Objective To investigate the influence of Chaihu Longgu Muli Tang(Chinese Thorowax Root, Bone Fossil of Big Mammals and Oyster Shell Decoction, CLMT) on the mobilization of bone marrow c-kit+stem cells in rats after myocardial infarction(MI). Methods Male SD rats were randomly divided into sham-operation group,model group and CLMT group,and each group was divided again into 3-d sub group,7-d subgroup and 14-d subgroup. The CLMT group was intragastrically given CLMT, and other groups were orally given distilled water. The heart function was reviewed by using echocardiogram, changes of myocardial pathology were reviewed after HE staining,and severity of myocardial fibrosis was observed by after Masson staining. The number of c-kit+positive cells in bone marrow and peripheral blood was detected by using flow cytometry (FCM), and number of c-kit+positive cells in MI border zone was detected by using immunohistochemistry technique. Results The heart function declined gradually along with MI time extension in model group, and difference in heart function had statistical significance compared with sham-operation group at all time points (P <0.05). The heart function trended to stable in CLMT group and was better than that in model group, and difference in LVEF and LVFS had statistical significance compared with model group at time points of 7 d and 14 d(P<0.05). Pathological staining showed that myocardial cells were in alignment without collagen fibers in sham-operation group,necrosis and fibrosis of myocardial cells in model group that aggravated gradually along with MI time extension. The severity of myocardial necrosis and fibrosis was milder in CLMT group than that in model group. The number of c-kit+positive cells in bone marrow and peripheral blood reached peak in model group and CLMT group 7 d after MI, and difference number of c-kit+positive cells had statistical significance compared with sham-operation group,and was higher in CLMT group than that in model group. The difference in number of c-kit+positive cells had statistical significance between 7-d subgroup and 14-d subgroup (P <0.05). The number of c-kit+positive cells in MI border zone was higher in model group than that in sham-operation group,and difference in number of c-kit+positive cells had statistical significance between 7-d subgroup and 14-d subgroup (P<0.05). The number of c-kit+positive cells in MI border zone was higher in CLMT group than that in model group, and difference in number of c-kit+positive cells MI border zone had statistical significance between 7-d subgroup and 14-d subgroup (P<0.05). Conclusion CLMT can improve the mobilization of bone marrow c-kit+stem cells after MI and heart function.

Objective:To evaluate the relationship between milk fat globular epidermal growth factor Ⅷ (MFG-E8)-mediated efferocytosis and sepsis-associated encephalopathy (SAE) in mice.Methods:Forty clean-grade healthy male C57BL/6 mice, aged 2-3 months, weighing 18-22 g, were divided into 4 groups ( n=10 each) using a random number table method: sham operation group (group Sham), cecal ligation and perforation (CLP) group (group CLP), sham operation+ phosphate buffer solution (PBS) group (group Sham+ PBS) and CLP+ recombinant mouse MFG-E8 group (group CLP+ rmMFG-E8). SAE was induced by CLP in anesthetized mice.PBS 1 μl and rmMFG-E8 1 μg were injected into the lateral cerebral ventricle in Sham+ PBS and CLP+ rmMFG-E8 groups after operation for 5 consecutive days.Novel object recognition test was performed at 6 days after operation, and the contextual fear conditioning test was performed at 7 and 8 days after operation.The percentage of time spent exploring a novel object, discrimination index and percentage of freezing time induced by condition were calculated.The animals were sacrificed after the end of behavioral tests, and the hippocampus was extracted for determination of the expression of MFG-E8 and GTP-Rac1 (by Western blot), the mRNA expression levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α and IL-1β (using real-time reverse transcription-polymerase chain reaction) and the apoptosis rate in hippocampus (by TUNEL). Results:Compared with group Sham, the percentage of time spent exploring a novel object, discrimination index and percentage of freezing time were significantly decreased, hippocamcal MFG-E8 expression was down-regulated, GTP-Rac1 expression was up-regulated, mRNA expression of IL-6, TNF-α and IL-1β was up-regulated, and apoptosis rate was increased in group CLP ( P<0.05). Compared with group CLP, the percentage of time spent exploring a novel object and discrimination index were significantly increased, freezing time was prolonged, hippocamcal MFG-E8 and GTP-Rac1 expression was up-regulated, mRNA expression of IL-6, TNF-α and IL-1β was down-regulated, and apoptosis rate was decreased in group CLP+ rmMFG-E8 ( P<0.05). Conclusion:The reduction of hippocampal MFG-E8-mediated efferocytosis may be involved in the pathophysiological mechanism of SAE in mice.