Electroacupuncture Alleviates Pain Responses and Inflammation in Collagen-Induced Arthritis Rats via Suppressing the TLR2/4-MyD88-NF-κB Signaling Pathway.

Results: EA intervention and OxPAPC injection could relieve mechanical allodynia and thermal hyperalgesia caused by CIA. Paw edema and pathological damage of synovium were significantly ameliorated after EA intervention and OxPAPC injection. Furthermore, EA intervention and OxPAPC injection markedly reduced the contents of serum TNF-α, IL-1ß, and IL-6, as well as the protein expression levels of synovial TLR2, TLR4, MyD88, and NF-κB p-p65. In particular, the expression of TLR2 and TLR4 on synovial fibroblasts and macrophages in synovium was significantly reduced by EA intervention. Conclusions: Repeated EA stimulation at ST36 and SP6 can effectively relieve joint pain and synovial inflammation caused by RA in CIA rats. The analgesic and anti-inflammatory effect of EA may be closely related to the inhibition of innate immune responses driven by the TLR2/4-MyD88-NF-κB signaling pathway in the synovium.

[Effects of transcutaneous auricular vagus nerve stimulation on bone and cartilage destruction in rats with rheumatoid arthritis].

OBJECTIVE: To observe the alleviating effect of transcutaneous auricular vagus nerve stimulation (taVNS) on articular cartilage and bone destruction in rats with collagen-induced arthritis (CIA), and explore the cellular and molecular mechanisms of taVNS against rheumatoid arthritis (RA). METHODS: The male SD rats were randomly divided into normal control group (n=12), model group (n=12), and taVNS group (n=12). The CIA rat model was established by multi-point injection of emulsion prepared from type Ⅱ bovine collagen and Freund's incomplete adjuvant into the root of rat tail. The rats in the taVNS group were treated with taVNS at bilateral auricular conchae, 30 min per time, once a day, for consecutive 28 d. The cartilage destruction of the ankle joint was observed by safranin O-fast green staining, the production of osteoclasts in the joint tissue by tartrate-resistant acid phosphatase (TRAP) staining, and the bone erosion by X-ray and Micro-CT imaging. The protein expression levels of matrix metalloproteinase (MMP)-1, MMP-3, MMP-13, receptor activator of nuclear factor-κB ligand (RANKL), and osteoprotegerin (OPG) in the synovial tissues were detected by Western blot. RESULTS: Compared with the normal control group, the CIA rats presented with typical RA symptoms and elevated arthritis index (AI,P<0.05). After intervention with taVNS, the AI remarkably declined in comparison with that in the model group (P<0.05). Compared with the control group, the model group displayed loss of cartilage matrix in the ankle joint, thinned cartilage layer, obvious cartilage damage, and increased number of osteo-clasts in the joint (P<0.01); the imaging results showed bone loss and three-dimensional structural destruction of ankle joint and aggravated bone erosion (P<0.01); the expression levels of MMP-1, MMP-3 and MMP-13, and RANKL/OPG ratio were significantly elevated in the synovial tissue of ankle joint (P<0.01, P<0.05), while the expression level of OPG was decreased (P<0.05). Compared with the model group, taVNS resulted in relatively intact cartilage layer of ankle joint, alleviated cartilage destruction, decreased number of osteoclasts (P<0.01), improved bone erosion, loss, and three-dimensional structural destruction (P<0.01), and diminished MMP-1, MMP-3, and MMP-13 expression and RANKL/OPG ratio in the synovial tissue of ankle joint (P<0.05, P<0.01), while the expression level of OPG was increased (P<0.05). CONCLUSION: taVNS effectively relieves bone and cartilage destruction in CIA rats, which might be related to its efficacy in reducing the production of osteoclasts in joint tissues and down-regulating the expression levels of MMP-1, MMP-3 and MMP-13, and RANKL/OPG ratio.

The anti-neoplastic activities of aloperine in HeLa cervical cancer cells are associated with inhibition of the IL-6-JAK1-STAT3 feedback loop.

