Objective To study the effects of heat shock proteins (HSPs) in the course of hepatic ischemia reperfusion injury (HIRI), and analyze its mechanism. Methods The relationship between HSPs and HIRI was studied by reviewing literatures. Results HSPs was a kind of stress protein induced after cell was sitmulated by the stress. It could improve body′s tolerance to tough situation. Though hepatic ischemia reperfusion usually results in serious hepatic injury, at the same time it could induce can increase the production of HSPs that can protect liver from and lessen ischemia reperfusion injury. Conclusion HSPs can improve the tolerance to HIRI and lessen injury. In addition, HSPs is thought to be markers of HIRI, and can be used as a efficient indicator to test the level of hepatic injury and assess prognosis.
Objective To observe the expression of heat shock protein 70 (HSP70) in human liver after hepatic transplantation, and to study its correlation with the occurrence and progression of acute allograft rejection.Methods Fifteen biopsy specimen of allograft liver after transplantation were collected and divided into three groups according to their pathological changes: control group (no rejection), mild acute rejection group, and moderate/serious acute rejection group. The expressions of HSP70 in grafts were detected by using immunohistochemical method and imaging analysis. Results HSP70 was expressed in all 3 groups, and appeared mainly in hepatocellular cytoplasm. The immunohistochemical imaging analysis of HSP70 showed: integral optical density (IOD) which was 30.99±11.14 in the control group was lower than that in the mild acute rejection group (68.84±21.37) and that in the moderate/serious acute rejection group (71.82±19.99), P＜0.01； and the IOD in the moderate/serious acute rejection group was higher than that in the mild acute rejection group (P＜0.05). Conclusion HSP70 plays a role in cellular protection for allograft liver, and the continuously increasing expression of HSP70 in graft maybe closely relates to the occurrence and progression of acute allograft rejection.
Objective To analyze the protective effects of heat-shock response on the retinae of the rats after retinal ischemic reperfusion injury.Method Twenty Wistar rats (20 eyes) were divided into 4 groups: intracameral perfusion group (group P), intracameral perfusion after quercetin injection group (group P+Q), intracameral perfusion after heat shock group (group P+H), and in tracameral perfusion after quercetin injection and heat shock group (group P+Q+H ). According to the standard program established by International Society for Clinical Visual Electrophysiology, we recorded the results of the dark-adapted electroretinogram (D-ERG ),oscillatory potentials (OPs),and light-adapted ERG (L-ERG) of the rats with intraocular hypertension after induced by heat shock response. The expressions of HSP 70 of the rats in all groups were observed by Western blotting.Results The expression of HSP 70 of the rats in group P+H was the highest in all groups, but the expressions of HSP70 in group P+Q and P+Q+H were inhibited significantly. The amplitudes of a and b wave of ERG and O2 wave of OPs decreased, and the delitescence of them were delayed significantly in rats after intracameral perfusion. The amplitude of b wave of D-ERG and O2 wave of OPs in group P+H were higher than which in group P. Zero hour after perfusion, the amplitudes of all waves in group P+H increased significantly (Plt;0.05). Twenty-four hours after perfusion, the retinal functional resumption of the rats in group P+H was better than which in group P. In group P+Q and P+Q+H, the delitescences of all waves of ERG and O2 wave of OPs were the longest and the amplitudes were the lowest, and some waves even disappeared.Conclusions The heat-shock response may improve the recovery ability of the retinal cells after injury of ischemic reperfusion.(Chin J Ocul Fundus Dis，2003，19：117-120)
ObjectiveTo investigate the expression of heat shock proteins 90α(HSP90α) in human hepatocellular carcinoma and the relationship between its expression and biologic behavior of tumor and prognosis. MethodsUsing the immunohistochemical SP method, HSP90α expression was detected in liver tissue from 10 normal individuals, 40 patients with hepatocellular carcinoma(HCC) and adjacent noncancerous liver tissues. ResultsThe positive expression rate of HSP90α was 10.0%,52.5%,72.5% in normal liver tissues,adjacent noncancerous liver tissues,hepatocellular carcinous tissues respectively. A significantly higher distribution of HSP90α positive expression in HCC tissues compared with adjacent noncancerous liver tissues and normal liver tissues was obtained （P＜0.05）. The positive expression of HSP90α in HCC was correlated with clinical stage, tumor differentiation, serosal condition and lymph node metastasis （P＜0.05）, but not to tumor number （P＞0.05）. It was also correlated with prognosis of HCC. The mean tumorfree survival of patients with HSP90α negative expression was 38.6 months while that of HSP90α positive expression was 25.5 months （P＜0.05）. ConclusionHSP90α is overexpressed in human hepatocellular carcinoma. HSP90α could be used as an indicator to judge the clinical stage, tumor differentiation, serosal condition, lymph node metastasis and prognosis of HCC.
