二氢卟吩e6在艾氏荷瘤小鼠体内的分布

目的研究二氢卟吩e6(Ce6)在移植瘤小鼠体内吸收、分布及代谢的动态变化,以期为声动力疗法处理不同部位肿瘤的时间点提供科学依据。方法艾氏移植瘤小鼠尾静脉注射Ce6后,利用荧光分光光度法和小动物活体成像技术测定Ce6在小鼠不同组织的富集分布变化规律。结果小鼠尾静脉给药后,Ce6迅速分布于全身各组织,在2 h内,各组织药物浓度均达到峰值,其中以肝含量最高。随后各组织中药物浓度均开始下降,以肝中清除速度最快。肿瘤组织中的Ce6含量在注射后不断上升,2 h时达到最高,随后开始下降,2～10 h代谢比较缓慢,24 h时浓度降至最低,但仍高于其他组织。结论 Ce6在艾氏移植瘤中具有肿瘤组织选择性好、潴留时间长并可迅速从体内排出等优点,有着很好的临床应用前景,同时提出了不同组织类型不同部位的肿瘤应根据各自适当的时间点进行处理。     

Physicochemical Characterization and Study of in vitro Interactions of pH-sensitive Liposomes with the Complement System

Complement activation is an important step in the acceleration of liposome clearance. The anaphylatoxins released following complement activation may motivate a wide variety of physiologic changes. We performed physicochemical characterization and in vitro studies of the interaction of complement system with both noncirculating and long-circulating pH-sensitive and nonpH-sensitive liposomes. The liposomes were characterized by diameter, zeta potential, and atomic force microscopy (AFM). The study of liposome interactions with complement system was conducted using hemolytic assay in rat serum. All liposomes presented a similar mean diameter (between 99.8 and 124.3 nm). The zeta potential was negative in all liposome preparations, except in liposomes modified with aminopoly (ethyleneglycol) 2000-distearoylphosphatidylethanolamine (aPEG₂₀₀₀-DSPE), which presented positive zeta potential. Atomic force microscopy images showed that non–long-circulating pH-sensitive liposomes are prone to vesicles aggregation. Non–pH-sensitive liposomes complement system activates, while pH-sensitive liposomes showed to be poor complement activators in rat serum.     

Tissue distribution and antileishmanial activity of liposomised Amphotericin-B in Balb/c mice§

Antileishmanial activity and organ distribution of the antifungal drug Amphotericin-B in free and liposomised form have been studied in Balb/c mice infected withLeishmania donovani. Results indicate that Amphotericin-B in the liposomised form is significantly more active than the free form. This increase in the activity is perhaps related to the reduced drug toxicity rather than the altered drug distribution at the site of infection. CDRI communication No. 4789     

Tissue distribution and hormonal regulation of the breast cancer resistance protein (Bcrp/Abcg2) in rats and mice

Breast cancer resistance protein (Bcrp/Abcg2) is a member of the ABC transporter family. The purpose of this study was to quantify Bcrp mRNA in rat and mouse tissues, and to determine whether there are gender differences in Bcrp mRNA expression. Rat Bcrp mRNA levels were high in intestine and male kidney, and intermediate in testes. Mouse Bcrp expression was highest in kidney, followed by liver, ileum, and testes. Male-predominant expression of Bcrp was observed in rat kidney and mouse liver. Furthermore, gonadectomy and hypophysectomy experiments were conducted to determine whether sex steroids and/or growth hormone are responsible for Bcrp gender-divergent expression patterns. Male-predominant expression of Bcrp in rat kidney appears to be due to the suppressive effect of estradiol, and male-predominant expression of Bcrp in mouse liver appears to be due to the inductive effect of testosterone.     