Cervical cancer (CC) is recognized as the most common neoplasm in the female reproductive system worldwide. The lack of chemotherapeutic agents with outstanding effectiveness and safety severely compromises the anti-cipated prognosis of patients. Aloperine (ALO) is a natural quinolizidine alkaloid with marked anti-cancer effects on multiple malignancies as well as favorable activity in relieving inflammation, allergies and infection. However, its therapeutic efficacy and underlying mechanism in CC are still unclear. In the current study, MTT assay was employed to evaluate the viability of HeLa cells exposed to ALO to preliminarily estimate the effectiveness of ALO in CC. Then, the effects of ALO on the proliferation and apoptosis of HeLa cells were further investigated by plate colony formation and flow cytometry, respectively, while the migration and invasion of ALO-treated HeLa cells were evaluated using Transwell assay. Moreover, nude mice were subcutaneously inoculated with HeLa cells to demonstrate the anti-CC properties of ALO in vivo. The molecular mechanisms underlying these effects of ALO were evaluated by Western blot and immunohistochemical analysis. This study experimentally demonstrated that ALO inhibited the proliferation of HeLa cells via G2 phase cell cycle arrest. Simultaneously, ALO promoted an increase in the percentage of apoptotic HeLa cells by increasing the Bax/Bcl-2 ratio. Additionally, the migration and invasion of HeLa cells were attenuated by ALO treatment, which was considered to result from inhibition of epithelial-to-mesenchymal transition. For molecular mechanisms, the expression and activation of the IL-6-JAK1-STAT3 feedback loop were markedly suppressed by ALO treatment. This study indicated that ALO markedly suppresses the proliferation, migration and invasion and enhances the apoptosis of HeLa cells. In addition, these prominent anti-CC properties of ALO are associated with repression of the IL-6-JAK1-STAT3 feedback loop.

[Effect of electroacupuncture on articular morphological changes and serum inflammatory factors and synovial MMPs in collagen-induced arthritis rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Zusanli"(ST36) and "Sanyinjiao"(SP6) on serum TNF-α, IL-1ß, and IL-6 and expression of synovial matrimetalloproteinases (MMPs) and articular morphology in collagen-induced arthritis (CIA) rats, so as to explore its mechanisms underlying relief of arthritis. METHODS: Thirty male SD rats were randomly divided into normal control, CIA model and EA groups (n=10 rats per group). The arthritis model was induced by multi-point intradermal injection of bovine type Ⅱ collagen emulsion. EA (2 Hz/100 Hz, 1 mA) was applied to bilateral ST36 and SP6 for 30 min, once a day for 28 days. The hind-limb paw volume was measured and the arthritis index (AI) score given according to the swelling degree, rigidity and deformity of the ankle joint (0-4 points). After EA intervention, the morphological damage of the affected ankle joints was revealed by H.E. staining, safranin O-fast green staining, and tartrate-resistant acid phosphatase (TRAP) staining, separately. The levels of serum TNF-α, IL-1ß, and IL-6 were measured by ELISA, and the expression levels of MMP-1, MMP-3, MMP-13, and receptor activator of nuclear factor Kappa B ligand (RANKL) in the synovial tissue were detected by Western blot. RESULTS: Compared with the normal control group, the paw volume, AI score, TRAP-revealed number of osteoclasts, contents of serum TNF-α, IL-1ß and IL-6, and expression levels of MMP-1, MMP-3, MMP-13 and RANKL proteins were significantly increased in the model group (P<0.01, P<0.05). Following the intervention, the paw volume, AI score, number of osteoclasts, contents of serum TNF- α, IL-1ß and IL-6, and expression levels of MMP-1, MMP-3, MMP-13 and RANKL proteins were significantly decreased in the EA group (P<0.05, P<0.01) in contrast to the model group. H.E. and safranin O-fast green staining showed rough articular cartilage surface with thinned cartilage layer, obvious hyperplasia of the synovial tissue with many inflammatory cells, and serious damage and degradation of the cartilage matrix in the model group, these situations were relatively milder in the EA group. CONCLUSION: EA of ST36 and SP6 can reduce the articular damage in collagen-induced arthritis rats, which is associated with its function in reducing inflammatory response and down-regulating the expression of synovial MMP-1, MMP-3, MMP-13 and RANKL proteins.

Retrospective study of 213 cases of Stevens-Johnson syndrome and toxic epidermal necrolysis from China.

