Effects of electroacupuncture on mitophagy mediated by SIRT3/PINK1/Parkin pathway in Parkinson's disease mice. / ç”µé’ˆå¯¹å¸•é‡‘æ£®ç— å°é¼ SIRT3/PINK1/Parkin通路介导的线粒体自噬的影响.

OBJECTIVES: To observe the effects of electroacupuncture (EA) at "Fengfu"(GV16), "Taichong"(LR3), and "Zusanli"(ST36) on mitophagy mediated by silencing regulatory protein 3 (SIRT3)/ PTEN induced putative kinase 1 (PINK1)/PARK2 gene coding protein (Parkin) in the midbrain substantia nigra of Parkinson's disease (PD) mice, and to explore the potential mechanisms of EA in treating PD. METHODS: C57BL/6 mice were randomly divided into the control, model, EA, and sham EA groups, with 12 mice in each group. The PD mouse model was established by intraperitoneal injection of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP). The EA group received EA stimulation at GV16, LR3 and ST36, while the sham EA group received shallow needling 1 mm away from the above acupoints without electrical stimulation. The motor ability of mice in each group was evaluated using an open field experiment. Immunohistochemistry was used to detect the expression of tyrosine hydroxylase (TH) and α-synuclein (α-syn) in the substantia nigra of mice. The ultrastructure of neurons in substantia nigra was observed by transmission electron microscope (TEM). Immunofluorescence was used to detect the expression of the autophagy marker autophagy-associated protein light chain 3 (LC3). The expression levels of TH, α-syn, SIRT3, PINK1, Parkin, P62, Beclin-1, LC3Ⅱ mRNA and protein were detected by PCR and Western blot. RESULTS: Compared with the control group, mice in the model group showed a decrease in the total exercise distance, time, movement speed and times of crossing central region (P<0.01)；the positive expressions of TH and LC3 were decreased (P<0.01), while the positive expression of α-syn increased (P<0.01), accompanied by mitochondrial swelling, mitochondrial cristae fragmentation and decrease, and decreased lysosome count；the expression levels of TH, SIRT3, PINK1, Parkin, Beclin-1, and LC3Ⅱ mRNA and protein in the midbrain substantia nigra were decreased (P<0.01), while the expression levels of α-syn and P62 mRNA and protein were increased (P<0.01, P<0.05). Compared with the model group, the mice in EA group showed a significant increase in the total exercise distance, time, movement speed and times of crossing central region (P<0.01, P<0.05)；the positive expressions of TH and LC3 were increased (P<0.01, P<0.05), while the positive expression of α-syn was decreased (P<0.01), accompanied by an increase in mitochondrial count, appearance of autophagic va-cuoles, and a decrease in swelling, the expression levels of TH, SIRT3, PINK1, Parkin, Beclin-1 and LC3Ⅱ mRNA and protein in the midbrain substantia nigra were increased (P<0.01, P<0.05), while the mRNA and protein expression levels of α-syn and P62 were decreased (P<0.01)；the sham EA group showed an increase in the total exercise distance and time(P<0.05), with an increase in the positive expression of TH (P<0.05) and a decrease in the positive expression of α-syn (P<0.05)；some mitochondria exhibited swelling, and no autophagic vacuoles were observed；the protein expression levels of TH, SIRT3, Parkin and LC3Ⅱ were increased (P<0.01, P<0.05), and the expression levels of P62 mRNA, α-syn mRNA and protein were decreased (P<0.01, P<0.05), and LC3Ⅱ mRNA expression was increased (P<0.05). In comparison to the sham EA group, the EA group showed an extension in the total exercise time (P<0.01), the positive expression and mRNA expression levels of α-syn were decreased (P<0.01, P<0.05), while the expression levels of TH, SIRT3, PINK1, Parkin mRNA and SIRT3 protein were increased (P<0.05). CONCLUSIONS: EA at GV16, LR3, and ST36 can exert neuroprotective function and improve the motor ability of PD mice by activating the SIRT3/PINK1/Parkin pathway to enhance the expression of TH and reduce α-syn aggregation in the substantia nigra of PD mice.

ATPdock: a template-based method for ATP-specific protein-ligand docking.