Mechanisms mediating metabolic abnormalities in the livers of Ehrlich ascites tumor-bearing mice

Previously we reported that intermittent intraperitoneal administration of ornithine decarboxylase-inducing factor (ODC factor), interleukin 1alpha (IL-1alpha), and tumor-necrosis factor-alpha (TNF-alpha) to normal mice induced biological changes in the hosts which included changes in the pattern of expression of pyruvate kinase (PK) isozymes in the liver and hypertrophy of the spleen. In the study reported here, we investigated the chronic and combined effects of these factors on hepatic enzymes using alzet microosmotic pumps implanted in the subcutis of the backs or abdominal cavities of mice. Continuous administration of ODC factor and recombinant human IL-1alpha (rhIL-1alpha) reduced the activity of L-type PK, which is a glycolysis-related enzyme in the liver, and induced the activity of M2-type PK, a known marker of liver dedifferentiation. Serine dehydratase (SDH) and tyrosine aminotransferase (TAT), enzymes associated with amino acid metabolism, were not significantly influenced at the examined concentration. The simultaneous continuous infusion of ODC factor and rhIL-1alpha or rhTNF-alpha caused alterations in the patterns of expression of PK isozyme activity profiles and reduced overall PK activity. SDH and TAT activities were also significantly induced. Moreover, mice treated with these combined factors displayed many other metabolic changes normally associated with cancer cachexia. These findings suggest that the tumor-derived ODC factor and cytokines such as IL-1alpha and TNF-alpha might function synergistically in the metabolic perturbations observed in Ehrlich ascites tumor bearers.     

Tissue specification and intracellular distribution of actin isoforms inVicia faba L.

Summary Two dimensional gel electrophoresis of total cell protein extracts from not expanded, and primary leaves, petioles, and roots ofVicia faba resulted in four actin isoforms at 43 kDa with pI values from 5.9 to 6.05. In contrast to root extracts, in all leaf extracts an additional immunoreactive polypeptide with a molecular mass of 51 kDa and pI 5.75 was detected. This polypeptide was present in high amounts in protein extracts of purified chloroplasts, whereas no actin isoform at 43 kDa could be demonstrated. Compared to the tissue extracts, two actin isoforms at 43 kDa with pI values of 5.9 and 6.0 were enriched, when purified plasma membranes and the membranous fraction of vacuoles were analysed. In contrast, the soluble protein fraction of the plasma membrane preparation contained only two isoactins with pI values of 5.95 and 6.05 and a molecular mass of 43 kDa. These results indicate, that the four actin isoforms at 43 kDa detected in all examined tissues ofV. faba fulfill different functions at specific intracellular compartments, for example, the anchorage of actin microfilaments to membranes.Abbreviations BSA bovine serum albumin - BCIP 5-bromo-4-chloro-3-indolyl phosphate - DDM n-decyl -D-maltopyranoside - EDTA ethylenediamine-tetraacetic acid - HG n-hexyl -D-glucopyranoside - IEF isoelectrical focusing - MES morpholinoethanesulfonic acid - 2-ME 2 mercaptoethanol - NBT nitro blue tetrazolium - pCMB p-chloromercuribenzoic acid - PVP polyvinylpyrrolidone - Tris tris (hydroxymethyl) aminomethane     

Pharmacokinetics and tissue distribution of novel traditional Chinese medicine-platinum anticancer agents in rats

The pharmacokinetics and tissue distribution profiles of a novel series of traditional Chinese medicine-platinum (TCM-Pt) compounds [Pt(C(8)H(8)O(5))(NH(2)R)(2)]: 1 (where R=H), 3 (R=CH(3)) and 5 (R=C(6)H(10)), were studied in Sprague-Dawley rats following a single bolus intravenous (i.v.) injection. Platinum concentrations in total plasma, plasma ultrafiltrate, urine and tissues were measured by flameless atomic absorption spectroscopy. Pharmacokinetic studies showed that plasma concentrations of total and free platinum for the novel TCM-Pt compounds as well as cisplatin and carboplatin declined in a biexponential manner with a short distribution half-life (t(1/2alpha): 0.12-0.34h). Compared with cisplatin, the novel TCM-Pt compounds had a longer elimination half-life (t(1/2beta)), larger dose normalized area under the curve (AUC/D), larger volume of distribution at steady-state (V(ss)), slower clearance (CL) of free platinum and higher percentage of cumulative urinary excretion (CUE), which can be attributed to their lower chemical reactivities. In tissues, the highest Pt concentrations were found in the kidney, followed by the liver and the lowest in the heart; no Pt was detected in the brain. Twenty-four hours after drug administration, platinum concentrations in tissues were significantly lower for the novel TCM-Pt compounds. These findings suggest that the novel compounds might afford higher clinical efficacy and reduced systemic side effects, when compared with cisplatin.     

