[Electroacupuncture pretreatment alleviates apoptosis of renal tubular epithelia cells by down-regulating expression of TRPC6 in hyperglycemic mice].

OBJECTIVE: To observe the effect of electroacupuncture (EA) pretreatment on apoptosis of renal tubular epithelial cells in mice with hyperglycemia, so as to explore its mechanisms underlying protecting the kidney from hyperglycemia-induced injury. METHODS: Eighty male C57BL/6 mice were equally and randomly divided into control, model, EA and sham EA groups. The hyperglycemia model was established by intraperitoneal injection of streptozocin (STZ, 50 mg·kg-1·d-1) for 5 consecutive days. Before modeling, EA (2 Hz/15 Hz, 0.3-0.5 mA) was applied to bilateral "Zusanli" (ST36) and "Shenshu" (BL23) for 30 min, once daily for 7 days, while mice in the sham EA group were treated with the same acupoints but without electrical stimulation. The blood glucose values were measured after fasting for 6 hours after 3 days of modeling. The degree of renal tissue injury was observed by microscope after H.E. staining, and the apoptosis level of renal tubular epithelial cells observed by TUNEL staining. The expression levels of transient receptor potential channel 6 (TRPC6) and related apoptotic proteins Caspase-3, Bax and Bcl-2 in the renal tissue were detected by Western blot and immunohistochemistry, separately. RESULTS: Compared with the control group, the blood glucose content and the expression levels of TRPC6, Caspase-3 and Bax proteins, as well as the level of the renal apoptotic cells were significantly increased (P<0.001, P<0.000 1), while the expression level of Bcl-2 protein and the ratio of Bcl-2/Bax were remarkably decreased in the model group (P<0.000 1). In comparison with the model and sham EA groups, the blood glucose content, percentage of apoptotic cells and the expression levels of TRPC6, Caspase-3 and Bax were significantly decreased (P<0.01, P<0.000 1, P<0.05, P<0.001), and the expression level of Bcl-2 and the ratio of Bcl-2/Bax were apparently increased in the EA group (P<0.01, P<0.05, P<0.001). HE statin showed abnormal dilation of the capillary lumen and disappearance of the proximal tubules in the model group, which was relatively milder in the EA group. CONCLUSION: EA pretreatment can lower blood glucose level and reduce renal apoptosis in hyperglycemia mice, which may be related to its effects in down-regulating the expression of TRPC6 and Caspase-3 and up-regulating the ratio of Bcl-2/Bax.

Phragmunis a suppresses glioblastoma through the regulation of MCL1-FBXW7 by blocking ELK1-SRF complex-dependent transcription.

Glioblastoma (GBM) is a highly aggressive brain tumor. During screening work, we found a new compound named phragmunis A (PGA), which is derived from the fruitbody of Trogia venenata, exhibits a potential cytotoxic effect on patient-derived recurrent GBM cells and temozolomide (TMZ)-resistant cell lines. The present study was designed to investigate the potential molecular mechanism of the anti-glioma effects of PGA in vitro and in vivo. Studies investigating the mechanism revealed that PGA diminished the binding efficiency of ETS family of transcription factor (ELK1) and Serum response factor (SRF), and suppressed ELK1-SRF complex-dependent transcription, which decreased the transcriptional levels of downstream genes Early growth response protein 1 (EGR1)-Polycomb ring finger (BMI1), thus inducing the imbalanced regulation between Myeloid cell leukaemia-1 (MCL1) and F-Box and WD repeat domain containing 7 (FBXW7). Finally, orthotopic xenograft models were established to confirm the anti-glioma effect of PGA on tumour growth. We showed, for the first time, that the cytotoxic effects of PGA occurred by inducing MCL1 inhibition and FBXW7 activation by blocking ELK1-SRF complex-dependent transcription. The blockage of ELK1-mediated transcription resulted in the suppression of EGR1-BMI1, which led to the upregulation of FBXW7 expression and downregulation of MCL1. These findings suggested that PGA could be a therapeutic drug candidate for the treatment of recurrent GBM by targeting the ELK1-SRF complex.

