[Tetramethylpyrazine regulates angiogenesis of endothelial cells in cerebral ischemic stroke injury via SIRT1/VEGFA signaling pathway].

This study aims to investigate whether tetramethylpyrazine(TMP) can stimulate angiogenesis in cerebral microvascular endothelial cells and alleviate cerebral ischemic stroke(CIS) and to explore the underlying mechanisms. In the animal study, adult Sprague-Dawley rats(n=15) were assigned into sham surgery(sham), middle cerebral artery occlusion/reperfusion(MCAO/R), and MCAO/R+TMP(intraperitoneal injection of 20 mg·kg~(-1)) groups. The neurological function was evaluated by the Z-Longa method. The cerebral infarction volume was detected by TTC staining. Enzyme-linked immunosorbent assay(ELISA) was employed to detect the expression of vascular endothelial growth factor(VEGF), angiopoietin(Ang), and platelet-derived growth factor(PDGF). Immunofluorescence staining was employed to detect Ki67 and the expression of vascular endothelial growth factor A(VEGFA) and slient information regulator 1(SIRT1). Western blot was employed to determine the expression levels of VEGFA, SIRT1, angiopoietin-2(Ang-2), and platelet-derived growth factor B(PDGFB). In the cell study, mouse brain-derived endothelial cells(Bend.3) were cultured, and the optimal concentration of TMP was determined. Then, VEGF, Ang, and PDGF were detected by ELISA after the addition of cabozantinib. Western blot was employed to measure the expression of VEGFA, Ang-2, and PDGFB. Immunofluorescence staining was used to detect CD31, CD34, and Ki67, and the proliferation, migration, and tube formation ability of Bend.3 cells were observed in vitro. Western blot and immunofluorescence staining were performed to measure the expression of SIRT1 and VEGFA after addition of the SIRT1-specific inhibitor selisistat(EX-527). The results showed that compared with the sham group, the MCAO/R group had severe neurological function damage, increased infarction volume, up-regulated expression of VEGF, VEGFA, Ang, Ang-2, PDGF, and PDGFB, and down-regulated expression of Ki67 and SIRT1(P<0.01). Compared with the MCAO/R group, the MCAO/R+TMP group presented alleviated neurological function damage, reduced infarction volume, and activated expression of VEGF, VEGFA, Ang, Ang-2, PDGF, PDGFB, Ki67, and SIRT1(P<0.01). The cell experiments showed that compared with the normal group, Bend.3 cells were activated by oxygen glucose deprivation/reoxygenation(OGD/R) treatment(P<0.05, P<0.01). Compared with the OGD/R group, the OGD/R+TMP group upregulated the expression levels of VEGF, VEGFA, Ang, Ang-2, PDGF, PDGFB, SIRT1, Ki67, CD31, and CD34, enhanced the angiogenic ability of Bend.3 cells without being inhibited by BMS or EX-527(P<0.05, P<0.01, P<0.001). The results suggest that TMP can activate the SIRT1/VEGFA signaling pathway to stimulate angiogenesis and alleviate CIS injury.

[Acupuncture ameliorates neurological function by suppressing microglia polarization and inflammatory response after cerebral ischemia in rats].