BACKGROUND: Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are rare but severe adverse drug reactions with high mortality. The use of corticosteroids and the management of complications (e.g. infection) in SJS/TEN remains controversial. METHODS: A retrospective study was performed among 213 patients with SJS/TEN who were hospitalized in our department between 2008 and 2018, to investigate the causative agents, clinical characteristics, complications, and prognoses of SJS/TEN mainly treated by systemic corticosteroids combined with intravenous immunoglobulin (IVIG). RESULTS: The causative drugs of SJS/TEN in these patients mainly consisted of antibiotics (61/213, 28.6%), anticonvulsants (52/213, 24.4%), and nonsteroidal anti-inflammation drugs (24/213, 11.3%), among which carbamazepine was the most frequently administered drug (39/213, 18.3%). There were significant differences in the maximum dosage, time to corticosteroid tapering, and the total dosage of corticosteroid between the SJS group and the TEN group, as well as among the three groups (P = 0.000), whereas in the initial dose of corticosteroid was not statistically significant among the three groups (P = 0.277). In a series of 213 cases, 18.4 cases (8.6%) were expected to die based on the score for the toxic epidermal necrolysis (SCORTEN) system, whereas eight deaths (3.8%) were observed; the difference was not statistically significant (P = 0.067; SMR = 0.43, 95% CI: 0.06, 0.48). The most common complications were electrolyte disturbance (174/213, 81.7%), drug-induced liver injury (64/213, 30.0%), infection (53/213, 24.9%), and fasting blood sugar above 10 mmol/L (33/213, 15.5%). Respiratory system (22/213, 10.3%) and wound (11/213, 5.2%) were the most common sites of infection. Multivariate logistic regression analysis indicated that the maximum blood sugar (≥10 mmol/L), the time to corticosteroid tapering (≥12 d), the maximum dosage of corticosteroid (≥1.5 mg/kg/d), and the total body surface area (TBSA) (≥10%) were defined as the most relevant factors of the infection. CONCLUSION: The mortality of patients in this study was lower than that predicted by SCORTEN, although there was no significant difference between them. Hyperglycemia, high-dose corticosteroid, and the TBSA were closely related to the infections of patients with SJS/TEN.

Contribution of GABAergic modulation in DRGs to electroacupuncture analgesia in incisional neck pain rats.

PURPOSE: Acupuncture therapy is effective for relieving postoperative pain. Our previous study showed that electroacupuncture (EA) at Futu (LI18) and Hegu (LI4)-Neiguan (PC6) could alleviate incisional neck pain, which was related with its effect in upregulating γ-aminobutyric acid (GABA) expression in cervical (C3-6) dorsal root ganglions (DRGs); but whether its receptor subsets GABAAα2R and GABABR1 in C3-6 DRGs are involved in EA analgesia or not, it remains unknown. MATERIALS AND METHODS: Seventy-five male Sprague Dawley rats were randomized to normal control, model, LI18, LI4-PC6, and Zusanli (ST36)-Yanglingquan (GB34) groups. The incisional neck pain model was established by making a longitudinal incision along the midline of the rats' neck, followed by repeated mechanical stimulation. EA was applied to bilateral LI18, LI4-PC6, or ST36-GB34 for 30 minutes at 4, 24, and 48 hours after operation. The thermal pain threshold of the neck was detected by a tail-flick unit, and the C3-6 DRGs were removed for assaying the immunoactivity of substance P (SP), GABAAα2R, glial fibrillary acidic protein (GFAP; a marker of satellite glial cells [SGCs]), and GABABR1 and the expression of GABAAα2R and GABABR1 mRNA and proteins using immunofluorescence, real-time PCR, and Western blotting, respectively. RESULTS: The cervical thermal pain threshold was significantly lower in the model group than the normal group (P<0.001), indicating hyperalgesia after neck incision, and was considerably increased in both EA-LI18 and LI4-PC6 groups (P<0.001), but not in ST36-GB34 group compared with model group (P>0.05). Immunofluorescence staining showed that GABAAα2 R expressed on SP+ neurons, and GABABR1 on SGCs. EA of LI18 and LI4-PC6 markedly suppressed the modeling-induced upregulation of the immunoactivity of SP (P<0.001 and P<0.01, respectively) and GFAP (P<0.01 and P<0.001, respectively) and significantly reversed neck incision-induced downregulation of the expression of GABAAα2R and GABABR1 mRNAs and proteins (P<0.05). CONCLUSION: EA of LI18 and LI4-PC6 has an analgesic effect in incisional neck pain rats, which is related to its effects in upregulating GABAergic inhibitory modulation on nociceptive peptidergic neurons and SGCs in cervical DRGs.