[Electroacupuncture preconditioning alleviates kidney injury in hyperglycemic mice].

OBJECTIVE: To explore the protective effect and molecular mechanism of electroacupuncture (EA) preconditioning on renal injury induced by streptozotocin (STZ) in hyperglycemic mice. METHODS: Eighty male C57BL/6 mice were randomly divided into 4 groups: control, model, EA and sham EA groups, n=20 in each group. The hyperglycemic model was established by intraperitoneal injection of 0.1% STZ solution (50 mg/kg) for 5 days. EA preconditioning or sham EA was applied at "Zusanli" (ST36) and "Shenshu" (BL23), once daily for 7 successive days in the EA or sham EA group. Three days after mode-ling, the blood glucose was measured after fasting for 6 hours. The degree of renal injury was observed by HE staining and PAS staining; the expressions of transient receptor potential cation channel 6 (TRPC6) and Nephrin protein in glomerulus were observed by immunohistochemistry; the expressions of TRPC6 and Nephrin protein in renal cortex were detected by Western blot. RESULTS: Compared with the control group, the blood glucose level was significantly increased (P<0.01), the glomerular cross-sectional area was apparently increased (P<0.000 1), the glomerular capillaries dilated, the matrix proliferated, the brush edge of renal tubules disappeared, the proportion of abnormal renal tubules increased (P<0.000 1), and the expressions of TRPC6 and Nephrin in glomeruli and renal tissue were significantly increased (P<0.01) in the model group. Following EA treatment and compared with the model and sham EA groups, the blood glucose was significantly down-regulated (P<0.01), the renal tissue injury was apparently alleviate, the cross-sectional area of glomerulus was reduced (P<0.05), the brush edge of renal tubules changes were obviously improved, the proportion of abnormal renal tubules decreased (P<0.000 1), and the expressions of TRPC6 and Nephrin in glomerulus and renal tissue decreased (P<0.05) in the EA group. CONCLUSION: EA preconditioning can alleviate renal injury in hyperglycemic mice, which is closely related to its effects in reducing renal TRPC6 and Nephrin expressions and inhibiting podocyte activation.

High-Tech Acupuncture for Prevention of Lifestyle Diseases-A Sino-Austrian Cooperation Research Protocol on Heart Rate Variability.

BACKGROUND: Acupuncture can not only be used for well-known diseases, but also for so-called modern lifestyle-related diseases. Using innovative methods like e.g. new analyses of heart rate variability (HRV), it is also possible to investigate diseases like burnout syndrome, ie., qi deficiency in Chinese medicine (CM). OBJECTIVE: The main object of this research protocol is to perform research on the relationship of burnout syndrome and heart rate (HR) and HRV. METHODS: A total of 175 patients with burnout syndrome (qi deficiency syndrome) in five groups and 35 healthy volunteers will be investigated. Based on random numbers generated by computer and concealed in opaque envelops, the patients will be assigned to four acupuncture groups using Zusanli (ST 36) acupuncture stimulation, Guanyuan (CV4) acupuncture stimulation, both points, and both points with Streitberger device respectively, and a moxibustion group using both points mentioned above, with 35 cases in each group. Altogether four different experiments are planned. Experiment 1 includes 70 subjects and is a comparison between a burnout group and a control group (healthy volunteers). The evaluation parameters are different scores and indices of HR and HRV. Experiment 2 includes 140 subjects and compares the efficacy of different acupuncture points. In experiment 3 (105 subjects), acupuncture and moxibustion should be compared to healthy volunteers. Experiment 4 (70 subjects) investigates the long-term therapeutic effects of acupuncture and moxibustion on the scores of qi deficiency and HR/HRV in qi deficiency patients. In both the acupuncture and moxibustion groups, a total of 10 treatments will be performed. CONCLUSIONS: The joint research aims at the scientific evaluation of CM, mainly in the field of HRV. This parameter could be a very good indicator of the state of health and can be inflfluenced by different acupuncture methods, as shown in the past.