MOTIVATION: Accurately identifying protein-ATP binding poses is significantly valuable for both basic structure biology and drug discovery. Although many docking methods have been designed, most of them require a user-defined binding site and are difficult to achieve a high-quality protein-ATP docking result. It is critical to develop a protein-ATP-specific blind docking method without user-defined binding sites. RESULTS: Here, we present ATPdock, a template-based method for docking ATP into protein. For each query protein, if no pocket site is given, ATPdock first identifies its most potential pocket using ATPbind, an ATP-binding site predictor; then, the template pocket, which is most similar to the given or identified pocket, is searched from the database of pocket-ligand structures using APoc, a pocket structural alignment tool; thirdly, the rough docking pose of ATP (rdATP) is generated using LS-align, a ligand structural alignment tool, to align the initial ATP pose to the template ligand corresponding to template pocket; finally, the Metropolis Monte Carlo simulation is used to fine-tune the rdATP under the guidance of AutoDock Vina energy function. Benchmark tests show that ATPdock significantly outperforms other state-of-the-art methods in docking accuracy. AVAILABILITY AND IMPLEMENTATION: https://jun-csbio.github.io/atpdock/. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Active components alignment of Gegenqinlian decoction protects ulcerative colitis by attenuating inflammatory and oxidative stress.

ETHNOPHARMACOLOGICAL RELEVANCE: Gegenqinlian Decoction (GQD) has been used as a folk remedy for gastrointestinal diseases in China over thousands of years. It has significant treatment efficacy for patients with inflammatory bowel disease (IBD). We analyzed and showed that the active components alignment of Gegenqinlian Decoction (ACAG) possesses broad pharmacological effects including analgesic, antipyretic, anti-inflammatory, antibacterial, antiviral and antidiarrhea, as well as the effect of adjusting gastrointestinal function in our preliminary experiments. However, the exact molecular mechanisms on how ACAG exerts these pharmacological effects still remain elusive. In the present study, the plausible pharmacological effects of ACAG on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis were investigated. MATERIALS AND METHODS: Male Sprague-Dawley (SD) rats with TNBS/ethanol-induced colitis were used. The colonic wet weight, macroscopic and histological colon injury, superoxide dismutase (SOD), malonyldialdehyde (MDA), and inducible nitric oxide synthase (iNOS) activity were observed. Pro-inflammation cytokines were determined by ELISA methods, semi-quantitative RT-PCR and Immuno-histochemistry. RESULTS: We showed administration of ACAG was able to improve colitis. This was manifested by a decreased in the score of macroscopic and histological colonic injury, by lowered colonic wet weight, accompanied by significant increased of SOD activity, and decreased of MDA and iNOS activities. The treatment also significantly reduced tumor necrosis factor-alpha (TNF-α) and interleukin-1ß (IL-1ß) levels in colon and serum as well as the colonic mRNA levels for several inflammatory cytokines such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), macrophage inflammatory protein-2 (MIP-2), intercellular adhesion molecule-1 (ICAM-1) and toll-like receptor 2, 4 (TLR2, TLR4). In addition, we also showed that ACAG was able to inhibit the activation and translocation of transcription factors, nuclear factor kappaBp65 (NF-κBp65) in colon. CONCLUSIONS: Our results suggest that ACAG exhibits protective effect in TNBS-induced ulcerative colitis. We postulate that this might be due to its modulation of oxidant/anti-oxidant balance, downregulation of productions, expressions of pro-inflammatory cytokines and inhibition of NF-κBp65 signal transduction pathways.

Neuroprotective effects of Psoralea corylifolia Linn seed extracts on mitochondrial dysfunction induced by 3-nitropropionic acid.

BACKGROUND: Mitochondrial dysfunction has been implicated in neuronal apoptosis associated with neurodegenerative diseases such as Huntington's disease (HD). Animals that are administered 3-nitropropionic acid (3-NP), a mitochondrial toxin that specifically inhibits complex II of the mitochondrial electron transport chain, manifest HD-like symptoms. METHODS: Psoralea corylifolia Linn seed extracts against 3-NP induced mitochondrial dysfunction in cultured rat pheochromocytoma (PC12) cells, which are used for neurobiological studies. RESULTS: In this study showed that 3-NP-treated PC12 cells had decreased ATP levels, lower cellular oxygen consumption, and reduced mitochondrial membrane potential than those of untreated PC12 cells. Psoralea corylifolia Linn seed extracts stimulated mitochondrial respiration with uncoupling and induced an increased bioenergetic reserve capacity. Furthermore, PC12 cells pretreated with P. corylifolia Linn seed extracts significantly attenuated 3-NP-induced cell death, reduced ATP levels, and lowered the mitochondrial membrane potential. CONCLUSIONS: These results demonstrate that P. corylifolia Linn seed extracts have a significant protective effect against 3-NP induced cytotoxicity. Thus, our results indicate that P. corylifolia Linn seed extracts may have potential applications as therapeutic agents for treating neurodegenerative disease.

[Study on the identification of radix scutellariae and extract using Fourier transform infrared spectroscopy and two-dimensional IR correlation spectroscopy].