[Electroacupuncture of Multiple Acupoints Is Significantly Superior to That of Single Acupoint in Improving Sleep by Regulating Serum Sleep-promoting and Awakening-promoting Factors in Insomnia Rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation of single and multiple acupoints on sleep and concentrations of interlukin-1 ß(IL-1 ß), brain-derived neurotrophic factor (BDNF), prostaglandin D2(PGD2) and melatonin (MLT, sleep-promoting factors) and corticosterone (CORT, awakening-promoting factor) in the serum in insomnia rats, so as to explore its efficacy difference and the mechanism underlying improving sleep. METHODS: Fifty-four male SD rats were randomly divided into control, model, EA-Baihui (GV 20), EA-Shenmen (HT 7), EA-Sanyinjiao (SP 6) and EA-GV 20＋HT 7＋SP 6 groups (n＝9 rats in each group). The insomnia model was established by intraperitoneal injection of para-chlorophenylalanine (PCPA, 300 mg/kg) once daily for 2 days. In the EA-GV 20, EA-HT 7, EA-SP 6 and EA-GV 20＋HT 7＋SP 6 groups, EA stimulation was administrated for 30 min, once a day for 4 days. The sleep onset latency and sleep duration were measured after intraperitoneal injection of pentobarbital sodium (35 mg/kg). The concentrations of IL-1 ß， BDNF, MLT, PGD2and CORT in the serum were detected by ELISA. RESULTS: After EA stimulation of GV 20, HT 7, SP 6 and GV 20＋HT 7＋SP 6, the sleep latency was significantly shortened (P<0.05, P<0.01, except SP 6), and the sleep duration was remarkably prolonged in comparison with the model group (P<0.05, P<0.01), and the therapeutic effects of EA-GV 20＋HT 7＋SP 6 were significantly superior to those of EA-GV 20, EA-HT 7 and EA-SP 6 in shortening the sleep latency and lengthening the sleep duration (P<0.05). Following modeling, the concentrations of IL-1 ß， BDNF, PGD2 and MLT were significantly down-regulated, and the CORT level was markedly up-regulated in the model group relevant to the control group (P<0.05). Following EA，modeling induced dramatic decrease of serum IL-1 ß， BDNF, PGD2 and MLT was considerably up-regulated, and the increased CORT level markedly down-regulated in the EA-GV 20, EA-HT 7, EA-SP 6 and EA-GV 20＋HT 7＋SP 6 groups (P<0.05). The effects of EA-GV 20＋HT 7＋SP 6 were evidently superior to those of EA-GV 20 and EA-SP 6 in up-regulating serum IL-1 ß， BDNF and PGD2levels, and to those of HT 7, GV 20 and SP 6 in up-regulating serum MLT level, and significantly superior to those of EA-ST 7 and EA-SP 6 in down-regulating serum CORT (P<0.05). CONCLUSION: EA stimulation of HT 7, GV 20, SP 6 and GV 20＋HT 7＋ SP 6 can significantly improve the sleep in insomnia rats, which is closely associated with its effects in regulating serum sleep-promoting factors and awakening-promoting factor. Joint administration of EA of GV 20＋HT 7＋ SP 6 has a better effect than the single acupoint mentioned above.

[Transcutaneous Electrostimulation of Auricular Otopoints Reduces Epileptic Attack Possibly by Suppressing Hippocampal Gliocyte Proliferation and Regulating IL-6 and IL-10 Expression in Chronic Temporal Lobe Epilepsy Rats].

OBJECTIVE: To observe the effect of transcutaneous otopoint electrostimulaiton (TCOES) on seizure frequency, immunoreactivity of hippocampal gliocytes and expression of proinflammatory cytokine interleukin-6(IL-6) and anti-inflammatory cytokine IL-10 in chronic temporal lobe epilepsy (CTLE) rats, so as to investigate its antiepileptic mechanism. METHODS: Thirty-six SD rats were randomly divided into control, model and TCOES groups (n=12 in each group). The CTLE model was established by intraperitoneal injection (i.p.i.) of lithium chloride (127.2 mg/kg), scopolamine (1 mg/kg, 20 h after the 1st injection) and pilocarpine (10 mg/kg, 30 min after scopolamine injection). Rats of the control group were treated by i.p.i. of normal saline. TCOES (1 mA, 20 Hz) was applied to bilateral otopoint "Heart"-"Lung"-"Subcortex" region for 20 min, once daily for 6 weeks. The epileptic attack was observed by a video monitoring system. The numbers of ionized calcium-binding adapter molecule-1 (Iba 1)-labeled microgliacytes and glial fibrillary acidic protein (GFAP)-labeled astrocytes in the CA 1 and CA 3 regions of hippocampus were counted under light microscope after immunostaining, and the expression levels of hippocampal IL-6 and IL-10 proteins and genes were determined by immunofluorescence and quantitative real-time PCR, respectively. RESULTS: After TCOES intervention, the seizure frequency was significantly decreased in comparison with pre-treatment(P<0.05), modeling-induced dramatic increase of the numbers of microgliacytes and astrocytes,IL-6 immunoactivity in the hippocampal CA 1 and CA 3 regions, and IL-6 mRNA expression in the hippocampus were significantly suppressed (P<0.05), and hippo-campal IL-10 immunoactivity and mRNA expression were considerably up-regulated in comparison with the model group (P<0.05). CONCLUSIONS: TCOES intervention has an antiepileptic effect in CTLE rats, which may be associated with its effects in suppressing gliocyte proliferation, suppressing the expression of proinflammatory cytokine IL-6, and up-regulaiting the expression of anti-inflammatory cytokine IL-10 in the hippocampus.

