. Mice in the model+hydergine group were intragastrically administered with hydergine (0.77 mg/kg(-1) x day(-1)). The levels of BDNF mRNA expressions in the hippocampus were ana lyzed through a semi-quantitative reverse transcription-polymerase chain reaction assay on days 1 and 7 after the surgeries. RESULTS: BDNF mRNA expressions in the mouse hippocampus of the model group on days 1 and 7 after the surgery were higher than those of the sham surgery group (both P < 0.01). On days 1 and 7 of the electroacupuncture treatment, BDNF mRNA expression in the mouse hippocampus of the model + electroacupuncture group was significantly elevated compared with the model group (both P < 0.01) or the model + hydergine group (both P < 0.01). On days 1 and 7 of the hydergine treatment, BDNF mRNA expression in the mouse hippocampus of the model + hydergine group tended to increase compared with the model group; however, statistical significance was not achieved (both P > 0.05). CONCLUSION: Electroacupuncture treatment enhances endogenous BDNF expression, which may improve the survival environment for intracerebral neurons and inhibit the apoptosis of hippocampal cells.

[Effects of buzhong yiqi decoction on adriamycin induced heart failure in rats].

OBJECTIVE: To study the effects and mechanism of Buzhong Yiqi decoction on adriamycin-induced acute myocardial injury in rats. METHOD: 50 rats were randomly divided to five groups: control group, heart failure group, low dose Buzhong Yiqi decoction, high dose Buzhong Yiqi decoction and captopril group. Adriamycin was injected into the latter four groups to built a model of heart failure. Then, the effects of different doses of Buzhong Yiqi decoction on hemodynamics, cardiac tissue histological changes, antioxidant capacity and apoptosis of the damaged hearts were studied. RESULT: Adriamycin led to myocardial fiber swelling and fracture, Buzhong Yiqi decoction could reduce myocardial lesions. Buzhong Yiqi decoction could also improve heart antioxidant capacity and inhibit adriamycin-induced cardiomyocyte apoptosis. CONCLUSION: Buzhong Yiqi decoction could significantly ease adriamycin induced heart failure in rats, and the mechanism is related to anti-oxidation and inhibiting apoptosis.

[Effect of high-frequency electroacupuncture on lipid metabolism in obesity rats].

OBJECTIVE: To observe the effect of high-frequency electroacupuncture (EA) on lipid metabolism in obesity rats. METHODS: Fifty-one SD rats were randomized into control group (n=13), model group (n=19) and EA group (n=19). Obesity model was duplicated by feeding the rats with high fat forage and subcutaneous injection of 15% glutamate sodium solution. EA (100 Hz, 3-5 mA) was applied to "Housanli" (ST 36), "Sanyinjiao" (SP 6), "Guanyuan" (CV 4) and "Zhongwan" (CV 12) for 10 min everyday and for 28 days. Lee's index, body weight, area and the number of the adipose cells around the left kidney were detected. Blood samples were collected for assaying serum triglyceride (TG), total cholesterol (TC) with cholesterol oxidase phenol 4-aminoantipyrine peroxidase (CHOD-PAP) method, glycerol phosphate oxidase-p-aminophenazone (GPO-PAP) methods, high density lipid-cholesterol (HDL-C) with phosphotungstic acid- Mg2+ method, low density lipid-cholesterol (LDL-C) with polyvingel sulfate (PVS) sedimentation method and plasma lipoprotein lipase (LPL) with immunoturbidmetry endpoint method, and serum insulin and leptin contents with radioimmunoassay respectively. RESULTS: After modeling, in comparison with control group, Lee's index, serum TG, TC, LDL-C, fat weight, adipose cell area, serum leptin and insulin contents in model group increased considerably (P<0.05, 0.01), while serum HDL-C, plasma LPL, adipose cell number decreased significantly (P<0.05, 0.01). Following EA, compared with model group, Lee's index, serum TG, TC, LDL-C, fat weight, adipose cell area, serum leptin and insulin levels decreased significantly in EA group (P<0.05, 0.01), whereas serum HDL-C, plasma LPL, and adipose cell number increased apparently (P<0.01). CONCLUSION: High frequency EA can effectively improve abnormal lipid metabolism, and reduce fat accumulation in obesity rats, which may be closely associated with its effect in regulating the balance between leptin and insulin levels.

