The Mechanism of TNF-α-Mediated Accumulation of Phosphorylated Tau Protein and Its Modulation by Propofol in Primary Mouse Hippocampal Neurons: Role of Mitophagy, NLRP3, and p62/Keap1/Nrf2 Pathway.

Background: Neuroinflammation-induced phosphorylated Tau (p-Tau) deposition in central nervous system contributes to neurodegenerative disorders. Propofol possesses neuroprotective properties. We investigated its impacts on tumor necrosis factor-α (TNF-α)-mediated p-Tau deposition in neurons. Methods: Mouse hippocampal neurons were exposed to propofol followed by TNF-α. Cell viability, p-Tau, mitophagy, reactive oxygen species (ROS), NOD-like receptor protein 3 (NLRP3), antioxidant enzymes, and p62/Keap1/Nrf2 pathway were investigated. Results: TNF-α promoted p-Tau accumulation in a concentration- and time-dependent manner. TNF-α (20 ng/mL, 4 h) inhibited mitophagy while increased ROS accumulation and NLRP3 activation. It also induced glycogen synthase kinase-3ß (GSK3ß) while inhibited protein phosphatase 2A (PP2A) phosphorylation. All these effects were attenuated by 25 µM propofol. In addition, TNF-α-induced p-Tau accumulation was attenuated by ROS scavenger, NLRP3 inhibitor, GSK3ß inhibitor, or PP2A activator. Besides, compared with control neurons, 100 µM propofol decreased p-Tau accumulation. It also decreased ROS and NLRP3 activation, modulated GSK3ß/PP2A phosphorylation, leaving mitophagy unchanged. Further, 100 µM propofol induced p62 expression, reduced Keap1 expression, triggered the nuclear translocation of Nrf2, and upregulated superoxide dismutase (SOD) and heme oxygenase-1 (HO-1) expression, which was abolished by p62 knockdown, Keap1 overexpression, or Nrf2 inhibitor. Consistently, the inhibitory effect of 100 µM propofol on ROS and p-Tau accumulation was mitigated by p62 knockdown, Keap1 overexpression, or Nrf2 inhibitor. Conclusions: In hippocampal neurons, TNF-α inhibited mitophagy, caused oxidative stress and NLRP3 activation, leading to GSK3ß/PP2A-dependent Tau phosphorylation. Propofol may reduce p-Tau accumulation by reversing mitophagy and oxidative stress-related events. Besides, propofol may reduce p-Tau accumulation by modulating SOD and HO-1 expression through p62/Keap1/Nrf2 pathway.

[Effect of electroacupuncture on tissue elasticity and expression of muscular MyoD and myogenin in rats with acute gastrocnemius contusion].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Ashi" acupoint and "Kunlun" (BL60) on elastic modulus, histopathological changes and expression of myogenic regulatory factors in gastrocnemius(GM) contusion rats, so as to explore the therapeutic effect of local acupoint selection and acupoint selection along channel. METHODS: Male SD rats were randomly divided into blank control (n＝5), model (n＝15), Ashi-point (n＝15) and BL60 (n＝15) groups. The acute GM contusion model was established by striking (free falling) the GM with a homemade hitter. EA (0.5 to 1.0 mA, 2 Hz/10 Hz) was applied to Ashi-point (local focus) and BL60 for 30 min 24 h after muscle injury. The elasticity maximum (Emax) of gastrocnemius muscle was measured by using an ultrasonic device. Histopathological changes were observed after H．E. stain, and the number of Myogenic differentiation(MyoD)- and Myogenin (MyoG)-positive cells was detected by using immunohistochemistry. RESULTS: After mdeling, the Emax value of GM was significantly increased from the 3rd h to 7th day in comparison with pre- injury of muscle (P<0.05), and was markedly increased on the 3rd day and obviously lower on day 7 in the Ashi-point group than in the model group (P<0.05). The numbers of MyoD- and MyoG-positive cells of GM were significantly increased on day 7 in the model group than in the blank control group (P<0.05), and both further increased in Ashi-point on day 3 and 5, and MyoG-positive cells further increased in BL60 group on day 5 and in Ashi-point group on day 7 relevant to the model group(P<0.05). The therapeutic effect of EA-Ashi-point was apparently superior to that of BL60 in up-regulating Emax on day 3 and in up-regulating the number of MyoD-positive cells on day 3 and 5 (P<0.05). H．E. stain showed disordered arrangement of muscle fibers, infiltration of inflammatory cells, increase of intercellular space, and edema on day 3 after modeling (which was milder in the Ashi-point group); and gradual fusion and thickening of new born muscle fibers with obvious connective tissue hyperplasia converged to the lesioned region on day 7 in the model group (convergence of new born muscle cells to the lesion region in both EA groups, and more complete tissues in the Ashi-point group). CONCLUSION: EA of Ashi-point and BL60 can up-regulate the expression of myogenic regulatory factors MyoD and MyoG of GM tissue in GM contusion rats, which may contribute to its function in promoting recovery of muscle elasticity. The role of EA-Ashi-point is superior to that of EA-BL60.

