[Effect of wheat-grain moxibustion on the expression of Beclin-1/GRP78 in spinal dorsal horn in rats with cervical spondylotic radiculopathy].

OBJECTIVE: To observe the effect of wheat-grain moxibustion at "Dazhui" (GV 14) on the expressions of Beclin-1 and GRP78 in spinal dorsal horn in rats with cervical spondylotic radiculopathy (CSR), and to explore the possible analgesic mechanism of wheat-grain moxibustion for CSR. METHODS: A total of 48 SD rats were randomly divided into a sham operation group, a model group, a wheat-grain moxibustion group and a wheat-grain moxibustion+3-MA group, 12 rats in each group. The CSR model was prepared by spinal cord insertion method. Three days after modeling, the rats in the model group were intraperitoneally injected with 1 mL of 0.9% sodium chloride solution; the rats in the wheat-grain moxibustion group were treated with wheat-grain moxibustion at "Dazhui" (GV 14, 6 cones per time) on the basis of the model group; the rats in the wheat-grain moxibustion+3-MA group were intraperitoneally injected with 3-MA solution and wheat-grain moxibustion at "Dazhui" (GV 14, 6 cones per time). The three groups were intervened for 7 days, once a day. The gait score and mechanical pain threshold were observed before treatment and 7 days into treatment; after the treatment, the expressions of mRNA and protein of Beclin-1 in spinal dorsal horn were detected by real-time fluorescence quantitative PCR and immunohistochemistry; the expression of GRP78 protein in spinal dorsal horn was detected by Western blot method; the autophagosomes and ultrastructure in spinal dorsal horn neurons were observed by electron microscope. RESULTS: After the treatment, compared with the sham operation group, in the model group, the gait score was increased and the mechanical pain threshold was decreased (P<0.01), and the expression of GRP78 protein in spinal dorsal horn was increased (P<0.01). Compared with the model group and the wheat-grain moxibustion+3-MA group, in the wheat-grain moxibustion group, the gait score was decreased and mechanical pain threshold was increased (P<0.01), and the expression of GRP78 protein in spinal dorsal horn was decreased, and the expressions of mRNA and protein of Beclin-1 were increased (P<0.01). Under electron microscope, the ultrastructure of spinal dorsal horn neurons in the wheat-grain moxibustion group was not significantly damaged, and its structure was basically close to normal, and the number of autophagosomes was more than the other three groups. CONCLUSION: Wheat-grain moxibustion at "Dazhui" (GV 14) has analgesic effect on CSR rats. The mechanism may be related to moderately up-regulate the expression of Beclin-1, enhance autophagy and reduce endoplasmic reticulum stress.

[Effect of mild moxibustion on pain and expression of LC3 and Bax in spinal cord in cervical spondylotic radiculopathy rats].

OBJECTIVE: To observe the effect of mild moxibustion (Moxi) at "Dazhui" (GV14) on neuropathic pain, expression of autophagy and apoptosis factor LC3 and Bax proteins and mRNAs in the spinal cord tissue in rats with cervical spondylotic radiculopathy (CSR), so as to explore its underlying mechanism underlying relief of CSR-induced pain. METHODS: Forty rats (half male half female) were randomly divided into blank control, model, Moxi, Moxi+autophagy inhibitor 3-methyladenine (3-MA, Moxi+3-MA) groups, with 10 rats in each group. The CSR model was established by loose ligature of the local cervical nerve roots. Three days after modeling, mild Moxi was applied to GV14 for 10 min, once daily for 7 days. Rats of the Moxi+3-MA group received intraperitoneal injection of 3-MA(1 mL, 15 mg/kg+ saline) before Moxi, once daily for 7 consecutive days. Rats of the model and Moxi groups were also given normal saline (i.p., 1 mL), once daily for 7 days. The gait behavior score (1-3 points) was scaled according to the rats' pain reaction and foot paw contracture produced walking disorder and the mechanical pain threshold (MPT) was detected before and after the treatment. The expression of spinal cord LC3 and Bax proteins and mRNAs were detected by immunohistochemistry and quantitative RT-PCR, respectively. RESULTS: Compared with the blank control group, the gait disorder score, and percentage of Bax positive cells and expression of Bax mRNA were significantly increased (P<0.01, P<0.05), and MPT was markedly decreased in the model group (P<0.01). After the treatment, the gait disorder score, percentage of Bax positive cells and Bax mRNA expression were significantly down-regulated (P<0.01, P<0.05), while the MPT and percentage of LC3 positive cells and LC3 mRNA expression were considerably increased (P<0.01, P<0.05) in both Moxi and Moxi+3-MA groups. The therapeutic effects of mild Moxi were remarkably superior to those of Moxi+3-MA in downregulating gait disorder score, Bax positive cell percentage and Bax mRNA expression, and in up-regulating MPT, LC3 positive cell percentage and LC3 mRNA expression (P<0.05), suggesting a reduction of the function of mild Moxi after administration of 3-MA. CONCLUSION: Mild Moxi at GV14 can relieve neuropathic pain in CSR rats, which may be related to its functions in up-regulating LC3 autophagy, thereby inhibiting the expression of Bax pro-apoptotic protein in spinal cord to reduce apoptosis and to repair nerve injury.