OBJECTIVE: To observe the effect of acupuncture on microglia polarization and inflammatory reaction in rats with cerebral ischemia-reperfusion injury (CIRI), so as to explore its mechanisms underlying improvement of CIRI. METHODS: Thirty male SD rats were randomly divided into sham operation, model, and acupuncture groups, with 10 rats in each group. The CIRI model was established by occlusion of the middle cerebral artery (MCAO) for 1 h, followed by reperfusion. After modeling, rats in the acupuncture group received manual acupuncture stimulation of "Dazhui" (GV14), "Baihui"(GV20), "Shuigou" (GV26), bilateral "Zusanli" (ST36) and "Fengchi" (GB20) by twirling the needles rapidly for 10 s/acupoint every 10 min, with the needles retained for 20 min. The treatment was conducted once daily for successive 7 days. The neurological function was evaluated according to Longa's method. The state of CIRI was observed after Nissl staining, and the expression levels of Iba-1, iNOS, Arg1, BDNF, GDNF and NeuN in the ischemic cortex tissue were detected by immunofluorescence staining. The contents of TNF-α, IL-6 and IL-10 in the ischemic tissue were assayed by ELISA. The protein expression levels of BDNF, GDNF, TLR4, MyD88 and NF-κB in the ischemic tissues were detected by Western blot. RESULTS: The neurological deficit score on the 24 h and 7th day was considerably higher in the model group than in the sham operation group (P<0.01), and evidently lower on the 7th day in the acupuncture group than in the model group (P<0.01). The number of NeuN positive cells，the area of immunofluorescence dual labelling of Arg1, BDNF and GDNF positive staining, IL-10 content, BDNF and GDNF protein expressions were significantly decreased (P<0.01), and the immunofluorescence dual labelling area of Iba-1 and iNOS, TNF-α and IL-6 contents, the pretein expression levels of TLR4, MyD88 and NF-κB considerably increased (P<0.01) in the model group relevant to the sham operation group. In contrast to the model group, the acupuncture group had a significant increase in the number of NeuN positive cells, the immunofluorescence dual labelling area of Arg1, BDNF and GDNF positive staining, IL-10 content, and BDNF and GDNF protein expressions (P<0.05, P<0.01), and an evident decrease in Iba-1 and iNOS positive staining, contents of TNF-α and IL-6, and the protein expression levels of TLR4, MyD88 and NF-κB (P<0.01, P<0.05). Nissl staining showed a marked reduction in the number of neurons, the nucleus pyknosis and nissl bodies and loose arrangement of the neuronal cells in the model group, which was relatively milder in the acupuncture group. CONCLUSION: Acupuncture intervention can improve neurological function in CIRI rats, which may be related to its effects in regulating the polarization of microglia, reducing inflammatory reaction and increasing the secretion of neurotrophic factors in the brain, inhibiting TLR4/MyD88/NF-κB signaling pathway.

Wuzi Yanzong pill attenuates MPTP-induced Parkinson's Disease via PI3K/Akt signaling pathway.

Wuzi Yanzong Pill (WYP) was found to play a protective role on nerve cells and neurological diseases, however the molecular mechanism is unclear. To understand the molecular mechanisms that underly the neuroprotective effect of WYP on dopaminergic neurons in Parkinson's disease (PD). PD mouse model was induced by the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Gait and hanging tests were used to assess motor behavioral function. Immunofluorescence assay was used to determine TH-positive neurons in substantia nigra (SN). Apoptosis, dopamine and neurotrophic factors as well as expression of PI3K/Akt pathway were detected by TUNEL staining, ELISA and western blotting, respectively. First, it was observed that WYP intervention improved abnormal motor function in MPTP-induced PD model, alleviated the loss of TH+ neurons in SN, and increased dopamine content in brain, revealing a potential protective effect. Second, network pharmacology was used to analyze the possible targets and pathways of WYP action in the treatment of PD. A total of 126 active components related to PD were screened in WYP, and the related core targets included ALB, GAPDH, Akt1, TP53, IL6 and TNF. Particularly, the effect of WYP on PD may be medicate through PI3K/Akt signaling pathway and apoptotic regulation. The WYP treated PD mice had higher expression of p-PI3K, p-Akt and Bcl-2 but lower expression of Bax and cleaved caspase-3 than the non-WYP treated PD mice. Secretion of brain-derived neurotrophic factor (BDNF) and cerebral dopamine neurotrophic factor (CDNF) were also increased in the treated mice. WYP may inhibit apoptosis and increase the secretion of neurotrophic factor via activating PI3K/ Akt signaling pathway, thus protecting the loss of dopamine neurons in MPTP-induced PD mice.

[Response of understory species distribution of Robinia pseudoacacia plantation to environmental factors in loess hilly region, China.] / é»„åœŸä¸˜é™µåŒºåˆºæ§ç¾¤è½æž—ä¸‹ç‰©ç§åˆ†å¸ƒå¯¹çŽ¯å¢ƒå› å­çš„å“åº”.

We explored the distribution characteristics and influencing factors of understory species in Robinia pseudoacacia plantation in Wuqi, Ansai, Mizhi, Yichuan and other places, in combination with niche breadth, soil, altitude and other environmental factors. We analyzed the response mechanism of species distribution to environmental factor to provide a scientific basis for the mana-gement of R. pseudoacacia plantation in loess hilly region. The results showed that the broadly distributed understory species in R. pseudoacacia plantation were Setaria viridis, Heteropappus altaicus, Artemisia scoparia, Poa sphondylodes, Artemisia leucophylla, Ixeris sonchifolia, and Incarvillea sinensis. With the increasing rehabilitationage, the dominant understory species distribution followed the order: Artemisia capillaries → P. sphondylodes → A. scoparia → others (Rubia cordifolia, Rosa rubus and so on) → A. leucophylla → S. viridis. Results from principal component analysis showed that soil total phosphorus (25.6%), altitude (20.3%) and soil total nitrogen (19.3%) were the key factors influencing understory species distribution in R. pseudoacacia plantation. Soil organic carbon content, soil total nitrogen content, soil total phosphorus content, soil water content and the number of understory species distribution in R. pseudoacacia plantation were generally positively correlated with the degree of correlation varying across different species. There was no correlation between the slope aspect and the understory species distribution in R. pseudoacacia plantation. In conclusion, topography and soil factors played important roles in driving understory species distribution. The steeper the slope, the higher the altitude, the fewer understory species in R. pseudoacacia plantation. Understory species distribution was mainly affected by soil total phosphorus content and altitude. The understory species distribution reflected the differences of soil nutrient status, which had a certain guiding role in the management of R. pseudoacacia plantation.