[Effect of moxibustion pretreatment at different time on ovarian function in rats with dimini-shed ovarian reserve].

OBJECTIVE: To observe the effect of moxibustion pretreatment at different time on serum hormone levels in diminished ovarian reserve (DOR) rats, so as to explore its protective mechanisms. METHODS: Forty female SD rats were randomly divided into control, model, moxibustion-1 (moxibustion was given 4 weeks before modeling), moxibustion-2 (moxibustion was given 2 weeks before modeling and 2 weeks from the 1st day on after modeling ) and moxibustion-3 (moxibustion was given 4 weeks from the 1st day on after modeling) groups (n=8 rats in each group). The DOR model was established by gavage of Tripterygium Glycosides (75 mg/kg) once daily for 14 days. Grain-moxibustion was applied to "Shenshu" (BL23) and "Guanyuan" (CV4) for 7 cones, 5 times a week for 4 weeks. The body weight and the ovary weight were recorded for calculating the ovarian index. The levels of serum anti-mullerian hormone (AMH), follicle stimulating hormone (FSH), estradiol (E2), androgen (T) and dehydroepiandrosterone (DHEA) were detected by ELISA. RESULTS: After modeling, ovarian index and serum AMH levels were obviously decreased (P<0.05), and the levels of serum FSH, E2, T and DHEA were significantly increased in contrast with the control group (P<0.05). Following intervention and compared with the model group, the serum FSH and DHEA levels of each moxibustion group were significantly reduced (P<0.05), the AMH levels significantly increased and E2 and T contents significantly decreased in the moxibustion-2 and moxibustion-3 groups (P<0.05). The serum FSH, E2 and T contents in moxibustion-2 group were obviously lower than those of the moxibustion-1 and moxibustion-3 groups (P<0.05). CONCLUSION: Moxibustion pre-treatment can improve ovarian reserve function in DOR rats, while the effect is different with different intervention time, and the best intervention time is pre-occurrence and early stage of DOR.

Correction: Acupuncture with reinforcing and reducing twirling manipulation inhibits hippocampal neuronal apoptosis in spontaneously hypertensive rats.

[This corrects the article on p. 770 in vol. 12, PMID: 28616034.].

Acupuncture with reinforcing and reducing twirling manipulation inhibits hippocampal neuronal apoptosis in spontaneously hypertensive rats.

To observe the effects of different acupuncture manipulations on blood pressure and target organ damage in spontaneously hypertensive rats (SHRs), this study used the reinforcing twirling method (1.5-2-mm depth; rotating needle clockwise for 360° and then counter clockwise for 360°, with the thumb moving heavily forward and gently backward, 60 times per minute for 1 minute, and retaining needle for 9 minutes), the reducing twirling method (1.5-2-mm depth; rotating needle counter clockwise for 360° and then clockwise for 360°, with the thumb moving heavily backward and gently forward, 60 times per minute for 1 minute, and retaining needle for 9 minutes), and the needle retaining method (1.5-2-mm depth and retaining the needle for 10 minutes). Bilateral Taichong (LR3) was treated by acupuncture using different manipulations and manual stimulation. Reinforcing twirling, reducing twirling, and needle retaining resulted in a decreased number of apoptotic cells, reduced Bax mRNA and protein expression, and an increased Bcl-2/Bax ratio in the hippocampus compared with the SHR group. Among these groups, the Bcl-2/Bax protein ratio was highest in the reducing twirling group, and the Bcl-2/Bax mRNA ratio was highest in the needle retaining group. These results suggest that reinforcing twirling, reducing twirling, and needle retaining methods all improve blood pressure and prevent target organ damage by increasing the hippocampal Bcl-2/Bax ratio and inhibiting cell apoptosis in the hippocampus in SHR.

