Effect of Tongdu Tiaoshen electroacupuncture pretreatment on PPARγ-mediated pyroptosis of cerebral cortex in rats with cerebral ischemia reperfusion injury / 中国针灸

OBJECTIVE@#To observe the effect of Tongdu Tiaoshen (promoting the circulation of the governor vessel and regulating the spirit) electroacupuncture (EA) pretreatment on pyroptosis mediated by peroxisome proliferators-activated receptor γ (PPARγ) of the cerebral cortex in rats with cerebral ischemia reperfusion injury (CIRI) and explore the potential mechanism of EA for the prevention and treatment of CIRI.@*METHODS@#A total of 110 clean-grade male SD rats were randomly divided into a sham-operation group, a model group, an EA group, an EA + inhibitor group and an agonist group, 22 rats in each group. In the EA group, before modeling, EA was applied to "Baihui" (GV 20), "Fengfu" (GV 16) and "Dazhui" (GV 14), with disperse-dense wave, 2 Hz/5 Hz in frequency, 1 to 2 mA in intensity, lasting 20 min; once a day, consecutively for 7 days. On the base of the intervention as the EA group, on the day 7, the intraperitoneal injection with the PPARγ inhibitor, GW9662 (10 mg/kg) was delivered in the EA + inhibitor group. In the agonist group, on the day 7, the PPARγ agonist, pioglitazone hydrochloride (10 mg/kg) was injected intraperitoneally. At the end of intervention, except the sham-operation group, the modified thread embolization method was adopted to establish the right CIRI model in the rats of the other groups. Using the score of the modified neurological severity score (mNSS), the neurological defect condition of rats was evaluated. TTC staining was adopted to detect the relative cerebral infarction volume of rat, TUNEL staining was used to detect apoptosis of cerebral cortical nerve cells and the transmission electron microscope was used to observe pyroptosis of cerebral cortical neural cells. The positive expression of PPARγ and nucleotide-binding to oligomerization domain-like receptor protein 3 (NLRP3) in the cerebral cortex was detected with the immunofluorescence staining. The protein expression of PPARγ, NLRP3, cysteinyl aspartate specific protease-1 (caspase-1), gasdermin D (GSDMD) and GSDMD-N terminal (GSDMD-N) in the cerebral cortex was detected with Western blot. Using the quantitative real-time fluorescence-PCR, the mRNA expression of PPARγ, NLRP3, caspase-1 and GSDMD of the cerebral cortex was detected. The contents of interleukin (IL)-1β and IL-18 in the cerebral cortex of rats were determined by ELISA.@*RESULTS@#Compared with the sham-operation group, the mNSS, the relative cerebral infarction volume and the TUNEL positive cells rate were increased (P<0.01), pyroptosis was severe, the protein and mRNA expression levels of PPARγ, NLRP3, caspase-1 and GSDMD were elevated (P<0.01); and the protein expression of GSDMD-N and contents of IL-1β and IL-18 were increased (P<0.01) in the model group. When compared with the model group, the mNSS, the relative cerebral infarction volume and the TUNEL positive cells rate were decreased (P<0.01), pyroptosis was alleviated, the protein and mRNA expression levels of PPARγ were increased (P<0.01), the protein and mRNA expression levels of NLRP3, caspase-1 and GSDMD were decreased (P<0.01), the protein expression of GSDMD-N was reduced (P<0.01); and the contents of IL-1β and IL-18 were lower (P<0.01) in the EA group and the agonist group; while, in the EA + inhibitor group, the protein expression of PPARγ was increased (P<0.01), the protein and mRNA expression levels of NLRP3 and GSDMD were decreased (P<0.01, P<0.05), the mRNA expression of caspase-1 was reduced (P<0.01); and the contents of IL-1β and IL-18 were lower (P<0.01). When compared with the EA + inhibitor group, the mNSS, the relative cerebral infarction volume and the TUNEL positive cells rate were decreased (P<0.05, P<0.01), pyroptosis was alleviated, the protein and mRNA expression levels of PPARγ were increased (P<0.01), the protein and mRNA expression levels of NLRP3, caspase-1 and GSDMD were decreased (P<0.01), the protein expression of GSDMD-N was reduced (P<0.01); and the contents of IL-1β and IL-18 were declined (P<0.01) in the EA group. Compared with the agonist group, in the EA group, the relative cerebral infarction volume and the TUNEL positive cells rate were increased (P<0.05, P<0.01), the mRNA expression of PPARγ was decreased (P<0.01) and the protein expression of GSDMD-N was elevated (P<0.05); and the contents of IL-1β and IL-18 were higher (P<0.01).@*CONCLUSION@#Tongdu Tiaoshen EA pretreatment can attenuate the neurological impairment in the rats with CIRI, and the underlying mechanism is related to the up-regulation of PPARγ inducing the inhibition of NLRP3 in the cerebral cortex of rats so that pyroptosis is affected.

Effect of Electroacupuncture at Tinggong and Yifeng on D-galactose-induced Aging of Auditory Cortex through SIRT1/PGC-1α Pathway in Guinea Pigs / 听力学及言语疾病杂志

Objective To investigate the role of SIRT1/PGC-1α pathway in the auditory cortex aging of guinea pigs with age-related hearing loss and whether the electroacupuncture can delay aging process of the auditory cortex in the aged guinea pigs induced by D-galactose through the regulation of SIRT1/PGC-1α pathway.Methods Thirty 4-month-old guinea pigs were randomly divided into three groups including the control group (n=10),D-galactose group (model group,n =10),and D-galactose and electroacupuncture group (electroacupuncture group,n =10).The elderly group (n=10) was composed of 18-month-old guinea pigs.The guinea pigs in model group and electroacupuncture group had been subcutaneously injected with D-galactose(300 mg· kg-1· d-1)for 6 weeks.Moreover,the guinea pigs in electroacupuncture group electroacupunctured at Tinggong and Yifeng for 15 minutes half an hour later once a day.After 6 weeks,the ABR threshold of guinea pigs in each group was detected.The mRNA and protein expression of SIR1 and PG-C-1α in auditory cortex of guinea pigs were detected by real -time fluoreacence quantitative PCR and Western Blot.Results There was a significant increase in the ABR threshold of the model group and elderly group (P＜0.001,compared with the control group),decrease in the electroacupuncture group (P＜0.001,compared with the model group),and no significant difference between model group and elderly group(P＞0.05).There was significant decrease in the expression of SIRT1 and PGC-1α in the model groupand elderly group(P＜0.05,compared with the control group),and significant increase at mRNA and protein levels in the electroacupuncture group (P＜0.05,compared with the model group).Conclusion SIRT1/PG-C-1α pathway may play a role in aging process of the auditory cortex in the aged guinea pigs induced by D-galactose,and electroacupuncture at Tinggong and Yifeng may increase the antioxidant capacity of auditory cortex by activating SIRT1/PGC 1α in guinea pigs with age-related hearing loss,thus delaying the aging process of auditory cortex.