[Effects of Electroacupuncture on Activities of Satellite Glial Cells of Dorsal Root Ganglia in Rats with Neck Incision Pain].

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation of "Futu"(LI 18), etc. on activities of satellite glial cells (SGCs) in the dorsal root ganglia (DRG) in rats with neck-incision pain so as to explore its mechanism underlying reduction of post-surgical pain of thyroidectomy. METHODS: Male SD rats were randomly divided into control, model, EA-Futu (LI 18), EA-Hegu (LI 4)-Neiguan (PC 6), and EA-Zusanli (ST 36)-Yanglingquan (GB 34) groups, with 20 rats in each group. The neck-incision pain model was established by making a longitudinal incision and repeated mechanical stimulation. In the EA-LI 18, EA-LI 4-PC 6 and EA-ST 36-GB 34 groups, EA stimulation was administrated for 30 min, once a day,continuously for 3 days. The thermal pain threshold (PT) of the neck-incision region was detected. The immunoactivity of glial fibrillary acidic protein (GFAP,a specific marker for SGCs) and connexin 43 (Cx 43) of DRGs (C 2-C 6) was determined by fluorescent immunohistochemistry. The expression levels of GFAP, IL-1 ß, IL-6, and TNF-α mRNAs were determined by quantitative Real-time PCR, and the contents of IL-1 ß,IL-6,TNF-α assayed by enzyme linked immunosorbent assay (ELISA) and the expression of Cx 43 protein was detected by Western blot. RESULTS: After EA intervention at LI 18 and LI 4-PC 6 (but not ST 36-GB 34), neck incision-induced reduction of the thermal PT was obviously prolonged in comparison with the model group (P<0.05),suggesting a pain relief. The expression levels of GFAP, IL-1 ß, IL-6 and TNF-α mRNAs and Cx 43 protein, and the contents of IL-1 ß, IL-6 and TNF-α in C 2-C 6 DRGs were all significantly up-regulated in the model group relevant to those of the control group (P<0.05). Following EA, modeling induced dramatic increase of expression of GFAP, IL-1 ß, IL-6 and TNF-α mRNAs and Cx 43 protein in both EA-LI 18 and EA-LI 4-PC 6 groups, and the contents of IL-1 ß and TNF-α in the EA-LI 18 group, IL-6 in the EA-LI 4-PC 6 group was considerably down-regulated (P<0.05). In comparison with the model group, no significant changes were found in all the abovementioned indexes of EA-ST 36 -GB 34 group except the down-regulated IL-1 ß and TNF-α mRNAs, in the contents of IL-1 ß and TNF-α of the EA-LI 4-PC 6 group, and in the IL-6 content of the EA-LI 18 group (P>0.05). CONCLUSIONS: EA stimulation of LI 18 and LI 4-PC 6 can significantly suppress pain reaction of neck incision in the rat, which is closely associated with its effects in down-regulating the activity of SGCs, decreasing the release of pro-inflammatory cytokines and in weakening the expression of Cx 43 in the cervical DRGs.

Effect of electroacupuncture on thermal pain threshold and expression of calcitonin-gene related peptide, substance P and γ-aminobutyric acid in the cervical dorsal root ganglion of rats with incisional neck pain.

OBJECTIVE: Acupuncture therapy effectively reduces post-surgical pain, but its mechanism of action remains unclear. The aim of this study was to investigate whether expression of γ-aminobutyric acid (GABA) and the neuropeptides substance P (SP) and calcitonin gene-related peptide (CGRP) in the primary sensory neurons of cervical dorsal root ganglia (DRG) are involved in electroacupuncture (EA)-induced analgesia in a rat model of incisional neck pain. METHODS: The pain model was established by making a longitudinal midline neck incision in 60 rats. Another 15 rats underwent sham surgery (normal group). Post-incision, 15 rats remained untreated (model group) and 45 rats underwent EA (frequency 2/100 Hz, intensity 1 mA) at bilateral LI18, LI4-PC6 or ST36-GB34 (n=15 each) for 30 min at 4 hours, 24 hours, and 48 hours post-surgery, followed by thermal pain threshold (PT) measurement. 30 min later, the rats were euthanased and cervical (C3-6) DRGs removed for measurement of immunoreactivity and mRNA expression of SP/CGRP and the GABAergic neuronal marker glutamic acid decarboxylase 67 (GAD67). RESULTS: Thermal PT was significantly lower in the model group versus the normal group and increased in the LI18 and LI4-PC6 groups but not the ST36-GB34 group compared with the model group. Additionally, EA at LI18 and LI4-PC6 markedly suppressed neck incision-induced upregulation of mRNA/protein expression of SP/CGRP, and upregulated mRNA/protein expression of GAD67 in the DRGs of C3-6 segments. CONCLUSIONS: EA at LI18/LI4-PC6 increases PT in rats with incisional neck pain, which is likely related to downregulation of pronociceptive mediators SP/CGRP and upregulation of the inhibitory transmitter GABA in the primary sensory neurons of cervical DRGs.