Effects of acupuncture applied to sanyinjiao with different stimuli on uterine contraction and microcirculation in rats with dysmenorrhea of cold coagulation syndrome.

In this study, we try to evaluate the effects of acupuncture stimulation with different amounts at Sanyinjiao (SP6) on uterine contraction and uterus microcirculation in rats with dysmenorrhea of cold coagulation syndrome and to explore whether there is direct relativity between "De qi" and needle stimulus intensity. Diestrus female rats were randomly divided into 4 groups, including saline control group, model control group, "A" stimulus group (with strong stimulus), and "B" stimulus group (with weak stimulus). We found that dysmenorrhea rats of the cold coagulation syndrome present a high intensity in uterine tension and high contraction of microvascular diameter. Acupuncture applied with two different stimuli could relieve the symptoms, but, compared with "B" stimulus, "A" stimulus leads to better outcomes on reducing uterine contraction and increasing diameter of uterine microvascular; moreover, hand manipulation during needling mediates the curative effect on the microvascular diameter. Our finding indicates that using thick needles and deep insertion with hand manipulation are more effective and achieve desired level of "De qi" in dysmenorrhea rats.

Effects of preventive acupuncture and moxibustion on fat accumulation, blood lipid, and uterus e 2 of menopause rats.

Objective. To observe the effect of preventive acupuncture and moxibustion on blood lipid of menopause rats. Methods. Seventy 10-month-old SD rats with estrous cycle disorders were divided into three control groups and four treatment groups (n = 10/group) and another ten 3.5-month-old female SD rats were chosen as young control group. Preventive acupuncture and moxibustion were applied at Guanyuan (CV 4). Body weight growth rate has been recorded. Plasma total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL), and high density lipoprotein (HDL) levels and uterus E 2 level were measured. Results. Compared to young control group, plasma TC and LDL increased and uterus E 2 reduced significantly in 12-month-old control group. Compared to 12-month-old control group, plasma TC and LDL level and body weight growth rate decreased while HDL level increased remarkably in preventive acupuncture 12-month-old group. Compared to 14-month-old control group, plasma TC level and body weight growth rate decreased remarkably in preventive moxibustion 14-month-old group. Conclusions. Preventive acupuncture and moxibustion can significantly decrease the plasma TG and LDL, increase the plasma HDL, and prevent fat accumulation. Our finding suggests that preventive acupuncture and moxibustion have beneficial effects on blood lipid. Different treatment effects were found between preventive acupuncture and preventive moxibustion.

The Effect of Acupuncture to SP6 on Skin Temperature Changes of SP6 and SP10: An Observation of "Deqi".

Background. Deqi sensation is a complex but an important component for acupuncture effect. In this study, we tried to observe the relationship between Deqi and skin temperature changes and whether there was some relativity between Deqi and needle stimulations on cold congealing and dysmenorrhea rat model. Thirty-two female Sprague Dawley (SD) rats were randomly divided into four groups (Saline Control Group, Model Group, Group A with strong stimulation, and Group B with small stimulation). Group A and Group B were performed with different stimulations. We found that, compared with saline control group, model group, and Group B, Group A showed that the skin temperature changes on right acupoint SP6 and SP10 increased significantly at 5 min-10 min interval. The skin temperature changes on left SP6 decreased at instant-5 min interval. The skin temperature changes on right SP10 decreased significantly at instant-5 min interval and 10 min-20 min interval. Thermogenic action along Spleen Meridian of Foot Greater Yin was manifested as simultaneous skin temperature increase on right SP6 and SP10 at 5 min-10 min interval after needling SP6, which was helpful to illustrate the relationship between the characteristic of Deqi and needle stimulations.

Acupuncture De-qi: From Characterization to Underlying Mechanism.

De-qi refers to the participant's subjective sensations and objective body responses as well as the acupuncturist's perceptions while the acupuncturist needles certain acupoints in the participant's body. In recent years, De-qi is getting increasing attention of the researchers and many efforts have been made to understand its mechanism. By the broad literature survey, this paper explores the subjective De-qi sensation of the patients, its influencing factors, and the resulting physiological responses. The purpose of this paper is expected to find out a possible mechanism of De-qi and to provide certain scientific evidence for acupuncture fundamental research and clinical practice.