Objective To study the expression of heat shock protein 47 (HSP47) and its correlation to collagen deposition in pathological scar tissues. Methods The tissues of normal skin(10 cases), hypertrophic scar(19 cases), and keloid(16 cases) were obtained. The expression ofHSP47 was detected by immunohistochemistry method. The collagen fiber content was detected by Sirius red staining and polarization microscopy method. Results Compared with normal skin tissues(Mean IOD 13 050.17±4 789.41), the expression of HSP47 in hypertrophic scar(Mean IOD -521 159.50±272994.13) and keloid tissues(Mean IOD 407 440.30±295 780.63) was significantly high(Plt;0.01). And there was a direct correlation between the expression of HSP47 and the total collagen fiber content（r＝0.386，Plt;0.05）. Conclusion The HSP47 is highly expressed in pathological scartissues and it may play an important role in the collagen deposition of pathological scar tissues.
Objective To investigate the role and mechanism of heat shock protein 60 (HSP60) in induction of murine skin allograft tolerance. Methods At the age of 8-12 weeks, inbred female BALB/C (H-2d) mice (n=45) and CBA/N (H-2k)mice (n=15) were used as transplantation donors and C57BL/6 (H-2b) mice (n=60) as recipients. Recipients C57BL/6 (H-2b) mice were randomized into 4 groups (n=15). In group A, 1 cm × 1 cm Wolfe-Krause skin graft was excised from the back of BALB/C (H-2d) mice and hypoderma was scraped off aseptically, and then transplanted to the back of C57BL/6 (H-2b)mice. The method of skin transplantation in the other 3 groups was the same as to group A. In group B, C57BL/6 (H-2b) mice were treated with imcompleted Freund’s adjuvant (IFA) administration into the back 2 weeks before transplantation of BALB/C (H-2d) mice skin. In group C, C57BL/6 (H-2b) mice were administered HSP60 emulsified in IFA into the back 2 weeks before transplantation of BALB/C (H-2d) mice skin. In group D, C57BL/6 (H-2b) mice were treated with HSP60 emulsified in IFA into the back and followed by skin transplantation of CBA/N (H-2k) mice 2 weeks later. The delayed type hypersensitivity was determined 7 days after transplantation. One-way mixed lymphocyte reaction, the concentration of cytokines in the mixed lymphocyte reaction culture supernatant was determined 7 days and 25 days after transplantation. The survival time of skin allograft was observed. Results The survival time of skin allograft in groups A, B, C and D was 12.4 ± 0.5, 11.6 ± 0.8, 29.3 ± 2.6 and 27.6 ± 2.1 days, respectively. There was significant difference between groups A, B and groups C, D (P﹤0.05), while there was no significant difference between group A and group B as well as between group C and group D (P gt; 0.05). The counts of per minute impulse (cpm) of mixed lymphocyte reaction 7 days after transplantation in groups A, B, C and D was 12 836 ± 1 357, 11 876 ±1 265, 6 581 ± 573 and 6 843 ± 612, respectively. There was significant difference between groups A, B and group C and group D (P lt; 0.05), while there was no significant difference between group A and group B as well as between group C and group D (P gt; 0.05). The cpm of mixed lymphocyte reaction at 25 days after transplantation in group A, B, C and D was 13 286 ±1 498, 12 960 ± 1 376, 11 936 ± 1 265 and 12 374 ± 1269, respectively. There was no significant difference among 4 groups (P gt;0.05).The concentration of IL-10 in the mixed lymphocyte reaction culture supernatant in groups C, D were higher than that in groups A, B, and IL-2 and IFN-γ were lower than that in groups A, B 7 days after transplantation (P lt; 0.05), while there was no significant difference between group A and group B as well as between group C and group D (P gt; 0.05). There was no significant difference in cytokines among the 4 groups 25 days after transplantation (P gt; 0.05). The delayed type hypersensitivity in groups A, B, C and D 7 days after transplantation was 0.84 ± 0.09, 0.81 ± 0.07, 0.43 ± 0.05 and 0.46 ± 0.03 mm, respectively. There was significant differences between groups A, B and groups C, D (P lt; 0.05). While there was no significant difference between group A and group B as well as between group C and group D (P gt; 0.05). Conclusion HSP60 may play a role in induction and maintenance of murine skin allograft tolerance.