[Effects of Transcutaneous Electrostimulation of Auricular Points on Behavior and Hippocampal IL-1 ß and TNF-α Expression in Temporal Lobe Epilepsy Rats].

OBJECTIVE: To observe the effect of transcutaneous otopoint electrostimulation (TCOES) on behavior and expression of hippocampal interleukin-1 ß (IL-1 ß) and tumor necrosis factor-α (TNF-α) expression in lithium-pilocarpine induced chronic spontaneous temporal lobe epilepsy (TLE) rats, so as to investigate its antiepileptic mechanism. METHODS: Thirty-six SD rats were randomly divided into control, model and TCOES groups (n=12 in each group). The epilepsy model was established by intraperitoneal injection of lithium chloride (127.2 mg/kg), scopolamine(1 mg/kg, 20 h after the 1st injection) and pilocarpine (10 mg/kg, 30 min after scopolamine injection). Rats of the control group were treated by injection of normal saline(i.p.i.). Transcutaneous electrostimulation (1 mA, 20 Hz) was applied to bilateral otopoints "Heart" "Lung" and "Subcortex" for 20 min, once daily for 6 weeks except the weekends. The behavior reactions were observed by a video monitoring system. The expression of IL-1 ß and TNF-α proteins and genes in the hippocampus were determined by immunofluorescence and quantitative real-time PCR, separately. RESULTS: Behavioral observation showed that after TCOES intervention, the frequency of epilepsy attack was significantly decreased in comparison with pre-treatment (P<0.05). Immunofluorescence and real-time PCR showed that compared with the control group, the immunoactivity of IL-1 ß and TNF-α in both hippocampal CA 1 and CA 3 regions and hippocampal IL-1 ß and TNF-α gene expression were obviously increased in the model group (P<0.05, P<0.01). Following TCOES, the increased hippocampal IL-1 ß and TNF-α and IL-1 ß mRNA and TNF-α mRNA expression levels were all suppressed (P<0.05, P<0.01). CONCLUSIONS: TCOES intervention has an antiepileptogenic effect in temporal lobe epilepsy rats, which may be related to its effects in down-regulating expression of proinflammatory cytokine IL-1 ß and TNF-α in the hippocampus.

Effect of Electroacupuncture Intervention on Expression of CGRP, SP, COX-1, and PGE2 of Dorsal Portion of the Cervical Spinal Cord in Rats with Neck-Incision Pain.

The present study was aimed to determine if cervicospinal substance P (SP) and its neurokinin-1 receptor (NK-1R), calcitonin gene-related peptide (CGRP), cyclooxygenase-1 (COX-1), and prostaglandin E2 (PGE2) were involved in electroacupuncture (EA) analgesia in neck-incision pain rats. EA intervention was applied to bilateral Futu (LI18), Hegu (LI4)-Neiguan (PC6), and Zusanli (ST36)-Yanglingquan (GB34) for 30 min. Cervicospinal SP and CGRP immunoactivity was detected by immunofluorescence technique, NK-1R and COX-1 protein and mRNA expression levels were determined using Western blot and real-time PCR, respectively, and PGE2 content was measured using ELISA. Outcomes indicated that EA of EA-LI18 and LI4-PC6 (not ST36-GB34) significantly suppressed neck-incision induced decrease of thermal pain threshold (P < 0.05). EA stimulation of LI18 and LI4-PC6 markedly inhibited neck-incision induced upregulation of SP and CGRP immunoactivity, NK-1 R and COX-1 mRNA and protein expression levels, as well as the increase of PGE2 content in the dorsal cervicospinal cord (P < 0.05). These findings showed that LI18 and LI4-PC6 EA stimulation-induced downregulation of SP, CGRP, NK-1R, COX-1, and PGE2 levels in the dorsal cervicospinal cord may contribute to their effects in relieving neck-incision pain. This study highlights the targets of EA intervention for reducing post-thyroid-surgery pain for the first time.