Effects of alkyl glycosides incorporated into liposomes prepared from synthetic amphiphiles on their tissue distribution in Ehrlich solid tumor-bearing mice.

A study of the effects of alkyl glycosides incorporated into synthetic liposomes with respect to their stability, their in vivo distribution in Ehrlich solid tumor-bearing mice and their in vitro interaction with liver cells was undertaken. The synthetic liposomes were prepared from N,N-didodecyl-N alpha-[6-(trimethylammonio)hexanoyl]-L-alaninamide bromide (N+C5Ala2C12) and labeled with 99mTc. n-Dodecyl glucoside (DG) and n-dodecyl sucrose (DS) were used as alkyl glycosides. The stability was hardly changed by incorporation of alkyl glycosides into the liposomes in saline and serum. The uptake of DG- and DS-modified N+C5Ala2C12 liposomes decreased in liver and spleen compared with that of unmodified N+C5Ala2C12 liposomes, resulting in an increase in blood and other tissues such as tumor, duodenum and kidney, where the DS-modified N+C5Ala2C12 liposomes had a marked tendency. It was observed with electron micrographs that the size of N+C5Ala2C12 liposomes became small by incorporation of alkyl glycoside. The smaller N+C5Ala2C12 liposomes were found to result in the lower uptake in liver. The interaction of the liposomes with liver cells in vitro indicated that both DG- and DS-modified liposomes had a low affinity for liver cells compared with the unmodified liposomes and the extent of interaction of the DS-modified liposomes was weaker than that of the DG-modified liposomes.     

Antigenotoxic and anticytotoxic effect of camel milk in mice treated with cisplatin

Camel milk (CM) has good nutritive value, in addition to its antigenotoxic and anticytotoxic effects. Therefore the aim of this investigation was to evaluate the capacity of CM to inhibit the micronucleated polychromatic erythrocytes (MnPCEs) in the bone marrow and improve the mitotic activity produced by cisplatin. Cisplatin is one of the most widely used antineoplastic drugs in the treatment of cancer. The 70 adult male Swiss albino mice were divided into seven groups:

• Gr. I: treated with distilled water and considered as a control group.
• Gr. II: treated with camel milk (33 ml/kg, b.w).
• Gr. III: treated previously with cisplatin (0.5 mg/kg, b.w).
• Gr. IV: treated with camel milk and followed after 2 h. with cisplatin (33 ml/kg → 0.5 mg/kg, b.w).
• Gr. V: treated with camel milk and cisplatin at the same time (33 ml/kg + 0.5 mg/kg, b.w).
• Gr. VI: treated with an acute single dose of cisplatin (2.5 mg/kg, b.w).
• Gr. VII: treated with camel milk prior and followed with an acute single dose of cisplatin (33 ml/kg → 2.5 mg /kg, b.w). The animals were sacrificed 24 h after cisplatin injection. The pretreatment with CM dose caused a significant decrease (P < 0.001) in the frequency of MnPCEs and increase (P < 0.001) in the mitotic index (MI) induced by cisplatin when compared with the groups treated with cisplatin alone. The possible explanation for the antigenotoxic and anticytotoxic effects observed in the pretreatment with CM is ascribed to its contents. In conclusion, from the findings we suggest that this milk has some antioxidant effect, and the antigenotoxic mechanism of this milk needs to be explored further before their use during cisplatin chemotherapy.