[Comparison of Protective Effect of Electroacupuncture on Myocardial Ischemia Injury Between Different Acupoint Formulas in Rats].

OBJECTIVE: To observe the different effects of electroacupucnture of "Neiguan" (PC 6) grouping with other acupoints on myocardial ischemia (MI) injury, so as to provide evidence for clinical acupoint formulas. METHODS: Forty Wistar male rats were randomly divided into normal, model, EA 1[bilateral "Neiguan" (PC 6), bilateral "Zusanli" (ST 36),"Guanyuan" (CV 4)], and EA 2[bilateral "Neiguan" (PC 6), bilateral "Shenmen" (HT 7), "Danzhong" (CV 17)] groups, 10 rats in each group. The MI model was established by subcutaneous injection with ISO solution(5 mL/kg), once per day for 7 days. Both EA 1 and EA 2 groups were subjected to EA (2 Hz/100 Hz, 1 mA) for 10 min before subcutaneous injection, once daily for 21 days. Electrocardiogram (ECG) was recorded after treatment. Serum cTnT was detected by biochemistry. SOD, GSH-Px and MDA in myocardial tissues of left ventricular were measured by ELISA, and the ultrastructural changes of myocardial cells in the left ventricular were observed by electron microscope. RESULTS: Compared with the normal group, the amplitude of S-T segment and HR of ECG, cTnT and MDA contents were significantly increased in the model group (P<0.01) accompanied with seriously destroyed ultrastructure in the myocardial tissue, while SOD and GSH-Px levels were lower than those in the normal group (P<0.01). However, EA treatments reversed all the abnormal changes in the model group (P<0.05). There was no significant difference between EA 1 and EA 2 groups in the amplitude of S-T segment and HR (P>0.05); cTnT and MDA levels in the EA 1 group were lower than those in the EA 2 group (P<0.05), while SOD and GSH-Px levels were higher than those in the EA 2 group (P<0.05), accompanied with lighter degree of ultrastructure damage in the myocardial tissue in the EA 1 group. CONCLUSIONS: EA can prevent myocardial ischemic injury, and the effect of EA 1 treatment was better than that of EA 2.

[Effects of electroacupuncture at "Neiguan" (PC 6) on p38 MAPK signaling pathway in rats with cardiac hypertrophy].

OBJECTIVE: To discuss the mechanisms of electroacupuncture at "Neiguan" (PC 6) on p38 MAPK signaling pathway in rats with cardiac hypertrophy. METHODS: Forty SD rats were randomly divided into four groups: a normal group, a model group, a model plus p38 MAPK inhibitor group, a model plus electroacupuncture group, ten rats in each group. The model rats were established by subcutaneous injection 3 mg/(kg x d) of Isoprenaline Hydrochloride; model plus p38 MAPK inhibitor group were injected 0.3 mg/(kg x d) of specific inhibitor SB 203580; model plus electroacupuncture group was treated by electroacupuncture at "Neiguan"(PC 6) with continuous-wave, 2 Hz and 1 mA for 20 minutes, once a day for 14 days. There was no treatment in other two groups. The contents of TNF-alpha and IL-1beta in heart tissue were detected by radioimmunoassay and the p38 MAPK, p-p38 MAPK by western blot. RESULTS: Compared with normal group, the contents of TNF-alpha, IL-1beta, p38 MAPK, p-p38 MAPK were significantly increased in model group (all P < 0.01). The contents of TNFalpha, IL-1beta, p38 MAPK, p-p38 MAPK were significantly decreased in model plus p38 MAPK inhibitor group and model plus electroacupuncture group, compared with model group, all P < 0.05; compared with normal group, P < 0.05, P < 0.01; but no significant difference between model plus p38 MAPK inhibitor group and model plus electroacupuncture group (P > 0.05). CONCLUSION: Electroacupuncture at "Neiguan" (PC 6) can prevent the phosphorylation of p38 MAPK of myocardial hypertrophy, and the mechanism maybe adjust p38 MAPK signaling pathway by inhibiting the expression of TNF-alpha and IL-1beta.

