Sirtuin 5 alleviates apoptosis and autophagy stimulated by ammonium chloride in bovine mammary epithelial cells.

Ammonia (NH3) is an irritating and harmful gas that affects cell apoptosis and autophagy. Sirtuin 5 (SIRT5) has multiple enzymatic activities and regulates NH3-induced autophagy in tumor cells. In order to determine whether SIRT5 regulates NH3-induced bovine mammary epithelial cell apoptosis and autophagy, cells with SIRT5 overexpression or knockdown were generated and in addition, bovine mammary epithelial cells were treated with SIRT5 inhibitors. The results showed that SIRT5 overexpression reduced the content of NH3 and glutamate in cells by inhibiting glutaminase activity in glutamine metabolism, and reduced the ratio of ADP/ATP. The results in the SIRT5 knockdown and inhibitor groups were comparable, including increased content of NH3 and glutamate in cells by activating glutaminase activity, and an elevated ratio of ADP/ATP. It was further confirmed that SIRT5 inhibited the apoptosis and autophagy of bovine mammary epithelial cells through reverse transcription-quantitative PCR, western blot, flow cytometry with Annexin V FITC/PI staining and transmission electron microscopy. In addition, it was also found that the addition of LY294002 or Rapamycin inhibited the PI3K/Akt or mTOR kinase signal, decreasing the apoptosis and autophagy activities of bovine mammary epithelial cells induced by SIRT5-inhibited NH3. In summary, the PI3K/Akt/mTOR signal involved in NH3-induced cell autophagy and apoptosis relies on the regulation of SIRT5. This study provides a new theory for the use of NH3 to regulate bovine mammary epithelial cell apoptosis and autophagy, and provides guidance for improving the health and production performance of dairy cows.

Performance of the Improved Priestley-Taylor Model for Simulating Evapotranspiration of Greenhouse Tomato at Different Growth Stages.

Mastering crop evapotranspiration (ET) and improving the accuracy of ET simulation is critical for optimizing the irrigation schedule and saving water resources, particularly for crops cultivated in a greenhouse. Taking greenhouse-grown tomato under drip irrigation as an example, two weighing lysimeters were used to monitor ET at two seasons (2019 and 2020), whilst meteorological factors inside the greenhouse were measured using an automatic weather station. Then the path analysis approach was employed to determine the main environmental control factors of ET. On this basis, an improved Priestley-Taylor (IPT) model was developed to simulate tomato ET at different growth stages by considering the influence of environmental changes on model parameters (e.g., leaf senescence coefficient, temperature constraint coefficient and soil evaporative water stress coefficient). Results showed that the average daily ET varied from 0.06 to 6.57 mm d−1, which were ~0.98, ~2.58, ~3.70 and ~3.32 mm/d at the initial, development, middle and late stages, respectively, with the total ET over the whole growth stage of ~333.0 mm. Net solar radiation (Rn) and vapor pressure deficit (VPD) were the direct influencing factors of ET, whereas air temperature (Ta) was the limiting factor and wind speed (u2) had a little influence on ET. The order of correlation coefficients between meteorological factors and ET at two seasons was Rn > VPD > Ta > u2. The IPT model can accurately simulate ET in hourly and daily scales. The root mean square error of hourly ET at four stages changed from 0.002 to 0.08 mm h−1 and daily ET varied from 0.54 to 0.57 mm d−1. The IPT coefficient was close to the recommended PT coefficient (1.26) when the average Ta approaches 26 °C and LAI approaches 2.5 cm2 cm−2 in greenhouse conditions. Our results can provide a theoretical basis for further optimization of greenhouse crop irrigation schedules and improvement of water use efficiency.

Effect of a Superabsorbent Polymer (Poly-Gamma-Glutamic Acid) on Water and Salt Transport in Saline Soils under the Influence of Multiple Factors.

In order to effectively suppress the negative effects of salt ions contained in saline soils on agricultural soil quality and crop growth, this study took advantage of the water-saving properties and better soil improvement properties of poly-γ-glutamic acid (γ-PGA). By carrying out various experiments, the following relationships have been found. (1) The lab experiment studies the effect of the γ-PGA application on the infiltration of sandy loam soil. The application rates of γ-PGA are 0%, 0.1%, 0.2%, and 0.3%, respectively. (2) HYDRUS-1D is used to simulate water infiltration of sandy loam soil under multiple factors (bulk density, γ-PGA application rate, and the application depth of γ-PGA). (3) The effect of γ-PGA on soil solute (Cl−) transport is also explored in this paper. The results show that bulk density and the application depth of γ-PGA (p < 0.01) have higher effects on cumulative infiltration than the application amount of γ-PGA (p < 0.05). A lower γ-PGA application rate will increase the proportion of unavailable soil water by 3%. The established empirical models have good results. Furthermore, when the γ-PGA application rate is 0.3% (0.02-cm2 min−1), the Cl− hydrodynamic dispersion coefficient is the highest. The study recommends applying the γ-PGA at 1.4 g cm−3, 5−20 cm, and 0.2%. The results of this study are conducive to an in-depth understanding of the physicochemical properties of poly-γ-glutamic acid, improving the utilization rate of salinized land, achieving agricultural water and fertilizer conservation and yield enhancement, and guaranteeing sustainable land use and sustainable development of agroecological environment.