     

黄瓜香联合顺铂治疗小鼠H22肝癌的研究

目的 观察黄瓜香联合顺铂对小鼠H22肝癌移植瘤生长的抑制作用.方法 将40只接种H22肝癌细胞的昆明小鼠随机分为4组,其腹腔分别注射生理盐水(对照组);黄瓜香组;顺铂组;黄瓜香+顺铂组(联合治疗组),观察小鼠的毒副反应及生存质量.实验19 d后,处死全部小鼠,剥离皮下肿瘤,称小鼠肿瘤重量,计算抑瘤率.结果 黄瓜香组的H22肝癌平均瘤重为(1.26 ±0.19)g,明显低于对照组的(1.89 ±0.56)g(P ＜0.01).联合治疗组的平均瘤重为(0.57 ±0.42)g,均明显低于黄瓜香组(P＜0.01)和顺铂组(P＜0.01);其抑瘤率达69.8％,明显高于黄瓜香组(x2=16.9875,P＜0.01)和顺铂组(x2=5.0602,P＜0.05).联合治疗组小鼠的毒副反应明显低于顺铂组,生存质量好于顺铂组;黄瓜香组与联合治疗组都能调节肠道菌群,扶植肠道中有益菌(乳酸杆菌和双歧杆菌)生长,抑制大肠埃希菌生长,与对照组比较差异具有统计学意义(P＜0.01).结论 黄瓜香与顺铂合用对小鼠H22肝癌移植瘤的生长具有协同抑制作用,能降低顺铂毒副反应,提高小鼠的生存质量.     

Tissue distribution of isoflavones in ewes after consumption of red clover silage

When discovered in the 50’s, isoflavones were suspected to provoke infertility syndrome in sheep grazing on clover. Many others effects of these phytoestrogens have been documented afterwards. To determine the distribution of isoflavone metabolites in ewe tissues and look for a link with their physiological impact, two ewes were fed a diet containing 50% red clover silage (variety Pawera) for one month with a daily intake of 157.6 mg/kg bw of total isoflavones. Only aglycones were fed due to the fermentation stage of the silage. At the sacrifice, isoflavone metabolites and aglycones were analyzed in blood, liver, kidney, lung, heart, muscle, ovaries, uterus, mammary glands, suprarenal glands, thymus, aorta, thyroid, pituitary gland, cerebellum, olfactory lobes, and brain hemispheres using HPLC-Coularray and LC-MS-MS. The major compounds recovered in tissues were equol and daidzein, present as glucuronides. Kidney concentrations were 10-fold higher than in other tissues. Penetration in brain was very limited. Reproductive organs contained higher concentrations of isoflavones than heart, muscle, or thymus. Distribution of isoflavones in ewe tissues is unequal and may reflect specific impact in some target tissues.     

Tissue distribution of humanized anti-human Fas monoclonal antibody (R-125224) based on fas antigen-antibody reaction in collagen-induced arthritis monkeys

R-125224 is a novel humanized anti-human Fas monoclonal antibody prepared from HFE7A, which is a monoclonal mouse IgG anti-Fas antibody, by grafting the mouse complementarity-determining regions to human IgG, presently being developed as a drug for treatment of rheumatoid arthritis. In the present study, we investigated the tissue distribution of radioactivity in cynomolgus monkeys with collagen-induced arthritis at the arm joint (CIA monkeys) after intravenous administration of (125)I-labeled R-125224 ((125)I-R-125224). At 168 h after administration, we observed a high radioactivity in the bone marrow, thymus, lungs, liver, adrenals, spleen, ovaries, axillary lymph node and mesenteric lymph node compared to the radioactivity in the plasma. These tissues and organs in human are reported to express Fas antigen, strongly suggesting a specific binding of (125)I-R-125224 to Fas antigen in cynomolgus monkeys. Semi-micro autoradioluminograms of arm joint showed that radioactivity is detected in pharmacological site, such as the bone marrow and articular cavity at 168 h. The kinetics in binding of R-125224 to activated monkey lymphocytes and hepatocytes was also investigated. K(d) values of activated lymphocytes and hepatocytes were 1.51+/-0.08 and 0.60+/-0.11 nM, respectively, which were similar to those values in human lymphocytes and hepatocytes, demonstrating that R-125224 cross-reacts with the monkey Fas antigen.     

