Electroacupuncture Alleviates Bladder Overactivity via Inhabiting Bladder P2X3 Receptor.

Electroacupuncture (EA) has been widely applied for overactive bladder, but the mechanism of its action remains to be clarified. This study was aimed to investigate EA regulating the effect of purinergic signaling in the OAB of rats. Electroacupuncture (continuous wave, 30 Hz, 1 mA) was applied to stimulate the Ciliao point (BL32) and the Huiyang point (BL35) of rats. Results showed that when the P2X3 receptor in bladder peripheral level and the spinal cord central level was involved in the bladder micturition reflex of the afferent signaling, intravenous administration P2X3 antagonist AF-353 can significantly inhibit urination in naive rats and OAB of rats and increase bladder volume and micturition pressure. EA stimulation alleviated bladder overactivity significantly and after the P2X3 receptor was blocked, the EA effect was weakened. EA stimulation can effectively reduce the P2X3 mRNA and protein expression in OAB of rats, spinal cord (L6-S1), and DRG (L6-S1) and can significantly reduce the number of positive P2X3 cells in OAB of rats, spinal cord (L6-S1), and DRG (L6-S1). These findings suggest that EA stimulation could alleviate bladder overactivity, and the function is closely related to the inhabited P2X3 receptor in the bladder.

[Effect of electroacupuncture at different acupoints on follicle development and related factors in serum and ovary tissues of PCOS rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at different acupoints on follicle development, expression of gonadotropins and their receptors and anti-Müllerian hormone(AMH), inhibin B(INHB) in polycystic ovarian syndrome (PCOS) rats, so as to explore its mechanisms underlying improvement of PCOS. METHODS: Sixty female SD rats were randomized into six groups: control, model, Zusanli(ST36), Sanyinjiao(SP6), Guanyuan(CV4) and combination (ST36+SP6+CV4, n=10 rats/group). The PCOS model was established by gavage of Letrozole solution (1.0 mg/kg) once daily for 21 consecutive days. Rats of the control group were given 1% Carboxymethyl Cellulose (CMC, 1 mg/kg). EA (2 Hz) was applied to ST36, SP6, or/and CV4 for 20 min, once daily for 14 consecutive days. The number of follicles was counted, and the ovarian structure and follicular development were observed under light microscope after H.E. stain, and the contents of serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), AMH, and INHB were assayed by enzyme-linked immunosorbent assay, and the ratio of LH/FSH was calcula-ted. The immunoactivity of LH receptor (LHR) and FSH receptor (FSHR) proteins was detected by immunohistochemistry. RESULTS: After modeling, the number of follicles at the growth stage, contents of serum LH, AMH and INHB, and ratio of LH/FSH were significantly increased, and serum FSH level and FSHR, LHR immunoactivity were remarkably decreased in the model group relevant to the control group (P<0.05, P<0.01). Following EA intervention, the number of follicles at the growth stage in the SP6, CV4 and combination groups, LH/FSH ratio and serum AMH levels in the 4 EA groups, INHB contents in the ST36, CV4 and combination groups were significantly reduced (P<0.05, P<0.01), while serum FSH contents in the 4 EA groups, FSHR immunoactivity at the early stage in the ST36 group and LHR immunoactivity at both early and late stages in the ST36 and CV4 groups were considerably increased in comparison with those of the model group (P<0.01, P<0.05). The therapeutic effect of SP6 was significantly superior to that of CV4 in down-regulating serum LH level (P<0.01), but significantly inferior to that of CV4 in up-regulating serum FSH content and in down-regulating LH/FSH ratio and serum AMH and INHB levels (P<0.05, P<0.01). The effect of CV4 was comparable to that of ST36 in up-regulating serum FSH, and in down-regulating serum LH/FSH ratio, AMH and INHB levels (P>0.05). CONCLUSION: EA of CV4, SP6, ST36 and ST36+CV4+SP6 can reduce the number of follicles at the growth stage and regulate the expression levels of gonadotropins in PCOS rats. The effects of EA of CV4 and ST36 are evidently better than those of EA of SP6 in up-regulating serum FSH content and in down-regulating LH/FSH ratio, and serum AMH and INHB levels, and EA of SP6 is evidently superior to EA of CV4 down-regulating LH level, but without synergistic effect among the 3 acupoints.