2D-IR correlation spectroscopy was used to do the research on crude and prepared drug of radix scutellariae and the extracts of them. The results show that the holistic shape of peaks among them are similar in the FTIR spectra. In second derivative spectra, the two absorption peaks: 1,745 and 1,411 cm(-1) of processed products move to the bigger wavenumber direction, while 1,357 cm(-1) of processed products moves to the smaller wavenumber direction; There are conspicuous differences in Two-dimensional infrared correlation spectroscopy among them: Four characteristic peaks are shown between 1,300 and 1,800 cm(-1). The intensity of peak at 1,575 cm(-1) is the strongest. There are three main districts about the autopeaks of sliced scutellariae. Wine-fried scutellariae has two auto-peak districts, in which all the auto-peaks are positively correlated. The FTIR spectra of total glycoside extract of different samples present characteristic peaks at 1,615, 1,585, 1,450 cm(-1) (vibration of phenyl framework) and 1,658 cm(-1) (=C-O ) respectively, therefore, the authors speculated that their mutual component is the compound of phenolic glycoside. The two-dimensional infrared correlation spectra present five automatic peaks (vibration of phenyl framework) in 800-1,800 cm(-1) (1,366, 1,420, 1,508, 1,585, 1,669 cm(-1)). So the authors can conclude that a lot of information can be provided by macro-fingerprint technology of infrared spectroscopy which can evaluate overall quality of radix scutellariae accurately and be used to study the characteristics of relevance of crude and prepared scutellariae.

Study on similar traditional Chinese medicines cornu Cervi pantotrichum, cornu Cervi and cornu Cervi degelatinatum by FT-IR and 2D-IR correlation spectroscopy.

As complicated mixture systems, traditional Chinese medicines (TCM) are very difficult to identify and discriminate, especially for the drug samples have the same source. Therefore, finding a rapid and effective identification method for the similar TCM is the most pressing task in TCM research. In this paper, the macroscopic IR fingerprint method including Fourier transform infrared spectroscopy (FT-IR), the second derivative spectra and two-dimensional correlation infrared spectroscopy (2D-IR), were applied to studying and identifying three very similar traditional Chinese animal drugs Cornu Cervi Pantotrichum (CCP), Cornu Cervi (CC) and Cornu Cervi Degelatinatum (CCD). On the basis of the different FT-IR and 2D-IR spectra ratios of amide I bands, amide II bands and Ca(3)(PO(4))(2), three similar animal drugs of the same source were successfully identified and discriminated. Then the FT-IR spectra data of total 30 samples, comprising 10 samples each of above three drugs, were obtained and analyzed with the soft independent modeling of class analogy (SIMCA) pattern recognition technique. The result showed that the three kinds of drug samples located in three different clusters separately. As a result, it was demonstrated that the macroscopic IR fingerprint method could discriminate the three similar TCM, CCP, CC and CCD, rapidly, effectively and non-destructively.

[Study on active components alignment of alkaloids in extractive of Folium aconiti kusnezoffii by ESI-MS].

OBJECTIVE: To analyze the active components alignment of aconitum alkaloids on extractive of Folium Aconiti Kusnezoffii and its decoction. METHOD: The analysis was performed by the method of ESI-MS. RESULT: The biester-type and the triester-type diterpenoid aconitum alkaloids were hydrolysed after boiling in water, the former was hydrolyzed to 14-benzoylaconine and aconine, the latter to 3-acylaconine. CONCLUSION: The method is simple rapid and sensitive, provide a new instrumentality to analyze the active components alignment of aconitum alkaloids on extractive of folium aconiti kusnezoffii.

[FTIR and classification study on the Chinese compound recipe].

On the basis of building a model of main components and with the fingerprint infrared spectrum as the object-Xiaoerchoufeng powders were rapidly classified and closely studied by using the method of clustering analysis. Because of the tiny difference between the spectra of different compound recipes, including the compound recipe without scolopendra, the compound recipe without scorpio, the compound recipe without bombyx batryticatus, the compound recipe without eupolyphaga seu steleophaga and the compound recipe without periostracum cryptotympanae, the outcome of the clustering is ideal. When tested with the blind samples (which were picked out from the chosen samples) the accuracy of clustering reaches up to 90%. In order to increase the clustering accuracy, the number of the samples should be increased and also these samples should be more characteristic. On the whole, with the combination of the fingerprint spectrum and the clustering analysis, the compound recipe can be distinguished rapidly.

[Identification study of different Flos Trollii by IR].

OBJECTIVE: To establish a method to identify different Flos Trollii with IR spectra and to provide a new technique for their science estimation and quality identification. METHODS: Their IR spectra were obtained by the method of intuitionistic analysis-selecting point to normalization. RESULTS: Differences on the shape, the position, the height and the relative intensity of absorption peaks were shown in IR spectra of the ten samples. CONCLUSIONS: Different Flos Trollii can be identified using IR spectra obtained by the method of intuitionistic analysis-selecting point to normalization. This new method can be used to study the identification features of the influence factors of Traditional Chinese Medicine quality.