Biochemical and morphological evaluation of the effects of propolis on cisplatin induced kidney damage in rats

Cisplatin (CP) is a chemotherapeutic agent used to treat various types of cancer; nephrotoxicity is the most common adverse effect of the drug. We investigated the protective effects of propolis against CP induced kidney injury. Thirty-six male rats were divided into six equal groups: untreated control group, 50 mg/kg/day propolis group, 100 mg/kg/day propolis group, single-dose 7 mg/kg CP group, 7 mg/kg CP + 50 mg/kg/day propolis and 7 mg/kg CP + 100 mg/kg propolis. Rats were sacrificed after 14 days and kidneys were removed for histopathological and biochemical analyses. We used hematoxylin & eosin and periodic acid-Schiff staining to evaluate kidney histopathology and we used the TUNEL technique to assess apoptosis. We also measured total oxidant status (TOS), total antioxidant status (TAS), oxidative stress index (OSI), ischemia-modified albumin (IMA) and malondialdehyde (MDA) levels in tissue and blood specimens. Normal morphology was observed in the control, 50 mg/kg/day propolis and 100 mg/kg/day propolis groups by light microscopy. Degeneration of tubule cells, edema and tubule dilation were increased in the CP group compared to the control group. Degeneration of tubule cells and dilation of Bowman’s spaces were decreased in the CP + 50 mg/kg/day propolis and CP + 100 mg/kg/day propolis groups compared to the CP group. Tubule dilation decreased significantly in the CP + 100 mg/kg propolis group compared to the CP group. Also, the 7 mg/kg CP group exhibited altered proximal tubule epithelial cells, loss of brush border and thickening of the parietal layer of Bowman’s capsule in glomeruli and basal laminae of tubules. A normal brush border was observed in the CP + 50 mg/kg/day propolis and CP + 100 mg/kg/day groups. Serum OSI and MDA levels were increased in the CP group compared to the control group. Serum MDA levels decreased significantly in the CP + 50 mg/kg/day propolis and 100 mg/kg CP + propolis groups compared to the CP group. CP caused significant damage to kidney tissue; propolis exhibited dose-dependent prevention of tissue damage.     

Distribution of trace elements in normal and tumor-bearing mice using the multitracer technique

A multitracer solution obtained from the nuclear reaction of selenium with 25-MeV/nucleon ⁴⁰Ar ions was orally administered to normal and tumor-bearing Balb/c male mice. After 96 h, the mice were sacrificed and the elemental distribution was determined in various tissues, organs, and blood. The uptake of Na, Rb, Ga, Sc, V, Cr, Mn, Co, Fe, Zn, Y, Zr, Tc, Ru, Ag, and In in normal and, except for zinc, in tumor-bearing mice was simultaneously detected. Most elements were distributed in about the same manner in the skin and liver of animals in both groups. The distribution of Rb, Ga, V, Cr, Tc, and In showed little or no significant differences between the two study groups. The distribution of Na, Mn, Fe, Ag, Sc, and Co showed significant differences between normal and tumor-bearing mice. In the blood, spleen, and kidney of the normal mice, there was good absorption of Na, Mn, Fe, Ag, Co, and Zn. In the heart, these elements were well absorbed, except for Na and Mn.     