Objective To study the relationship between the expression ratio of heat shock protein (HSP) 70 to C-fos in organs outside the brain after brain concussion and the time of injury in rats, in order to provide a new visual angle for determining injury time of brain concussion. Methods The model of brain concussion was established through free falling method. Then the rats were executed at 30 minutes, 1 hour, and 3, 6, 12, 24, 48, 96, 168, 240, 336 hours after injury. Immunohistochemistry staining of C-fos and HSP70 were used in the materials from the main organs including heart, liver, spleen, lung and kidney. All related experiment results were studied by using a microscope with image analytical system and homologous statistics. Results From 30 minutes to 6 hours after injury, the proportion of HSP70 immuno-positive cells increased slowly, while the proportion of C-fos immuno-positive cells increased rapidly, and the ratio of HSP70/C-fos positive cells was on the decline. From 6 to 12 hours after injury, the proportion of HSP70 immuno-positive cells rose continuously, while the proportion of C-fos immuno-positive cells started to decrease, and the HSP70/C-fos ratio showed a rising tendency. From 12 to 336 hours after injury, the proportion of HSP70 immuno-positive cells decreased slowly, while the proportion of C-fos immuno-positive cells decreased rapidly, and the HSP70/C-fos ratio was still on the rise. Conclusions The proportion of positive cells and ratio of the two markers in the main organs including heart, liver, spleen, lung and kidney are similar to those in the brain of rats after brain concussion. Observing the proportion of positive cells of the two markers together with their ratio in the main organs outside the brain may provide a reference for the determination of injury time after brain concussion.
Objective To detect the expression of heat shock protein 47 mRNA in pathological scar tissue by using real-time fluorescent quantitative reversetranscription-polymerase chain reaction (RT-PCR). Methods The tissues of normal skin(n=6), hypertrophic scar(n=6) and keloid(n=6) were adopted, which were diagnosised by Pathology Department. Based on fluorescent TaqMan methodology, the real-time fluorescent quantitative RT-PCR were adopted to detect the expression ofheat shock protein 47 mRNA. Results Compared with normal skin tissue（0.019±0.021）×105, the expressions of heat shock protein47 cDNA of hypertrophic scar tissue（1.233±1.039）×105 and keloid tissue（1.222±0.707）×105 were higher， being significant differences（Plt;0.05）. Conclusion A fluorescent quantitative method was successfully applied to detecting the expression of heat shock protein 47 mRNA. Heat shock protein 47 may play an important role in promoting the formation of pathological scar tissue.
Objective To study hyperthermia induced apoptosis and the effect of aspirin on hyperthermia induced apoptosis in retinoblastoma cells. Methods Retinoblastoma cells (Y79) were divided into two groups:hyperthermia groups,hyperthermia+aspirin (0.18～18mu;g/ml) groups.Heat shock condition:44℃,heat shock time:10,20,30, and 40 minutes respectively.The following events were studied after heat shock by using FAC Scan: ①cell apoptosis; ②heat shock protein 70 (HSP70) expression；③bcl-2 expression. Results Apoptosis was induced by the treatment of hyperthermia (44℃) in Y79 cells in a heat dose dependent fashion.Longer time heating (44℃，40 minutes) induced necrosis rather than apoptosis.Aspirin could rescue Y79 cells from hyperthermia induced apoptosis in a dose dependent manner．HSP70 was induced in Y79 cells after heat shock,it was further enhanced by the treatment of aspirin(＞1.8mu;g/ml).Heat shock itself showed no effect on bcl-2 expression in Y79 cells,aspirin,on the other hand,could enhance bcl-2 expression in a modest level in heat treated Y79 cells. Conclusions Hyperthermia may induce apoptosis in Y79 cells which can be protected by use of aspirin．The enhancement of HSP70 and bcl-2 expression in Y79 cells by the treatment of aspirin in heating condition may be responsible for the protective function. (Chin J Ocul Fundus Dis, 1999, 15: 143-145)
By using biochemical assessment technique and histological examination,a comparative study of the cutaneous tissues in 16 patients with lymphedema of the lower extremity before and after the heating and bandage therapy, and it was noted thatthe heating and bandage therapy might:(1) the content of hydroxyproline in the affected skin would be decreased; (2) the thickness of skin was decreased and the water content was reduced; (3) the microcirculation of local tissues was enhanced, and (4) the activity of the macrophages was increased. In conjunction with the criteria of clinical observation, the action mechanism of heating and bandage therapy might be as follows: (1) improve the local microcirculation and enhance the resorption of tissue fluid and the protein, and (2) increase the activity of the macrophages, and minimize the extent of fibrosis of the affected tissues.