[Role of alkaloid sinomenine in chronic rejection in the rat heart transplantation model].

OBJECTIVE: To evaluate the possible role of alkaloid sinomenine (SIN) on chronic rejection in rat heart transplantation model. METHODS: After a brief course of cyclosporine A (CsA), DA recipients of PVG hearts were treated with placebo, SIN, CsA, or a combination of both drugs. Grafts were analyzed morphometrically and by immuno-histochemistry. Expressions of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and endothelin 1 (ET-1) were assessed by reverse transcription-polymerase chain reaction. RESULTS: Cardiac grafts of SIN-treated rats showed a mild degree of vasculopathy compared with untreated rats or CsA-treated recipients. Degree of vasculopathy was significantly reduced in rats treated with combined SIN and CsA than rats receiving either drug alone. Treatment with SIN alone did not affect gene expressions of bFGF, VEGF, and ET-1 while expressions of bFGF, VEGF, and ET-1 were significantly reduced by combined treatment with SIN and CsA. CONCLUSION: These results demonstrated a potential value of SIN, in combination with low-dose CsA to attenuate the vasculopathy in this rat model of chronic cardiac allograft rejection.

Effect of 43 degrees treatment on expression of heat shock proteins 105, 70 and 60 in cultured monkey Sertoli cells / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To examine the possible effect of heat treatment on expression of heat shock proteins (Hsps) 105, 70, and 60 in primary monkey Sertoli cells and to evaluate the possible signal pathways.</p><p><b>METHODS</b>Western blot analysis, real-time polymerase chain reaction (PCR), and confocal immunohistochemistry were used to analyze mRNA and protein levels of the Hsps in response to 43 degrees treatment of Sertoli cells isolated from pubertal monkey testes.</p><p><b>RESULTS</b>Staining with Hoechst 33342 indicated Sertoli cells did not undergo apoptosis after heat treatment. Hsp105 was expressed in cytoplasm of untreated Sertoli cells. Both Hsp105 mRNA and protein levels were increased approximately 20-fold compared to those of the untreated controls at 12 h after heat treatment. Untreated Sertoli cells did not express Hsp70, but heat stress induced its expression in the cell cytoplasm. The time-course of changes in Hsp70 was similar to that of Hsp105. In contrast to Hsp105 and Hsp70, the change in Hsp60 expression was much less obvious. The protein level between 12 h and 48 h after heat treatment was only approximately 1.5-fold that of the untreated control. Extracellular regulated kinase (ERK) 1/2 inhibitor (U0126) or phosphoinositide kinase-3 (PI3K) inhibitor (LY294002) could partially block the response of Hsp105 and Hsp70 induced by heat treatment.</p><p><b>CONCLUSION</b>These results indicate that the heat-induced expression of the three types of Hsp in monkey Sertoli cells might be regulated by ERK and/or PI3K signal pathways, but the profile of their expression is different, suggesting that they might have different regulatory functions in Sertoli cells.</p>

[Effects of the alkaloid sinomenine on COX-2 activity and acute rejection in heart allografts: an experiment with rats].

OBJECTIVE: To evaluate the effects of the alkaloid sinomenine (SIN), a COX-2 inhibitor, on the acute rejection in heart allografts. METHODS: Forty Wistar rats received the allograft of the hearts of 40 SD rats into the peritoneum. Then the recipients were randomly divided into 2 groups: SIN group, injected with SIN within 24 hours after the operation; and control group, injected with normal saline. The survival time was observed. The heartbeat was examined every day. The Wistar rats were killed 3 and 5 days after the operation respectively and the left ventricular tissues were taken to undergo pathological examination to detect the acute rejection and cell apoptosis. Immunochemistry and Western blotting were used to detect the COX-2 protein, and RT-RCR was used to detect the COX-2 mRNA. The mean numbers of apoptotic cardiomyocytes were determined with the terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL) technique. RESULTS: The survival time of the SIN group was 12.5 +/- 2.6 days, significantly longer than that of the control group (6.8 +/- 0.5 days, P = 0.001). Examination 3 and 5 days after the treatment of SIN, the extents of inflammatory reaction, endovasculites, myocardial edema, and cardiomyocyte damage in the allografts of the SIN group were all significantly less, the mean numbers of apoptotic cardiomyocyte was significantly smaller compared with the control group (all P < 0.05). At day 5, the levels of COX-2 protein and mRNA of the SIN group were both significantly lower than those of the control group. CONCLUSION: Inhibition of COX-2 prolongs the allograft survival and reduces the myocardial damage and inflammation during acute cardiac allograft rejection.

Activation of extracellular signal-related kinases 1 and 2 in Sertoli cells in experimentally cryptorchid rhesus monkeys / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To assess the spatiotemporal changes in the expression of extracellular signal-regulated kinases 1 and 2 (ERK1/2), c-Jun N-terminal kinases (JNK) and p38 mitogen-activated protein kinases (MAPK) in response to heat stress in the cryptorchid testis, and to investigate a possible relation to Sertoli cell dedifferentiation.</p><p><b>METHODS</b>Immunohistochemistry and western blot were used to examine the expression and activation of ERK1/2, p38 and JNK in the cryptorchid testis at various stages after experimental cryptorchidism.</p><p><b>RESULTS</b>The abdominal temperature did not obviously change the total ERK1/2 expression but significantly activated phospho-ERK1/2 in the Sertoli cells of the cryptorchid testis. Heat stress increased total JNK expression in the Sertoli cells of the cryptorchid testis but did not activate phospho-JNK. Neither total p38 nor phospho-p38 was induced by heat stress in the Sertoli cells of the cryptorchid testis. Changes in the spatiotemporal expression of cytokeratin 18 (CK18), a marker of immature or undifferentiated Sertoli cells, were induced in the cryptorchid testis in a pattern similar to the activation of ERK1/2.</p><p><b>CONCLUSION</b>The activation of ERK1/2 in the testis may be related to dedifferentiation of Sertoli cells under heat stress induced by experimental cryptorchidism.</p>