Liposomal hamycin in the control of experimental aspergillosis in mice: relative toxicity, therapeutic efficacy and tissue distribution of free and liposomal hamycin.

Therapeutic efficacy of liposomal Hamycin has been evaluated in an animal model system for aspergillosis in Balb/c mice. Hamycin was intercalated into soya phosphatidyl choline (SPC), SPC: choline (1:1, vol./vol.) and DMPC liposomes. A single dose of either 0.1 mg/kg, 0.25 mg/kg or 0.5 mg/kg of liposomal Hamycin and 0.1 mg/kg of free Hamycin was injected (i.v.) into animals infected with Aspergillus fumigatus. An increase in the survival rate of animals along with decrease in fungal count in various organs was observed with liposomal administration. Incorporation of cholesterol into liposomes decreased the in vivo toxicity of Hamycin in a dose dependent manner. However, antifungal activity both in the presence and absence of cholesterol showed marked variation as compared to that of non-aromatic polyenes, e.g. amphotericin B. Analysis of Hamycin distribution by HPLC in various tissues revealed higher blood concentration of this drug, when given in free form, compared to its liposomised form. These studies suggest that liposomal Hamycin is more effective than free Hamycin in controlling the experimental Aspergillosis.     

Tissue distribution of bupivacaine enantiomers in sheep

rac-Bupivacaine HCl was infused intravenously to constant arterial blood drug concentrations in sheep using a regimen of 4 mg/min for 15 min followed by 1 mg/min to 24 h. At 24 h, arterial blood was sampled, the animal was killed with a bolus of KCl solution, then rapidly dissected and samples were obtained from heart, brain, lung, kidney, liver, muscle, fat, gut, and rumen. Tissue:blood distribution coefficients for (+)-(R)-bupivacaine exceeded those of (?)-(S)-bupivacaine (P < 0.05) for heart, brain, lung, fat, gut, and rumen by an overall mean of 43%. Blood:plasma distribution coefficients of (?)-(S)-bupivacaine exceeded those of (+)-(R)-bupivacaine by a mean of 29% and this offset the tissue:blood distribution coefficients so that the previously significant enantioselective differences disappeared. It is concluded that although enantioselectivity of bupivacame distribution is shown by the measured tissue:blood distribution coefficients, it is not shown when tissue:plasma water distribution coefficients are calculated, suggesting that there is no intrinsic difference between the bupivacaine enantiomers in tissue affinity. Sheep given fatal intravenous bolus doses of rac-bupivacaine had significantly greater concentrations of (+)-(R)-bupivacaine than (?)-(S)-bupivacaine in brain (P = 0.028) and ventricle (P = 0.036); these could augment the greater myocardial toxicity of this enantiomer found in vitro. © 1993 Wiley-Liss, Inc.     

Tissue distribution and clearance of soluble murine TNF receptors in mice

In this study, clearance of sTNFR was investigated. The data show that bilateral nephrectomy results in an increase of the levels of both sTNFR after which a new steady state situation develops, suggesting that other organs, apart from the kidneys, are involved in clearing of sTNFR. Bilateral nephrectomy also leads to an increase in circulating TNF. This TNF was detected by ELISA and appeared to be not biologically active. To investigate whether the endotoxin induced increase in sTNFR is dependent of renal function, endotoxin was injected in nephrectomized mice. The data show that nephrectomy followed by endotoxin injection resulted in a further increase of the levels of both sTNFR. However, the endotoxin induced increase in nephrectomized mice was similar to the situation in normal mice after LPS indicating that the endtoxin induced increase is kidney independent in these mice. To investigate the relative participation of various organs in sTNFR clearance, ¹²⁵I labelled sTNFR-P75 was injected. The data reveal that the majority of the sTNFR is removed from the circulation by the kidneys although indications for involvement of the liver and the lungs were also obtained. Calculation of the parametric clearance revealed that nephrectomy resulted in a 50% reduction of sTNFR-P75 clearance. Furthermore, the data presented strongly suggest that sTNFR release seems to be a continuous process, which is in balance with clearance of the sTNFR by the kidney, although other organs such as the liver and the lungs are involved.     

