[Effects of inner-heating acupuncture on apoptosis of chondrocytes and expression of Caspase-3 and Caspase-9 in rats with knee osteoarthritis].

OBJECTIVE: To investigate the effect of inner-heating acupuncture on apoptosis of chondrocytes and expression of Caspase-3 and Caspase-9 in rats with knee osteoarthritis (KOA). METHODS: A total of 32 rats were divided into a normal group, a model group, a control treatment group and a treatment group by random number grouping method, 8 rats in each one. The rats in the normal group received no intervention. The rats in the remaining three groups adopted modified Videman method to develop KOA model, the ankle joint of left posterior leg was fully extended and fixed with a resin bandage for 6 weeks. After successful modeling, the rats in the model group received no intervention. The rats in the control treatment group were treated with medium-frequency pulse electrotherapy. The rats in the treatment group were treated with inner- heating acupuncture, 30 min each treatment, once a day, five days per week, and totally 3-week treatment was given. After 3 weeks, the damaged cartilage tissue was collected, and HE staining was used to observe the pathological changes of the cartilage tissue of the knee joint. ELISA was used to detect the content of cytochrome-C in the tissue homogenate supernatant. The chondrocytes in damaged cartilage tissue were isolated, flow cytometer was used to detect the changes of apoptosis and mitochondrial membrane potential. The mRNA and protein expression of Caspase-3 and Caspase-9 in chondrocytes were detected by real-time quantitative PCR (qRT-PCR) and Western blot (WB), respectively. RESULTS: Compared with the normal group, the damage of cartilage tissue in the model group was significant, and the expression level of Cyt-C in the homogenate supernatant of damaged cartilage tissue was increased (P<0.01); the chondrocyte apoptosis was increased significantly (P<0.01); the chondrocyte mitochondrial membrane potential was decreased significantly (P<0.01); the mRNA and protein expression of Caspase-3 and Caspase-9 was increased significantly (all P<0.01). Compared with the model group, the cartilage injury in the control treatment group and the treatment group was significantly relieved; the expression level of Cyt-C in the supernatant of damaged cartilage tissue homogenate was decreased (both P<0.01); the chondrocyte apoptosis was significantly reduced (both P<0.01); the chondrocyte mitochondrial membrane potential was increased significantly (both P<0.01). Moreover, the mRNA and protein expression of Caspase-3 and Caspase-9 was significantly reduced (all P<0.01). Compared with the control treatment group, the treatment group was more effective in the treatment of KOA. CONCLUSION: The inner-heating acupuncture could significantly improve the pathological changes of KOA rats, inhibit the apoptosis of chondrocytes, which may be closely related to the suppression of Caspase-3 and Caspase-9 expression.

[Effects of electroacupuncture on the expression of GDNF and Ret in Parkinson's disease model rats].

OBJECTIVE: To explore the mechanism of electroacupuncture therapy on Parkinson's disease (PD). METHODS: Fifty Wistar rats were randomly divided into a normal group, a sham-operation group, a model group, a Fengfu-Taichong group and a Shuanggu Yitong group. PD model was duplicated by microinjection of 6-Hydroxyl-Dopamine into right corpora striata, and by microinjection of normal saline in sham-operation group. Rats in normal group, sham-operation group and model group were not treated. In Fengfu-Taichong group, the rats were treated by electroacupuncture at "Fengfu" (GV 16) and "Taichong" (LR 3) on the basis of the PD model, and by electroacupuncture at "Fengfu" (GV 16), "Taichong" (LR 3), "Guanyuan" (CV 4) and "Zusanli" (ST 36) in Shuanggu Yitong group, once daily for 2 weeks. GDNF and Ret expression were detected by immunohistochemistry and western blotting, respectively. RESULTS: The number of GDNF positive cells and the content of Ret receptor increased significantly in the two electroacupuncture groups compared with those in the other groups (all P < 0.01), and the expression of GDNF increased significantly in Shuanggu Yitong group compared with that in Fengfu-Taichong group (P < 0.01). CONCLUSION: Electroacupuncture can not only increase the expression of GDNF, but also enhance its effect. "Shuanggu Yitong" method is better than simple acupuncture at "Fengfu" (GV 16) and "Taichong" (LR 3) in increasing expression of GDNF.

[Effect of electroacupuncture on morphological changes and apoptosis of substantia nigra cells in Parkinson's disease rats].

OBJECTIVE: To observe the influence of electroacupuncture (EA) of "Fengfu" (GV16) and "Taichong" (LR3) on morphological changes and apoptotic percentage of substantia nigra cells so as to explore its underlying mechanism in relieving Parkinson's disease. METHODS: Forty Wistar rats were randomly divided into normal control, sham-operation (sham), model and EA groups, with 10 rats in each. The Parkinson's disease model was established by micro-injection of 6-hydroxyl-dopamine (6-OHDA) into the right striatum. EA (2 Hz, 1 mA) was applied to "Fengfu"(GV16) and "Taichong" (LR3) for 30 min, once daily for 2 weeks. HE staining, Nissl staining and immunohistochemical staining were conducted for observing the morphological changes of substantia nigra pars compacta (SNc) and striatum, and the tyrosine hydroxylase(TH) immuno-reaction (IR) positive neurons and nerve fibers. The apoptosis was detected by flow cytometry. RESULTS: Compared to the normal control and sham groups, the total number of TH-IR positive neurons and Nissl-stained cells, and OD value of TH-IR positive nerve fibers in the SNc on the 6-OHDA-lesioned side were significantly lower in the model group (P < 0.01). Compared with the model group, the aforementioned 3 indexes of the EA group were increased obviously (P < 0.01). The percentage of apoptosis of SNc on the 6-OHDA-lesioned side was obviously lower in the EA group than in the model group (P < 0.01), but still obviously higher than in the normal group and sham group (P < 0.01). CONCLUSION: Electroacupuncture therapy can significantly increase the number of neurons of substantia nigra and the density of striatum nerve fibers,and reduce the apoptotic percentage of substantia nigra cells in the Parkinsonian rats, which may contribute to its effect in relieving Parkinson's disease.