[Effect of electroacupuncture on metabolism of lipids in rats of obesity induced by sodium glutamate].

OBJECTIVE: To explore mechanisms of acupuncture for reducing of body weight. METHODS: The rat obesity model was developed by subcutaneous injection of sodium glutamate solution, and the experimental rats were divided into a blank control group, a model group, an electroacupuncture group and a sibutramine group; blood TC, TG, HDL-C, LDL-C contents and lipoprotein lipase (LPL) activity, and serum leptin and insulin levels were determined. RESULTS: The body weight and Lee's index, and TG, TC and LDL-C contents in the electroacupuncture group decreased significantly as compared with those in the model group (all P<0.01), the actions in the electroacupuncture group being superior to those in the sibutramine group; the HDL-C contents in both the electroacupuncture and the sibutramine groups were higher than that of the model group (P<0.01) with no significant difference between the electroacupuncture group and the sibutramine group; LPL activity in the electroacupuncture group increased significantly as compared with that in the model group (P<0.01); serum leptin and insulin levels in both the electroacupuncture group and the sibutramine group were lower than that in the model group (P<0.05, P<0.01), with no significant difference between the electroacupuncture group and the sibutramine group in the effect on insulin content, and the decrease of leptin level in the electroacupuncture group was more as compared with the sibutramine group (P<0.01). CONCLUSION: Electroacupuncture can improve the status of high blood lipids, increase lipoprotein lipase activity, and regulate serum leptin and insulin levels in the obestic rat.

[Effect of Shenluotong on secretion of ECM and expression of TGF-beta1 in rats mesangial cells of rats].

OBJECTIVE: To investigate the effect of Shenluotong on the expression of transforming growth factor-beta1 (TGF-beta1) and extracellular matrix (ECM) in Ang II-induced MCs. METHOD: Fibronectin (FN) and collagen type IV (Col IV) of extracellular matrix were detected by enzyme-linked immunosorbent assay; the expression of TGF-beta1 mRNA were measured by semi-quantitative reverse transcription-polymerase chain reaction. RESULT: A positive correlation between TGF-beta1 and ECM were found in the present study. FN, Col IV and TGF-beta1 mRNA were inhibited by Shenluotong significantly. CONCLUSION: Shenluotong can decrease the accumulation of ECM and inhibit the expression of TGF-beta1, suggesting further that shenluotong can be used to prevent and treat various glomerular diseases and delay glomerular sclerosis.

[Effects of Semen Sojae Preparatum on lipid metabolism in ovariectomized rat].

OBJECTIVE: To investigate the antiatherosclerotic effect of Semen Sojae Preparatum (SSP) and its mechanism. METHODS: 48 rats were randomly divided into sham operation group (sham), ovariectomized group (OVX) and three doses of OVX + SSP groups. SSP was given to OVX + SSP groups 2 weeks after ovariectomy. The levels of serum lipids and lipid peroxidation were detected after 12 weeks treatment. RESULTS: SSP lowered serum oxidized-low density lipoprotein cholesterol (OX-LDL, P < 0.01), triglyceride (TG, P < 0.01) and malondialdehyde (MDA, P < 0.05 or P < 0.01), and enhanced serum high density lipoprotein cholesterol (HDL-c, P < 0.05 or P < 0.01), polipoprotein A I (apo-AI, P < 0.01) and superoxide dismutase (SOD, P < 0.01) activity. CONCLUSION: SSP had effects on regulating lipid and antioxidation. Therefore, it might inhibit atherosclerosis in ovariectomy rats.