Occurrence and distribution of free flavonoid aglycones in plants

Flavonoids are widely present in plants as water-soluble glycosides but the lipophilic free aglycones are far less abundant. The 462 flavonoids reported so far to be present in the free state and their plant sources are listed. Evaluation of these data reveals a correlation in most cases between the occurrence of flavonoid aglycones, the presence of secretory structures and the production of other lipophilic plant products. Their accumulation in some plant organs and in certain taxa is discussed. Special attention is given to their occurrence in materials deposited externally on leaves and buds.     

Cav1.2在顺铂诱导C57BL/6J小鼠耳蜗螺旋神经元凋亡中的作用*

目的: 探究顺铂(CDDP)诱导C57BL/6J小鼠耳蜗螺旋神经元(SGNs)凋亡过程中Cav1.2的作用及其可能的机制。方法: 动物实验:选取8周龄雄性C57BL/6J小鼠分为以下两组(10只/组):生理盐水组(Control组)和顺铂给药组(Cisplatin组)。Control组每天腹腔注射生理盐水,Cisplatin组每周期前4 d以3 mg/kg的剂量进行顺铂腹腔注射,后10 d每日注射生理盐水,重复三个周期。给药结束后,听性脑干反应(ABR) 检测小鼠听力阈值变化; 小鼠内眦采血,并断颈取耳蜗,超氧化物歧化酶(SOD)以及丙二醛(MDA)试剂盒检测血清及耳蜗组织的SOD活性和MDA含量;免疫印迹法(Western blot)检测耳蜗组织相关凋亡蛋白表达;苏木精-伊红HE染色观察小鼠耳蜗螺旋神经节形态学变化; TUNEL 染色观察小鼠耳蜗SGNs凋亡情况;免疫荧光观察耳蜗SGNs上Cav1.2的分布和表达。细胞实验:原代培养SGNs,根据CCK8选择顺铂5 μmol/L干预12 h并随机分为:对照组(Control)、溶剂组(DMSO)、Cav1.2阻断剂组(N)、顺铂组(Cisplatin)、顺铂与Cav1.2阻断剂共同孵育组(Cisplatin+N)。Western blot检测Cav1.2蛋白表达;Hoechst33342染色观察各组SGNs凋亡情况,流式细胞术检测各组SGNs凋亡率,Western blot检测相关凋亡蛋白的表达,CA²⁺探针检测细胞内钙离子浓度变化,线粒体膜电位检测试剂盒(JC-1)检测膜电位变化,线粒体超氧化物指示剂(MitoSOX^TM-red)检测线粒体释放ROS情况。结果: 动物实验:与Control组相比,Cisplatin组小鼠听力阈值升高(P<0.01), 血清及耳蜗组织MDA含量、耳蜗组织凋亡蛋白 Cleaved-caspase-3、Bax 蛋白水平和TUNEL阳性率、Cav1.2蛋白表达水平等均明显升高(P<0.05, P<0.01);血清及耳蜗组织SOD活性、耳蜗组织抗凋亡蛋白 Bcl-2 蛋白水平和SGCs密度均明显降低(P<0.05,P<0.01)。细胞实验:与Control组相比,Cisplatin组的Cav1.2表达、细胞凋亡率、Cleaved-caspase-3、Bax蛋白水平、细胞内钙离子浓度以及ROS释放均明显增加(P<0.05,P<0.01);而细胞的Bcl-2蛋白水平和线粒体膜电位则明显降低(P<0.01);Cav1.2阻断剂可部分逆转上述改变(P<0.05)。 结论: 顺铂可能通过上调Cav1.2促进钙内流,进而使线粒体ROS增多,引起SGNs氧化应激损伤从而诱导线粒体途径的细胞凋亡。