[Influence of electroacupuncture on efficacy of estrogen in regulating hypothalamic reproductive endocrine activity in rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the efficacy of estrogen-induced regulation of reproductive endocrine in ovariectomized (OVX) rats. METHODS: Thirty SD rats were randomly divided into control, model (OVX), EA-Guanyuan(CV 4), Estrogen (E) and EA+ E groups (n = 6 in each group). OVX model was duplicated by removing the bilateral ovaries. EA (5 Hz/20 Hz, 1- 2 mA) was applied to "Guanyuan" (CV 4) for 30 min, once every other day and for 3 sessions. Estrogen at doses of 0.5 mg/kg and 0. 25 mg/kg was intraperitonealy given to rats in the E group and EA+ E group, respectively. Serum estrogen and cortisol contents were assayed by enzyme linked immunosorbent assay and the expression of hypothalamic gonadotrophin releasing hormone(GnRH)mRNA, G protein-coupled receptor 54(GPR 54)mRNA, kisspeptin-1 (Kiss-l) mRNA and corticotropin releasing hormone (CRH) mRNA were detected by using real time polymerase chain reaction (RT-PCR). RESULTS: Compared with the control group, the expression levels of GnRH mRNA, GPR 54 mRNA and Kiss-1 mRNA in hypothalamus in the model group were increased significantly (P<0. 05), while the expression of hypothalamic CRH mRNA was down-regulated obviously (P<0. 05), and serum cortisol and estrogen levels were reduced significantly (P<0. 05). In comparison with the model group, the expression levels of hypothalamic GnRH mRNA in the EA, E and EA+ E groups, and those of hypothalamic GPR 54 mRNA and Kiss-1 mRNA in the E and EA+ E groups were down-regulated considerably (P<0. 05); whereas hypothalamic CRH mRNA expression levels in the E and EA+ E groups were up-regulated obviously (P<0. 05),and serum cortisol and estrogen contents in the E and EA+ E groups were increased remarkably (P<0.05). Comparison among the EA and EA+ E groups showed that the effects of EA+ E group were significantly superior to those of EA group in down-regulating hypothalalmic GnRH mRNA and GPR 54 mRNA expression, and in up-regulating hypothalamic CRH mRNA expression and serum cortisol and estrogen contents (P<0.05). No significant differences were found between E and EA + E groups in down-regulating hypothalamic GnRH mRNA, GPR 54 mRNA and Kiss-1 mRNA expression, and in up-regulating hypothalamic CRH mRNA expression and serum cortisol and estrogen levels (P>0. 05). CONCLUSION: EA-CV 4 is able to potentiate the effects of low-dose of estrogen in down-regulating hypothalamic GnRH mRNA,GPR 54 mRNA and Kiss-1 mRNA expression, and up-regulating hypothalamic CRH mRNA expression and raising serum cortisol and estrogen contents in OVX rats, showing a favorable modulation effect on reproductive endocrine activity.