[Influence of electroacupuncture of Guanyuan (GV 4) and Zusanli (ST 36) on the immune function of T cells in aging rats].

OBJECTIVE: To observe the influence of electroacupuncture (EA) on T cell immunologic function in the aged rats so as to reveal its underlying mechanism in anti-aging effect. METHODS: Fifty SD rats (half male and half female) were randomly divided into normal control (young rats), aging model (model), EA group, immunosuppression (IS), and EA+ IS groups. Aging model (in the late 4 groups) was duplicated by subcutaneous injection of D-galactose for 40 days. Guanyuan (CV 4) and bilateral Zusanli (ST 36) were punctured and stimulated electrically (2 Hz, 1 mA), 15 min every time, six times a week and for four weeks. IS model was induced by i. p. i. of cytoxan (60 mg/kg) after 26 and 27 days' EA treatment. Splenetic T lymphocyte proliferation index was assayed by Methylthiazolyldiphenyl-tetrazolium bromide (MTT) chromatometry. IL-2/IL-2 receptor (R) expression level of T cells was measured by enzyme linked immunosorbent assay (ELISA). CD8+ CD28(+) -T cell subset fluorescence intensity was detected by flow cytometry. RESULTS: Compared with normal control group, the splenetic T lymphocyte proliferation index, IL-2 and IL-2R expression levels, and CD8+ CD28(+) -T cell density in model and IS groups decreased significantly (P < 0.01). In comparison with model group, the abovementioned indexes in EA and EA + IS groups increased significantly (P < 0.01), and those indexes levels of EA + IS group were also significantly higher than those in IS group ( P < 0.05, P < 0.01). Comparison between EA and EA+ IS groups showed that CD8+ CD2+ T cell expression of the later group was significantly lower than that in EA group (P < 0 05). No significant differences were found between EA and EA + IS groups in T cell proliferation rate and IL-2 and IL-2R expression levels (P > 0.05) CONCLUSION: EA of CV 4 and ST 36 induced upregulation of T cell proliferation and IL-2/IL-2R, and CD8+ CD(+)28 -T cell may contribute to its antiaging and immuno-enhancement effects in the aged rats.

[Effect of acupuncture on expression of neurology-related genes in the brain tissue of the rat].

OBJECTIVE: To analyze and explore the molecular mechanism of acupuncture precondition (AP) in improving brain ischemia reperfusion injury. METHODS: Six male Wistar rats were divided into a normal group of 3 rats and an acupuncture group of 3 rats. In the acupuncture group, acupuncture precondition was first given at "Shenshu" (BL 23) and "Baihui" (GV 20). The brain of the rat was removed 0.5 hour after the needle was withdrawn, and then gene differential expression was analyzed by genechip method. RESULTS: After acupuncture precondition for 0.5 h, expression of 265 genes showed changes of more than 2 times, and that of 20 genes had a significant difference as compared with that of the normal rat (P<0.05). Among the genes, expression of 8 was upregulated and 12 downregulated; 11 were related with neuron signal conduction (ion channel, transmitter, receptor, second message), 5 with gene transcription and control, 2 with metabolism enzyme, 1 with heat shock reaction and 1 with cell skeleton. CONCLUSION: Some gene expression substances induced by acupuncture play an important role in AP resisting brain ischemia reperfusion injury; and functional changes of neurons after AP are possibly the important basis of its subsequent action in resisting brain ischemia reperfusion injury.