LncRNA 1500026H17Rik knockdown ameliorates high glucose-induced mouse podocyte injuries through the miR-205-5p/EGR1 pathway.

BACKGROUND: Podocyte injuries and dysfunctions contribute to the development of diabetic nephropathy (DN). This study aimed to investigate the role and novel mechanism of lncRNA 1500026H17Rik in high glucose (HG)-treated podocytes. METHODS: DN mice were induced by streptozotocin, and DN in vitro models were constructed in mouse podocytes treated with HG. The expression of fibrosis-related proteins and early growth response protein 1 (EGR1) was detected by western blot. The expression of 1500026H17Rik, miR-205-5p and EGR1 mRNA was measured by quantitative real-time polymerase chain reaction (qRT-PCR). Cell apoptosis was monitored by flow cytometry assay. Oxidative stress was assessed according to the levels of superoxide dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species (ROS). Inflammatory response was assessed according to the releases of interleukin-6 (IL-6), interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α). The target relationship between miR-205-5p and 1500026H17Rik or EGR1 was validated by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and RNA pull-down assay. RESULTS: 1500026H17Rik was upregulated in DN mice and HG-induced podocytes. 1500026H17Rik knockdown alleviated podocyte apoptosis, fibrosis, oxidative stress and inflammation induced by HG. MiR-205-5p was a target of 1500026H17Rik, and EGR1 was a downstream target of miR-205-5p. Rescue experiments presented that miR-205-5p inhibition reversed the effects of 1500026H17Rik knockdown. Moreover, miR-205-5p restoration also ameliorated HG-induced cell injuries, which were overturned by EGR1 overexpression. In addition, EGR1 overexpression recovered podocyte apoptosis, fibrosis, oxidative stress and inflammation weakened by 1500026H17Rik knockdown. CONCLUSION: 1500026H17Rik knockdown alleviated HG-induced podocyte injuries, including apoptosis, fibrosis, oxidative stress and inflammation, by governing the miR-205-5p/EGR1 pathway, thus involving in DN development.

Letrozole Rat Model Mimics Human Polycystic Ovarian Syndrome and Changes in Insulin Signal Pathways.

BACKGROUND The aim of this study was to explore whether letrozole and high-fat diets (HFD) can induce obese insulin-resistant polycystic ovary syndrome (PCOS) with all reproductive and metabolic phenotypes in a rat model. MATERIAL AND METHODS Twenty-four 3-week-old female Sprague-Dawley rats were randomized into 4 groups: control, Letrozole, HFD, and Letrozol+HFD. The PCOS model was induced by 12 weeks of Letrozole treatment (1 mg/kg p.o. dissolved in 0.5% CMC solutions once daily) and HFD. Ovarian morphology, estrous cyclicity, hormonal status, body weight, glucose and insulin tolerance, lipid profile, and insulin signaling pathway were investigated. RESULTS The rat model manifests anovulatory cycles and PCO morphology, increased body weight, elevated testosterone levels, abnormal glucose and lipid metabolism, and insulin resistance. The rat model also expresses significantly decreased phosphorylation of 6 essential signaling proteins - INSR, IRS, PI3K, AKT, ERK1, ERK2 - in the PI3K/AKT and MAPK/ERK pathways in the classic insulin-sensitive tissues (e.g., quadriceps femoris muscle, omentum majus, and liver), as well as non-classic target ovary tissues. Disrupted insulin signaling contributes to the decrease in insulin sensitivity and compensatory hyperinsulinemia in PCOS rats. CONCLUSIONS Continuous administration of letrozole and high-fat diets can induce PCOS, metabolic phenotypes, and disrupted activation of the insulin signaling pathway.

[Effect of combined treatment with acupuncture, moxibustion and medication on endometrial receptivity and expression of serum HOXA10 in polycystic ovary syndrome of kidney deficiency and blood stagnation].

