【Objective】 To investigate the possible mechanism of sacubitril valsartan sodium (LCZ696) and valsartan in protecting rat cardiomyocytes under diabetic cardiomyopathy (DCM) by observing their effects on CGRP, TGF-β1/Smad7, caspase-3/Bcl-2 and Fas/FasL signaling pathways. 【Methods】 The diabetic model was made by the method of high glucose and high-fat diet combined with intraperitoneal injection of streptozotocin (STZ). Two rats were killed at the 4th, 6th and 8th week, respectively, to observe the myocardial structure. The myocardial structure of the rats changed at the 8th week, which was regarded as the success construction of diabetic cardiomyopathy model. The rats were randomly divided into four groups: control+alcohol group, DCM+alcohol group, DCM+valsartan (DCM+VST) group, and DCM+LCZ696 (DCM+SVST) group. After that, equivalently intense alcohol was given to control+alcohol group and DCM+alcohol group, and 30 mg/(kg·d) valsartan was given to DCM+VST group, and 60 mg/(kg·d) LZC696 given to DCM+SVST group. At the end of the experiment, HE and Masson staining were used to observe the changes of myocardial histopathology; TUNEL used to observe the changes of myocardial apoptosis; immunohistochemistry used to detect the expressions of TGF-β1/ Smad7 and CGRP protein, Western blotting used to detect the expressions of caspase-3, Bcl-2, and Fas/FasL protein. 【Results】 Compared with those in control+alcohol group, rats in DCM+alcohol group had significantly decreased cardiac LVEF and LVFS values (P<0.05); cardiomyocyte hypertrophy; malalignment; more obvious interstitial fibrosis; more myocyte apoptosis (P<0.05); increased TGF-β1, caspase-3, and Fas/FasL protein expressions; and decreased CGRP, Smad7, and Bcl-2 protein expressions (P<0.05). Compared with DCM+alcohol group, DCM+VST group and DCM+SVST group ended up with improved rat cardiac function indicators and pathological changes; decreased TGF-β1, caspase-3 and Fas/FasL protein expressions(P<0.05); and increased CGRP, Smad7 and Bcl-2 protein expressions (P<0.05); DCM+SVST group was even superior to DCM+VST group in these aspects(P<0.05). 【Conclusion】 LCZ696 can antagonize the heart injury of DCM more effectively than valsartan. It protects the myocardium by regulating TGF-β1/Smad7, caspase-3/Bcl-2, and Fas/FasL signaling pathways and upregulating CGRP content.

Brain Ischemia ; Cerebral Infarction ; Humans ; Subarachnoid Hemorrhage ; Vasospasm, Intracranial/etiology*

Objective To investigate the effect of exosomes derived from Echinococcus multilocularis on macrophage polarization after treatment for different durations and concentrations. Methods A total of 60 BALB/c mice were used for modeling, among which 4 mice were selected to observe the growth of abdominal lesions on 7.0T MRI. The mice for modeling were dissected, and the protoscoleces was taken from the abdominal lesion and cultured in vitro ; ultracentrifugation was used to extract the exosomes from the supernatant, and transmission electron microscopy and Western blotting were used for the characterization of exosomes. The macrophages without exosome treatment were established as control group, and the macrophages co-cultured with different concentrations of exosomes derived from Echinococcus multilocularis were established as experimental group (10 μg/mL group and 50 μg/mL group) and were cultured for 48 and 72 hours. The morphological changes of macrophages were observed under a microscope, and flow cytometry and ELISA were used to observe polarization state. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results The results of 7.0T MRI showed the formation of diffuse lesions with different sizes in the abdominal cavity of mice, and the exosomes derived from Echinococcus multilocularis were approximately 100 nm in diameter and were cup-shaped or saucer-shaped, with the positive expression of the surface markers CD9, TSG101, and CD63. After co-culture, most of the cells in the experimental group were elongated with an irregular and polygonal shape. Flow cytometry showed that after 48 hours of co-culture, the positive rates of CD16/32, CD206, and CD369 in the control group were 99.53%±0.06%, 90.27%±0.21%, and 2.40%±0.20%, respectively; compared with the control group, except that the 10 μg/mL exosome group had a significant reduction in the positive rate of CD369 (0.80%±0.00%) ( P < 0.05), all the other groups had a significant increase in the positive rates of CD16/32, CD206, and CD369 (all P < 0.000 1); after 72 hours of co-culture, the positive rates of CD16/32, CD206, and CD369 in the control group were 99.67%±0.06%, 85.47%±0.55%, and 6.60%±0.20%, respectively, and compared with the control group, the experimental group had significant increases in the positive rates of CD16/32, CD206, and CD369 (all P < 0.05). ELISA showed that after 48 hours of co-culture, the levels of IL-6 and TNFα in the control group were 58.53±15.52 pg/mL and 320.70±5.30 pg/mL, respectively, and when the exosome concentration was 50 μg/mL, the level of IL-6 in the experimental group was 98.81±15.55 pg/mL, which was higher than that in the control group ( P < 0.05); after 72 hours of co-culture, the levels of IL-6 and TNFα in the control group were 76.22±9.68 pg/mL and 323.90±87.37 pg/mL, respectively, and when the exosome concentration was 10 μg/mL, the level of TNFα was 164.20±14.17 pg/mL, which was significantly lower than that in the control group ( P < 0.05); when the exosome concentration was 50 μg/mL, the level of IL-6 was 99.52±8.35 pg/mL, which was significantly higher than that in the control group ( P < 0.05). Conclusion Exosomes derived from Echinococcus multilocularis can regulate macrophage polarization and induce M2-like polarization of macrophages after co-culture at a concentration of 10 μg /mL for 72 hours, and further studies are needed to clarify the specific method.