[Effects of electroacupuncture at "Futu" (LI 18), etc. on expression of spinal 5-HT 1 AR mRNA, 5-HT 2 AR mRNA and protein in rats with neck incision pain].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at bilateral "Futu" (LI 18), etc. on the expression of 5-HT 1 A receptor (R) mRNA, 5-HT2 AR mRNA and protein in the spinal cord of rats with neck incision pain, so as to explore its underlying mechanism in relieving incision pain. METHODS: A total of 48 Wistar rats were randomly divided into control, model (incision pain), Futu (LI 18), Hegu (LI 4)-Neiguan (PC 6, LI 4-PC 6), Zusanli (ST 36)-Yanglingquan (GB 34) cervical cord (ST 36-GB 34 C) and lumbar cord (ST 36-GB 34 L) groups. A 1.5 cm longitudinal incision was made in the middle of the neck under Isoflurane inhalation anesthesia. Pain threshold (PT) was measured using radiant heat. EA (1-2 mA, 2 Hz/15 Hz) was applied to bilateral LI 18, LI 4-PC 6, ST 36-GB 34 for 30 min. The expression of 5-HT 1 AR mRNA, 5-HT 2 AR mRNA and protein in the cervical spinal cord (C1-C4) tissue, and 5-HT 1 AR mRNA and 5-HT 2 AR mRNA in the lumbar cord (L1- L3) were detected with Real-time PCR and Western blot analysis, respectively. RESULTS: After the operation, the thermal PT was shortened obviously In comparison with pre-operation (P < 0.05). Compared with post-operation, the PT values were increased markedly in the LI 18 group and LI 4-PC 6 group (P < 0.05), rather than in the ST 36-GB 34 group (P > 0.05). The expression levels of 5-HT 1 AR mRNA, 5-HT 2 AR mRNA and 5-HT 2 AR protein in the cervical cord of the model group were increased significantly compared with those of the control group (P < 0.05). In comparison with the model group, the expression level of 5-HT 1 AR mRNA was down-regulated considerably in the LI 18 and LI 4-PC 6 groups (P < 0.05), and those of spinal 5-HT 2 AR mRNA and protein were up-regulated significantly in the LI 18 and LI 4-PC 6 groups (P < 0.05). No significant changes were found in the expression of 5-HT 1 AR mRNA and 5-HT 2 AR mRNA in the ST 36-GB 34 C group in comparison with the model group (P > 0.05). The expression levels of 5-HT 1 AR mRNA and 5-HT 2 AR mRNA in the ST 36-GB 34 L group were significantly lower than those of the ST 36-GB 34 C group (P < 0.05). CONCLUSION: EA of LI 18 and LI 4-PC 6 can significantly suppress pain reaction of neck incision in the rat, which is closely associated with its effects in down-regulating the expression of 5-HT 1 AR mRNA and in up-regulating 5-HT 2 AR mRNA and 5-HT 2 AR protein in the cervical spinal cord.

[Effects of electroacupuncture at "Futu" (LI 18) on the immunoactivity of substance P, 5-HT 1 AR, etc. of the cervical spinal dorsal horn in rats with neck incision pain].

OBJECTIVE: To observe the effect of electroacupuncture(EA) of bilateral "Futu" (LI 18) on the immunoactivity of substance P (SP), NK-1 receptor (R), cyclooxygenase-1 (COX-1), 5-HT 1 AR and 5-HT2 AR in the cervical spinal dorsal horn of rats with neck incision pain, so as to explore its underlying mechanism in relieving incision pain. METHODS: A total of 40 Wistar rats were randomly divided into control, model (incision pain), EA-Futu(LI 18), and EA-Zusanli(ST 36)-Yanglingquan (GB 34, EA-ST 36-GB 34) groups (n = 10/group). A 1.5 cm longitudinal incision was made along the middle of the neck in pentobarbital-anesthetized rats. Pain threshold (PT) was measured using radiant heat. EA (1-2 mA, 2 Hz/15 Hz) was applied to bilateral LI 18, ST 36-GB 34 for 30 min. The expression of SP, NK-1 R, COX-1, 5-HT 1 AR and 5-HT2 AR in the spinal cord (C1-C4) tissue were detected with immunohistochemistry. RESULTS: After the operation, the thermal PT was decreased obviously in markedly after treatment (P < 0.05), and no significant changes were found in PT in EA-ST 36-GB 34 group (P > 0.05). The immunoactivity of SP, NK-1 R, COX-1 in the superficial dorsal horn in model group was increased significantly and that of 5-HT 1 AR and 5-HT 2 AR significantly and mildly, separately compared with control group (P < 0.05). In comparison with model group, the immunoreactivity of SP, NK-1 R and COX-1 was decreased considerably, and that of 5-HT 1 AR and 5-HT 2 AR was upregulated remarkably in EA-LI 18 group (P < 0.05). No significant changes were found in the immunoactivity of SP, NK-1 R, COX-1, 5-HT 1 AR and 5-HT 2 AR in EA-ST 36-GB 34 group (P > 0.05). CONCLUSION: EA-Futu (LI 18) can significantly suppress pain reaction of neck incision in the rat, which is closely associated with its effects in downregulating the expression of SP, NK-1 R, COX-1 and upregulating that of 5-HT 1 AR and 5-HT 2 AR in the spinal dorsal horn.

[Application of molecular biological techniques to the study on acupuncture and moxibustion].

In the present paper, the authors introduce recent progress of application of molecular biological techniques in the studies on the underlying mechanism of acupuncture therapy for 1) resisting inflammation and relieving pain, 2) resisting tumors, 3) improving ischemic cerebral and myocardial injury, and 4) delaying aging from gene transcription and expression (synthesis of mRNA); and for relieving sciatic nerve injury, spinal traumatic injury, and cerebral ischemia from protein expression or phosphorylation. In addition, the authors also make a preliminary analysis on the extant problems of the application of techniques of both genomics and proteomics in acupuncture research at the present and predict their applicable prospect.