OBJECTIVE: To compare the clinical effect of the combined treatment of acupuncture, moxibustion, Chinese herbal medicine and western medication and simple western medication on polycystic ovary syndrome (PCOS) of kidney deficiency and blood stagnation pattern and explore the effect on endometrial receptivity and the expression of serum homeobox gene A10 (HOXA10). METHODS: A total of 60 patients with PCOS of kidney deficiency and blood stagnation pattern were randomized into a combined treatment group and a western medication group, 30 cases in each one. In the western medication group, on the fifth day of menstruation, clomiphene citrate tablets were taken orally, 50 mg each time, once daily, consecutively for 5 days. On the day when the follicle diameter was ≥ 18 mm, chorionic gonadotrophin for muscular injection, a dose of 10 000 U was given. Before sleep, the aspirin enteric-coated tablets were taken orally, 50 mg (except during menstruation). In the combined treatment group, on the base of the treatment as the western medication group, acupuncture and moxibustion were adopted and the Chinese herbal for tonifying the kidney and activating blood circulation was taken orally. The acupoints were Guanyuan (CV 4), Qihai (CV 6), Zusanli (ST 36), Sanyinjiao (SP 6), Zigong (EX-CA 1), etc. Acupuncture was remained for 30 min each time, once every two days and discontinued during menstruation. Chinese herbal was given from the 3rd day of menstruation till the onset of the next menstruation, one dose each day. After consecutive treatment for 3 menstrual cycles in the two groups, the real-time polymerase chain reaction (RT-PCR) method was adopted to determine the expression of serum HOXA10 before and after treatment in the patients of the two groups. The endometrial thickness at ovulatory phase, uterine arterial flow 7 days after ovulation [including uterine arterial pulsatility index (PI), resistance index (RI), peak systolic velocity (PSV)/end diastolic velocity (EDV), meaning S/D], pregnancy rate and the score of Chinese medicine symptoms before and after treatment were compared in the patients between the two groups. RESULTS: ① After treatment, the expression of serum HOXA10 was higher than that before treatment in the patients of the two groups (P<0.01). The increase range in the combined treatment group was larger than the western medication group (P<0.01). ② After treatment, the endometrial thickness at the ovulatory phase was increased in the patients of the two groups (P<0.01). The increase range of the endometrial thickness in the combined treatment group was larger than the western medication group (P<0.01). ③ The pregnancy rate was 46.7% (14/30) in the combined treatment group, higher than 26.7% (8/30) in the western medication group (P<0.05). ④ After treatment, the bilateral uterine arterial PI, RI and S/D values, as well as TCM symptom score were all lower than those before treatment in the patients of the two groups (P<0.01). The decrease range of each index in the combined treatment group was greater than the western medication group (P<0.01, P<0.05). CONCLUSION: The combined treatment with acupuncture, moxibustion and medication effectively improves endometrial receptivity and uterine arterial flow in the patients with PCOS of kidney deficiency and blood stagnation pattern and increases pregnancy rate. The therapeutic effect is better than the simple western medication and its mechanism is probably related to the regulation of serum HOXA10 expression.

[Acupuncture and Chinese medicine of artificial cycle therapy for insulin resistance of polycystic ovary syndrome with phlegm damp type and its mechanism].

OBJECTIVE: To explore the effect difference between acupuncture combined with Chinese medicine of artificial cycle therapy on the basis of western medication and simple western medication for polycystic ovarian syndrome (PCOS) of phlegm damp type with insulin resistance, and to explore its mechanism. METHODS: Sixty patients were randomly assigned into an observation group and a control group, 30 cases in each group. Patients of the control group began to take letrozole and metformin orally on the third menstrual day. Patients of the observation group were treated with acupuncture combined with Chinese medicine of artificial cycle therapy on the basis of the treatment of the control group. The main acupoints were Guanyuan (CV 4), Qihai (CV 6) and bilateral Zigong (EX-CA 1), Luanchao (Extra), Fenglong (ST 40), Yinlingquan (SP 9), Zusanli (ST 36), Sanyinjiao (SP 6). Dahe (KI 12) was matched in the follicular phase, and warm acupuncture was applied at Guanyuan (CV 4) and Qihai (CV 6); Zhongji (CV 3) and Xuehai (SP 10) were matched in the ovulatory phase, and electroacupuncture were used at Qihai (CV 6) and Zhongji (CV 3), Xuehai (SP 10) and Yinlingquan (SP 9), 2 Hz and continuous wave; The acupoints in the luteal phase were the above main acupoints, at the same time, moxibustion was used at Guanyuan (CV 4) and Qihai (CV 6), the treatment was given once every other day, 30 min a time. There was no treatment in the menstrual phase. Chinese medicine was applied for tonifying kidney and nourishing blood, strengthening spleen and eliminating phlegm. The patients of the two groups were treated for continuous three menstrual cycles. The indexes before and after treatment were detected, including serum microRNA-29 (miR-29) expression, TCM symptom score, insulin (INS), blood-sugar content and homeostasis model assessment-insulin resistance (HOMA-IR) index, the endocrine hormone values of follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), LH/FSH ratio, prolactin (PRL). The pregnancy rates of the two groups were compared. RESULTS: After treatment the expressions of miR-29 and TCM symptom scores of the two groups were lower than those before treatment (P<0.01, P<0.05), with better results in the observation group (P<0.01, P<0.05). The levels of LH LH/FSH and T of the observation group were lower than those before treatment (P<0.01, P<0.05), and the T value in the control group decreased after treatment (P<0.05). The changes of LH and LH/FSH in the observation group were better than those in the control group (both P<0.05). After treatment, The INS, blood-sugar content and HOMA-IR were lower than those before treatment in the two groups (P<0.05, P<0.01), with better results for INS and HOMA-IR improvements in the observation group (both P<0.05). The pregnancy rate in the observation group was 56.7% (17/30), which was significantly higher than 30.0% (9/30) in the control group (P<0.05). CONCLUSION: Acupuncture combined with Chinese medicine of artificial cycle therapy can improve the endocrine levels and insulin resistance of PCOS with phlegm damp type. At the same time, the pregnancy rate is improved. By the view of molecular biology, there may be a pathway trough changing serum miR-29 expression.