Direct moxibustion exerts an analgesic effect on cervical spondylotic radiculopathy by increasing autophagy via the Act A/Smads signaling pathway.

BACKGROUND: Direct moxibustion (DM) is reported to be useful for cervical spondylotic radiculopathy (CSR), but the analgesic mechanism remains unknown. Autophagy plays a protective role in neuronal apoptosis, Act A/Smads signaling pathway has been confirmed to be associated with the activation of autophagy. The study aimed to explore the effect of DM on autophagy in rats with CSR and the involvement of Act A/Smads signaling pathway. METHODS: Rats were randomly divided into Sham, CSR, CSR + DM, CSR + DM + 3-MA (PI3K inhibitor), and CSR + DM + SB (Act A inhibitor) group. Three days after establishment of CSR model with a fish line inserted under the axilla of the nerve roots, DM at Dazhui (GV14) was performed six times once for seven consecutive days. Western blot and immunofluorescence staining were used to observe the expression of the neuronal autophagy molecule LC3II/I, Atg7, and Act A/Smads signaling molecule Act A, p-Smad2, and p-Smad3. Bcl-2/Bax mRNA expression was measured by real time PCR. RESULTS: DM improved the pain threshold and motor function of CSR rats and promoted the expression of Act A, p-Smad2, p-Smad3, LC3II/I, and Atg7 in the entrapped-nerve root spinal dorsal horn. DM reduced the expression of Bax mRNA and decreased the number of apoptotic neurons. 3-MA and Act A inhibitor SB suppressed the expression of above-mentioned proteins and reduced the protective effect of DM on apoptotic neurons. CONCLUSION: DM exerts analgesic effects by regulating the autophagy to reduce cell apoptosis and repair nerve injury, and this feature may be related to the Act A/Smads signaling pathway.

[Observation on clinical effect of acupuncture combined with wheat-grain moxibustion for mild to moderate postpartum depression].

OBJECTIVE: To compare the clinical effect of acupuncture combined with wheat-grain moxibustion and oral sertraline hydrochloride dispersible tablets in the treatment of mild to moderate postpartum depression. METHODS: Sixty patients with mild to moderate postpartum depression were randomly divided into an observation group and a control group, 30 cases in each group. Both groups were treated with psychotherapy. The control group was treated with oral sertraline hydrochloride dispersible tablets, 50 mg each time, once a day; the observation group was treated with acupuncture at Qihai (CV 6), Zusanli (ST 36), Xuehai (SP 10), Hegu (LI 4), Sanyinjiao (SP 6), Taixi (KI 3), etc. combined with wheat-grain moxibustion at Xinshu (BL 15), Pishu (BL 20), Ganshu (BL 18) and Shenshu (BL 23), once every other day, 3 times a week. Both groups were treated for 4 weeks as a course, with 2 consecutive courses of treatment. Before and after treatment and follow-up of 3 months after the end of treatment, the Hamilton depression scale (HAMD), Edinburgh postnatal depression scale (EPDS) and World Health Organization quality of life-BREF (WHOQOL-BREF) score of the two groups were compared, and the clinical effect was assessed. RESULTS: After treatment and during follow-up, the HAMD and EPDS scores of the two groups were lower than before treatment (P<0.05), and the WHOQOL-BREF scores of the two groups were higher than before treatment (P<0.05). In the control group, the scores of HAMD and EPDS during follow-up were higher than after treatment (P<0.05), and the score of WHOQOL-BREF during follow-up was lower than after treatment (P<0.05). After treatment and during follow-up, the HAMD and EPDS scores of the observation group were lower than those of the control group (P<0.05), and the WHOQOL-BREF score of the observation group was higher than that of the control group (P<0.05). The total effective rate of the observation group was 93.3% (28/30), which was higher than 86.7% (26/30) of the control group (P<0.05). CONCLUSION: Acupuncture combined with wheat-grain moxibustion can improve the depressive symptoms of patients with mild to moderate postpartum depression and improve their quality of life, and the clinical effect is more lasting and stable than oral sertraline hydrochloride dispersible tablets.