[Stoichiometric characteristics of Artemisia sacrorum communities under different site conditions and their correlation with soil nutrients]. / ä¸åŒç«‹åœ°é“æ†è’¿åŒ–å­¦è®¡é‡ç‰¹å¾åŠå ¶ä¸ŽåœŸå£¤å »åˆ†çš„å ³ç³».

We explored the stoichiometric characteristic of Artemisia sacrorum communities and its correlation with soil characters by considering the aspect and slope position, and variation of soil nutrient in Yangqingchuan catchment of Wuqi. Our results showed that total carbon, total nitrogen, total phosphorus contents and C:N ratio in A. sacrorum (both shoot and root) in the top of hillock>sunny slope>semi-shady slope>shady slope. The C:P ratio in shoot and the N:P ratio in root decreased consistently. The N:P ratio in shoot and the C:P ratio in root decreased first and then increased. While the total nitrogen and total phosphorus in shoot, and the organic carbon in root increased first and then decreased with the decreases of slope position, the C:N ratio and N:P ratio in shoot decreased first and then increased. The stoichiometric characteristic of A. sacrorum communities was positively correlated with the soil stoichiometric characteristics. However, the C:N ratio, C:P ratio and N:P ratio of A. sacrorum and the total phosphorus of root were negatively correlated to corresponding soil indices. The correlation between shoot nutrition and soil was greater than that between soil and root nutrition. In conclusion, middle position of shady slope was optimal for the growth of A. sacrorum. The stoichiometric characteristics of plants were significantly correlated with soil nutrient condition. Aspect and slope position had a significant effect on the stoichiometric cha-racteristics of A. sacrorum community and soil. Proper A. sacrorum community could help restore soil nutrition.

[Variation in leaf functional traits of different-aged Robinia pseudoacacia communities and relationships with soil nutrients]. / ä¸åŒæž—é¾„åˆºæ§å¶åŠŸèƒ½æ€§çŠ¶å·®å¼‚åŠå ¶ä¸ŽåœŸå£¤å »åˆ†çš„å ³ç³».

On the basis of various leaf functional traits of different-aged Robinia pseudoacacia communities, as well as the relationships between the traits and soil nutrients, the adaptation strategy of R. pseudoacacia in relation to soil conditions was analyzed in Ansai County, the Loess Plateau, China. The results showed that specific leaf area, leaf area, leaf water content, leaf total nitrogen content and leaf organic carbon content first increased and then decreased with the increasing stand age. The peak values of specific leaf area (279.18 cm2·g-1), leaf area (12.33 cm2), leaf water content (0.09%), leaf total nitrogen content (33.01 g·kg-1) and leaf organic carbon content (523.08 g·kg-1) were obtained at 30 years old. With the increasing stand age, leaf tissue density, leaf total phosphorus content, leaf thickness and stomata density increased, and stomata length and stomata width decreased. Leaf area, specific leaf area, leaf total phosphorus content, leaf water content, leaf thicknessand stomata density were the main indexes based on principal component analysis (PCA), for R. pseudoacacia leaf functional traits responding to the increased stand age. The indexes were correlated with each other, indicating that R. pseudoacacia was capable of changing leaf morphological structure to adapt to environmental changes. Soil total nitrogen content was the main factor influencing leaf area, leaf water content, specific leaf area and stomata length, while soil organic carbon content mainly affected stomata width, leaf tissue density, leaf thickness, leaf total phosphorus content, leaf total nitrogen content and stomata density. Therefore, soil total nitrogen and organic carbon content were main factors that affected leaf functional traits of R. pseu-doacacia in different stand ages. Soil nutrients in R. pseudoacacia communities were improved with the increasing stand age, which eventually affected leaf functional traits. The flexibility of leaf functional traits indicated that R. pseudoacacia communities had great potential to adapt to environmental change in Loess Plateau hilly region.