Morus alba leaves ethanol extract protects pancreatic islet cells against dysfunction and death by inducing autophagy in type 2 diabetes.

BACKGROUND: Protection of pancreatic islet cells against dysfunction or death by regulating autophagy is considered to be an effective method for treatment of type 2 diabetes mellitus (T2DM). Morus alba leaves (mulberry leaves), a popular herbal medicine, have been used for prevention of T2DM since ancient times. PURPOSE: This study aimed to clarify whether Morus alba leaves ethanol extract (MLE) could protect islet cells in vivo and in vitro by regulating autophagy in T2DM, and explore the possible mechanism of action. METHODS: The main chemical constituents in MLE were analyzed by HPLC. The T2DM rat model was induced via high-fat diet combined with peritoneal injection of low-dose streptozotocin, and MLE was administered by oral gavage. Fasting blood glucose (FBG) and plasma insulin were measured, and homeostatic model assessment of ß cell function (HOMA-ß) and insulin resistance (HOMA-IR) were determined. The histomorphology of pancreas islets was evaluated by haematoxylin and eosin staining. In palmitic acid (PA)-stressed INS-1 rat insulinoma cells, cell viability was assayed by an MTT method. Expression of the autophagy-related proteins LC3 I/II, p62, p-AMPK and p-mTOR in islet tissues and INS-1 cells was evaluated by western blotting or immunohistochemistry analysis. RESULTS: The four main chemical constituents in MLE were identified as chlorogenic acid, rutin, isoquercitrin and quercitrin. MLE ameliorated hyperglycemia, insulin resistance and dyslipidemia of T2DM rats with prominent therapeutic effect. Further study indicated that MLE observably improved islet function, alleviated islet injury of T2DM rats, and inhibited PA-induced INS-1 cell death. On the other hand, MLE significantly induced autophagy in islet cells both in vivo and in vitro, and autophagy inhibitors abolished its therapeutic effect on T2DM rats and protective effect on islet cells. Apart from this, MLE markedly activated the AMPK/mTOR pathway in INS-1 cells, and the AMPK inhibitor prevented the autophagy induction ability of MLE. CONCLUSION: Together, MLE could protect islet cells against dysfunction and death by inducing AMPK/mTOR-mediated autophagy in T2DM, and these findings provide a new perspective for understanding the treatment mechanism of Morus alba leaves against T2DM.

P450 enzymes-based metabolic interactions between monarch drugs and the other constituent herbs: A strategy to explore compatibility mechanism of Sangju-Yin.

BACKGROUND: Herbal compatibility of compound formulas can enhance therapeutic effects or reduce side effects of the monarch drugs, but majority of compatibility mechanisms are still unknown. Sangju-Yin, a well-known Chinese compound formula, is currently used to treat common cold in clinical. PURPOSE: In this study, we proposed a strategy to explore the compatibility mechanism of Sangju-Yin by investigating P450 enzymes-based metabolic interactions between monarch drugs and the other constituent herbs. METHODS: Under the guidance of traditional Chinese medicine theory, the constituent herbs of Sangju-Yin were divided into four groups, including monarch drugs, monarch drugs with addition of minister drugs, monarch drugs with addition of minister and adjuvant drugs, as well as the whole recipe, namely monarch drugs with addition of minister, adjuvant and conductant drugs. Their effects on rats in vivo P450 (CYP1A2, CYP2A3, CYP2C6, CYP2C11 and CYP3A1) activities after oral administration were evaluated using probe drug assay based on LC-MS/MS. Moreover, effects of the four groups of herbs on mRNA expression of P450 enzymes after oral administration, as well as in vitro P450 activities after co-incubation, were investigated to explore the underlying mechanisms. RESULTS: Comparing with monarch drugs, addition of different constituent herbs significantly enhanced CYP1A2 and CYP2C6 activities, and inhibited CYP2A3 and CYP3A1 activities, indicating their possible influences on plasma concentrations of active constituents in the monarch drugs. Mechanism study suggested that these herbs affected P450 activities by transcriptional regulation and/or direct interaction with the enzymes. CONCLUSION: This study clarified the compatibility mechanism of Sangju-Yin from the aspect of P450 enzymes-based metabolic interactions, which would benefit better understanding of the therapeutic basis of Sangju-Yin.