[Effect of auricular acupuncture on reduction of sertraline hydrochloride in patients with depression: a randomized controlled trial].

OBJECTIVE: To observe the effect of auricular acupuncture on reduction rate of sertraline hydrochloride, and to explore the long-term efficacy of auricular acupuncture in patients with depression. METHODS: Seventy-two patients with depression were randomly divided into an observation group (36 cases, 1 case dropped off) and a control group (36 cases, 2 cases dropped off). The patients in the control group were treated with conventional dosage reduction method, that is, the dosage of sertraline hydrochloride was reduced by 25% every week until the dosage was stopped completely on the premise of no aggravation of depressive symptoms. Based on the conventional dosage reduction method used in the control group, the patients in the observation group were treated with auricular acupuncture at Xin (CO15), Gan (CO12), Shenmen (TF4), Pizhixia (AT4) and Neifenmi (CO18), once every 3 days for 4 weeks. The reduction of dosage was observed before dosage reduction, after 1, 2, 3, 4 weeks of dosage reduction and follow-up 6 months after the end of treatment; the rate of dosage reduction was observed after 4 weeks of dosage reduction and during follow-up; the Hamilton depression scale (HAMD) was used to evaluate the depression severity before dosage reduction, after 4 weeks of dosage reduction and during follow-up; the incidence rate of withdrawal syndrome and clinical efficacy were compared between the two groups. RESULTS: The dosage of sertraline hydrochloride in the observation group was less than that in the control group after 2, 3, 4 weeks of dosage reduction and during follow-up (P<0.05). The dosage reduction rates were 80.6% and 82.2% after 4 weeks of dosage reduction and during follow-up in the observation group, which were higher than 65.8% and 62.2% in the control group (P<0.05). After 4 weeks of dosage reduction, the HAMD scores in the two groups were higher than those before dosage reduction (P<0.05), and the HAMD score in the observation group was lower than that in the control group (P<0.05). During follow-up, HAMD score in the observation group was lower than that in the control group and that after 4 weeks of dosage reduction (P<0.05), while HAMD score in the control group was higher than that before dosage reduction and that after 4 weeks of treatment (P<0.05). The incidence rate of withdrawal syndrome in the observation group was 11.4% (4/35), which was lower than 47.1% (16/34) in the control group (P<0.05). After 4 weeks of dosage reduction and during follow-up, the total effective rate was 97.1% (34/35) in the observation group, which was higher than 79.4% (27/34) in the control group (P<0.05). CONCLUSION: Auricular acupuncture could effectively reduce the dosage of sertraline hydrochloride, improve the dosage reduction rate, reduce the incidence of withdrawal syndrome and reduce the risk of long-term recurrence in patients with depression.

[Clinical observation of acupuncture combined with moxibustion on college students with depression].

OBJECTIVE: To observe the clinical efficacy of acupuncture combined with moxibustion with seed-sized moxa cone in the treatment of depression in college students, and explore its effect on serum 5-hydroxytryptamine (5-HT). METHODS: Sixty undergraduates with depression were divided into acupuncture-moxibustion group and medication group according to the random number table, with 30 patients in each group. The patients in acupuncture-moxibustion group received acupuncture and moxibustion with seed-sized moxa cone at acupoints on Shaoyang meridian according to the method of "rotating the pivot and regulating the qi", one time every other day. The patients in medication group received oral administration of fluoxetine hydrochloride capsule 20 mg once a day, and both groups were treated continuously for 8 weeks. The scores of Hamilton depression (HAMD-17) scale, self-rating depression scale (SDS) and serum 5-HT content before and after treatment were compared between the two groups. RESULTS: Compared with before treatment, the HAMD-17 and SDS scores of the acupuncture-moxibustion group began to decrease after 2 weeks of treatment (P<0.05); while the HAMD-17 and SDS scores of the the medication group began to decrease after 4 weeks of treatment (P<0.05). After 2, 4 and 8 weeks of treatment, the HAMD-17 and SDS scores of the acupuncture-moxibustion group decreased more significantly than the medication group (P<0.05). The 5-HT contents of the two groups were significantly higher than those before treatment (P<0.05)，and there was no significant difference in serum 5-HT content between the two groups (P>0.05). The adverse reaction score of the acupuncture-moxibustion group was lower than that of the medication group (P<0.05). The total effective rate of the acupuncture-moxibustion group was 92.86%（26/28）, better than the medication group 81.48% (22/27,P<0.05). CONCLUSION: Acupuncture combined with seed-sized moxa cone moxibustion is more effective than oral administration of fluoxetine hydrochloride in treating college students' depression, and acupuncture combined with moxibustion has a faster onset and fewer adverse reactions in the treatment of college students' depression.