[Effect of Electroacupuncture at Different Acupoints on Steroid Hormones and Ovarian Androgen Receptor Expression in Polycystic Ovary Syndrome Rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at different acupoints on hyperandrogenism and ovarian androgen receptor (AR) expression in rats with polycystic ovary syndrome (PCOS), so as to explore its mechanisms underlying improvement of PCOS. METHODS: Sixty female SD rats were randomly divided into control, model, Zusanli (ST 36), Sanyinjiao (SP 6), Guanyuan (CV 4) and comprehensive groups (10 rats/group). The PCOS model was established by giving (gavage) the animals with Letrozole solution (1.0 mg/kg), once daily for 21 consecutive days. EA stimulation was applied to ST 36, SP 6, or CV 4 respectively, once daily for 14 days. The comprehensive group was treated by EA treatment of the abovementioned three acupoints in the meantime. Before and after the treatment, the body mass and the ovary weight were measured. The morphological structure of the ovary was observed under microscope after H．E. staining. The levels of serum hormone and sex hormone binding globulin (SHBG) were assayed by ELISA. Free androgen index (FAI) was calculated and the expression of AR in the ovary was detected by immunohistochemistry. RESULTS: After modeling, the body mass, ovary weight, serum testosterone (T) and FAI levels and the expression of AR protein in late ovarian follicles were significantly increased (P<0.01, P<0.05), and serum estradiol (E2) and SHBG contents were significantly decreased relevant to the control group (P<0.01). Following EA intervention and compared with the model group, the body weight of each EA group was significantly reduced (P<0.05), the ovarian pathological changes were improved, and serum T and FAI levels were significantly decreased (P<0.01), and serum E2 and SHBG contents significantly increased in the ST 36, CV 4 and comprehensive groups (P<0.01, P<0.05). The ovary weight of only CV 4 group was significantly lower than that of the model group (P<0.01). The expression of AR protein in late follicles of both CV 4 and SP 6 groups was markedly decreased (P<0.05, P<0.01). CONCLUSION: EA at ST 36, SP 6, and CV 4 all can improve the hyperandrogenism and ovarian polycystic morphological changes of PCOS rats, while different acupoints have different functional characteristics.

[Deltamethrin resistance, metabolic detoxification enzyme and kdr mutation in Anopheles sinensis in region along Huaihe River in Anhui Province].

OBJECTIVE: To understand the deltamethrin resistance level, GTS and P450 metabolic detoxification enzyme activities, and mutations in the para-type sodium channel gene that confers knockdown resistance (kdr) in Anopheles sinensis mosquitoes from malaria endemic areas along the Huaihe River in Anhui Province, China. METHODS: An. sinensis adult mosquitoes were collected in Lilou, Mohekou and Tuohu townships of Bengbu City, Anhui Province from August to September, 2011. The insecticide resistance bioassays were performed on adult mosquitoes by using the standard WHO susceptibility test with diagnostic concentrations of deltamethrin 0.05%, and the mosquito resistance status was classified based on the WHO resistance classification criteria. The metabolic detoxification enzyme activities were measured in randomly selected mosquitoes, and the IIS6 region of the para-type sodium channel gene was amplified by PCR and sequenced to detect mutations at the codon 1014. RESULTS: The knockdown rates within 60 min exposure to deltamethrin test paper were 4.1%, 7.0% and 8.2%, and the mortality rates were 8.2%, 12.0% and 12.8% for mosquitoes collected from Lilou, Mohekou and Tuohu townships, respectively. These three populations were classified as highly resistant populations based on the WHO resistance classification criteria. The GST and P450 enzyme activities of the three populations were significantly higher than those of the susceptible laboratory population (P < 0.001). L1014C and L1014F mutations were detected, and the wild type homozygote kdr genotype was not found. These three populations exhibited a small but insignificant difference in kdr allele frequencies. No mutation was found in the laboratory susceptible mosquitos. CONCLUSION: The An. sinensis mosquito populations from the regions along the Huaihe River in Anhui Province are strongly resistant to pyrethroid insecticides, and exhibit significantly higher metabolic detoxification enzyme activities than the laboratory susceptible population. The high frequency of kdr mutation is identified in the mosquito populations from the region along the Huaihe River in Anhui Province, China.