[Therapeutic effect of acupuncture combined with wheat-grain moxibustion on perimenopausal depression with kidney deficiency and liver depression].

OBJECTIVE: To compare the therapeutic effect of acupuncture combined with wheat-grain moxibustion and oral fluoxetine hydrochloride capsule on perimenopausal depression with kidney deficiency and liver depression. METHODS: A total of 60 patients of perimenopausal depression with kidney deficiency and liver depression were randomized into an observation group and a control group, 30 cases in each one. Acupuncture combined with wheat-grain moxibustion were adopted in the observation group. Acupuncture was applied at Baihui (GV 20), Yintang (GV 29), Fengchi (GB 20), etc. for 30 min. Wheat-grain moxibustion was applied at Ganshu (BL 18), Shenshu (BL 23), Mingmen (GV 4) and Yongquan (KI 1), 3 moxa-cones for each acupoint. The treatment in the observation group was given once every 2 days, 3 times a week. Fluoxetine hydrochloride capsule was given orally in the control group, 20 mg each time, once a day. Treatment for 8 weeks was required in the both groups. The scores of Hamilton depression scale (HAMD) and modified Kupperman scale were observed in the both groups before and after treatment, and at 1, 3, 6-month follow-up. The therapeutic efficacy was evaluated after treatment. RESULTS: Compared before treatment, the scores of HAMD and modified Kupperman scale after treatment and at each time point of follow-up were decreased in the both groups (P<0.01), and the HAMD scores in the observation group were lower than the control group (P<0.01). The total effective rate was 93.3% (28/30) in the observation group, which was superior to 80.0% (24/30) in the control group (P<0.01). CONCLUSION: Acupuncture combined with wheat-grain moxibustion can effectively treat perimenopausal depression with kidney deficiency and liver depression, and have more stable and sustained therapeutic effect compared with oral fluoxetine hydrochloride capsule.

[Effects of mild moxibustion on Beclin-1/Bcl-2 expression in spinal cord of rats with cervical spondylotic radiculopathy].

OBJECTIVE: To observe the effects of mild moxibustion on the expression of autophagy and apoptosis factors Beclin-1, Bcl-2 mRNA and protein in spinal cord (including nerve root tissues) of cervical spondylotic radiculopathy (CSR) rats, so as to explore the analgesic mechanism of mild moxibustion at "Dazhui" (GV14) on CSR. METHODS: SD rats were randomly divided into blank group, model group, mild moxibustion group and mild moxibustion+3-methyladenine(3-MA) group, with 10 rats in each group. CSR model was established by inserting the wire into the cervical nerve root. The rats in the blank group were only fed normally without any intervention.The rats in the mild moxibustion group and mild moxibustion+3-MA group were given mild moxibustion at GV14 for 10 min each time,and intraperitoneal injection of 1 mL 0.9% normal saline and 1 mL 3-MA(15 mg/kg)separately. Rats in the model group were given 0.9% normal saline every day. All the three interventions were started from the 3rd day after modeling for 7 days. The rat's behavioral reaction of gait was scored and the pain threshold of rat was measured with a pain analyzer; the expressions of Beclin-1 and Bcl-2 mRNA and protein in the spinal cord (including nerve root) were detected by fluorescence quantitative PCR and immunohistochemistry, separately. The autophagosome and ultrastructure of the spinal nerve root tissue were observed by transmission electron microscope. RESULTS: After modeling, the gait score was significantly increased (P<0.01) and the pain threshold significantly decreased (P<0.01) in the model group in comparison with the blank group. There was no statistical difference in Beclin-1 and Bcl-2 mRNA and protein expression between the blank and the model groups. After intervention, compared with the model group, the gait scores were significantly reduced (P<0.01), the pain threshold and the expressions of Beclin-1 and Bcl-2 mRNA and protein were significantly increased (P<0.01，P<0.05) in the mild moxibustion and mild moxibustion+3-MA groups. The improvement of the above indicators as more significant in the mild moxibustion group than that in the mild moxibustion+3-MA group (P<0.05). After modeling, the organelles in the spinal nerve root tissue cells of the model group were damaged and there were a small amount of autophagosomes. Compared with the model group, the ultrastructure of the spinal nerve root tissue cells in the mild moxibustion group were relatively complete, and the number of autophagosomes increased. CONCLUSION: Mild moxibustion at GV14 has a good analgesic effect on CSR rats, which may be related to its effects in up-regulation of Beclin-1 and Bcl-2 expressions and activation of autophagy and inhibition of apoptosis.